No full-text available
To read the full-text of this research,
you can request a copy directly from the authors.
Lipid Peroxidation and Antioxidant Enzyme Activities in Infertile Men: Correlation With Semen Parameter
Journal of Clinical Laboratory Analysis
Abstract
The aim of this study was to investigate the oxidative stress status and antioxidant enzyme activities in infertile men's semen and to determine their relationship with spermatozoa characteristics. Four groups of infertile men, normozoospermic (n=9), azoospermic (n=13), oligoasthenozoospermic (n=38), and asthenozoospermic (n=42), were tested for malonaldialdehyde (MDA) concentration and catalase (CAT) and superoxide dismutase (SOD) activities in semen using spectrophotometric methods. We found that MDA levels in semen and SOD activity in seminal plasma (SP) were significantly higher in oligoasthenozoospermic and asthenozoospermic groups compared with normozoospermic group. In contrast, the mean values of CAT activity were not significantly different in all groups. We found positive correlations between semen MDA concentration and SOD activity (P<0.01). MDA levels in sperm pellet and in SP were also positively correlated with mobility grade b (P<0.01), acrosome anomalies (P<0.01), and residual cytoplasmic droplets (P<0.05). In contrast, SOD activity in SP was negatively correlated with sperm concentration and normal morphology (P<0.05). Similarly, CAT activity in SP was negatively correlated with sperm and leukocyte concentrations (P<0.05). In conclusion, our results suggest that the evaluation of oxidative status and antioxidant defenses may be taken as an important tool for diagnosis and treatment of male infertility.
... ROS-induced cell damage occurs due to an unbalanced cell redox, especially when the antioxidant systems cannot compensate for the increased ROS. In the seminal fluid, adequate levels of SOD and CAT play a crucial role in this balance (Abdallah et al., 2009;Khosrowbeygi & Zarghami, 2007;Kobayashi et al., 1991). ...
... Regarding SOD activity, although the two groups that have asthenozoospermia presented slightly lower activity, similarly to other research, we did not find significant differences among the four studied groups (Figure 4a; Hsieh et al., 2002;Khosrowbeygi & Zarghami, 2007;Sanocka et al., 1996;Tavilani et al., 2008;Tkaczuk-Włach et al., 2002). Only Ben Abdallah et al. (2009) described an exception. They found an increased SOD activity in oligoastheno-and asthenozoospermic samples comparing to normozoospermic samples. ...
... They found an increased SOD activity in oligoastheno-and asthenozoospermic samples comparing to normozoospermic samples. Our work evidenced that as SOD levels increase in sperm samples, the CAT, GST, thiol levels and ejaculate volume increase too, and the ejaculate pH decreases (Table 3) concentration (Abdallah et al., 2009;Tavilani et al., 2008) and a negative correlation with sperm concentration and normal morphology (Abdallah et al., 2009) are described in the literature also. ...
Supraphysiological ROS levels can lead to apoptosis, lipid peroxidation, and DNA and protein damage. This pilot study aimed to investigate the sperm oxidative damage in subfertile men, to describe the relationship between the antioxidant system and ROS. Sixty-four semen samples were categorised according to the evaluated routine parameters (WHO, WHO laboratory manual for the examination and processing of human semen, 2010). Results were cross-referenced with the DNA damage [Comet (n = 53) and TUNEL (n = 49) assays], antioxidant enzyme activity [SOD (n = 51), CAT (n = 48) and GST (n = 48)], and content of total thiols (n = 36), lipid hydroperoxide (n = 35) and MDA (n = 31). Compared to pathospermic samples, normozoospermic presented 40%–45% fewer spermatozoa with fragmented DNA, 19% fewer hydroperoxide, and slightly higher total thiols and MDA levels. Asthenozoospermic/asthenoteratozoospermic samples had the lowest GST activity. SOD and CAT showed a similar trend. Our results evidenced significant positive correlations between DNA damage and immotile spermatozoa; SOD and CAT, GST and total thiols; CAT and GST; total thiols and sperm concentration; and MDA levels and head/midpiece abnormalities and hydroperoxides. This work contributes to the existing body of knowledge by showing that the oxidative status correlates with the classic sperm analysis parameters. Oxidative stress and DNA damage evaluation might be a valuable diagnostic and prognostic tool in cases of idiopathic male subfertility.
... In this microenvironment, antioxidant enzymes, such as superoxide dismutase (SOD) and catalase (CAT), can be found. Besides, seminal plasma contains high levels of nonenzymatic antioxidants, such as ascorbate or thiol groups [6]. ...
... Authors concluded that decline in SOD activity might be involved in the abnormal semen quality. On the other hand, Abdallah et al. [6] reported elevated activity of SOD in azoospermic, oligoasthenozoospermic, and asthenozoospermic males compared to normozoospermic ones postulating that SOD expression is upregulated in response to defective spermatogenesis or hormonal deficiency. In other analogous studies, unchanged, elevated, and decreased activities of SOD, CAT, and GPx were found in semen of infertile males compared to the fertile controls [6]. ...
... On the other hand, Abdallah et al. [6] reported elevated activity of SOD in azoospermic, oligoasthenozoospermic, and asthenozoospermic males compared to normozoospermic ones postulating that SOD expression is upregulated in response to defective spermatogenesis or hormonal deficiency. In other analogous studies, unchanged, elevated, and decreased activities of SOD, CAT, and GPx were found in semen of infertile males compared to the fertile controls [6]. The discrepancies between studies may be due to the different study protocols and a result of action of many factors influencing antioxidant enzyme expression and activities. ...
The aim of the study was to evaluate the parameters of oxidative stress and antioxidant defense in relation to the levels of proinflammatory cytokines and chemokines in patients diagnosed with oligozoospermia and asthenozoospermia. Based on the basic parameters of the spermogram, the examined group ( n=243 ) was divided into three groups: oligospermic group (sperm count less than 15 × 10 ⁶ /ml) consisting of 152 men, astenozoospermic group (less than 40% of progressively moving sperm cells) consisting of 142 men, and oligoastenozoospermic group (both criteria met) consisting of 90 men. The control group consisted of 103 males with normal semen profile according to the WHO criteria. Total superoxide dismutase (SOD) activity in seminal plasma and spermatozoa lysate was significantly lower by 12% and 22%, respectively, in males with oligospermia than in the control group. Analogically, Mn-SOD activity in spermatozoa lysate was significantly lower in males with oligospermia, asthenospermia, and oligoasthenospermia by 44%, 32%, and 45%, respectively. By contrast, CuZn-SOD activity in spermatozoa lysate was significantly higher in males with oligospermia by 60%. The activity of glutathione peroxidase (GPx) in seminal plasma was also significantly higher in males with oligospermia and oligoasthenospermia by 56% and 78%, respectively. The level of malondialdehyde (MDA) in seminal plasma was significantly higher in males with asthenospermia than in the control group by 12%. By contrast, the level of MDA in spermatozoa lysate was significantly lower in males with oligospermia, asthenospermia, and oligoasthenospermia by 26%, 20%, and 26%, respectively. The level of interleukin- (IL-) 8 in seminal plasma was significantly higher in males with asthenospermia and oligoasthenospermia by 64% and 67%, respectively. Abnormalities in spermogram, such as oligospermia, asthenospermia, and oligoasthenospermia, may be related to a decreased activity of Mn-SOD in spermatozoa and increased levels of chemokines in seminal plasma.
... Most of the antioxidant buffering capacity is instead active in seminal plasma and includes a number of enzymatic antioxidants such as superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GPx) and catalase (CAT), as well as a variety of non-enzymatic antioxidants such as ascorbic acid (AA) and glutathione (GSH) (Aitken & Roman, 2008;Atig et al., 2012b). Reduced levels of antioxidants (such as SOD, GPx, CAT and AA) are detected in semen of infertile men with varicocele (Abd-Elmoaty et al., 2010) and of men with altered sperm parameters (Ben Abdallah et al., 2009;Atig et al., 2012a,b). ...
... Regarding enzymatic antioxidants, SOD protects sperm from superoxide anions by catalysing the conversion of superoxide into oxygen and oxygen peroxide (H 2 O 2 ), CAT aids in the decomposition of H 2 O 2 into water and oxygen thereby preventing LPO and improving motility . Ben Abdallah et al. (2009) have previously found that the mean values of CAT activity were not significantly different in seminal plasma of oligoasthenozoospermic and asthenozoospermic groups compared with normozoospermic individuals. Moreover, CAT levels were found positively associated with an elevated sperm count (Shiva et al., 2011). ...
... Different studies have investigated the antioxidant ability of seminal plasma in specimens with altered semen parameters, but the results remain controversial, probably for a different inclusion and exclusion criteria used in patient selection, analytical methodologies, sperm anomalies, lifestyle or dietary pattern and the patient's origins. Studies available in literature have selected patients mainly considering sperm parameters (Ben Abdallah et al., 2009;Atig et al., 2012a), whereas the novelty of this research resides in the categorisation of the patients based on their clinical diagnosis: presence of genitourinary infection, varicocele and idiopathic infertility. A similar approach was recently adopted by our group in the study of the relationships between sperm parameters and MDA levels (Collodel et al., 2015). ...
Controls:
In infection and varicocele groups, the seminal MDA levels were significantly increased when compared to controls (p < 0.001), indicating an alteration of oxidative status and a peroxidative damage. In infection and varicocele groups, AA levels were reduced (p < 0.05) vs. control; in the varicocele group, the GSH levels were also decreased (p < 0.05). Significantly higher CAT activity was observed in infection and varicocele groups vs. fertile men (p < 0.001 and p < 0.05 respectively). The GSH/GSSG ratio was significantly decreased in varicocele and idiopathic infertility groups vs. control (p < 0.01). The study of the alteration of a single parameter of oxidative stress or of the antioxidant system may not have a relevant clinical value to estimate male fertilising potential and the background of infertility causes, since complex and multifactorial mechanisms are involved in different pathologies. In our study, each pathology is characterised by a definite pattern of markers such as MDA and enzymatic and non-enzymatic antioxidant compounds. In the different pathologies related to infertility, the identification of the complex of involved parameters could be useful in the diagnosis, prognosis and in the choice of a possible treatment such as specific antioxidant supplements.
... These findings indicate a protective role for antioxidant enzymes of seminal plasma against lipid peroxidation of spermatozoa in normozoospermic samples (Tavilani et al., 2008). Abdallah et al. (2009) found that seminal MDA was significantly higher in oligoasthenozoospermic and asthenozoospermic groups compared with normozoospermic group. Seminal MDA were positively correlated with mobility grade b, acrosome anomalies and residual cytoplasmic droplets. ...
... Seminal MDA were positively correlated with mobility grade b, acrosome anomalies and residual cytoplasmic droplets. In contrast, seminal MDA was negatively correlated with sperm concentrations (Abdallah et al., 2009). ...
... Studies suggested that detection of MDA concentrations in seminal plasma has an indicative value on the diagnosis of male infertility induced by overproduction of reactive oxygen species in male reproductive system (Shang et al., 2004). The evaluation of oxidative status and antioxidant defenses may be taken as an important tool for diagnosis and treatment of male infertility (Abdallah et al., 2009). The measurement of the antioxidative and oxidative agents could serve to evaluate human infertility in those cases where the result of the spematobioscopy appears normal (Gallardo, 2007). ...
Problem statement: Oxidative Stress (OS) has been recognized as one of the most important cause of male infertility. We studied the relation of serum and Semen Malondialdehyde (MDA) and Total Antioxidant Capacity (TAC) with sperm parameters in infertile men with sperm count within the normal range. Approach: The prospective case- control study performed on infertile men presenting to the infertility clinics of Mirzakochak khan hospital, Tehran university of Medical Sciences from June 2007 to June 2009. The samples were collected consecutively and the total of 40 infertile men was enrolled in the study. Also, 40 healthy men were matched as control group. MDA and TAC in serum and seminal plasma were measured and relation between them and semen analysis parameters were evaluated. The MDA was measured as nmol mL-1 and the TAC was expressed as g dL-1. Results: Analysis showed that the amount of semen MDA was statistically different in infertile and healthy control groups. We did not find any significant relation between smoking and sperm parameters in infertile men. The relation between semen MDA and abnormal sperm abnormal morphology (p = 0.003, r = -0.468) and semen TAC and weak sperm motility (p = 0.037, r = -0.359) was significant. Conclusion: Immediate attention should be directed at simplifying and validating the evaluation of reactive oxygen species and OS status so that it can be performed routinely.
... Reactive oxygen species (ROS) are free radicals containing oxygen [59] that can cause sperm cell damage [60]. Oxidants interfere with normal sperm function via membrane lipid peroxidation, having a deleterious effect on the morphological status of sperm cells and, thereby, on male fertilization potential [61]. An important marker of lipid peroxidation is MDA, which to our knowledge has yet to be evaluated in felids. ...
... Semen characteristics and concentrations of MDA in tiger serum and seminal pellets did not differ among the three EE protocols; however, the percentage of primary abnormalities and concentrations of MDA in seminal plasma were significantly higher using the Low voltage EE protocol. In humans, seminal MDA concentrations were positively correlated with abnormal sperm morphology [40,61,62], a finding not observed in this study, although relationships approached significance between MDA and % normal morphology and % secondary abnormalities in the Low group, and for secondary abnormalities in the High voltage group. Thus, the Low voltage protocol might not have fully stimulated the nerves involved in ejaculation, resulting in incomplete or poor-quality ejaculates. ...
Simple Summary
The Bengal tiger (Panthera tigris tigris) is critically endangered, so artificial insemination (AI) is an important conservation tool. Electroejaculation (EE) protocols have been optimized to obtain sufficient amounts of viable sperm for artificial insemination in many felid species; however, less attention has been paid to animal wellbeing during the process. This study examined the effects of three EE protocols (Low, 2–5 volts; Medium, 3–6 volts; and High, 4–7 volts) on semen quality, testicular size, serum creatine kinase (CK, as a proxy for muscle damage), serum testosterone, and cortisol (as a proxy for stress) concentrations, and seminal plasma and sperm pellet malondialdehyde (MDA, as a measure of oxidative stress) measured after each EE series. Primary sperm abnormalities and seminal plasma MDA were higher in the Low compared to Medium and High voltage groups (p < 0.05). Serum CK in the High voltage group increased during the EE series suggesting the potential for muscle damage (p < 0.05). However, no significant differences were observed for serum cortisol, testosterone, or MDA concentrations across voltage groups. Results suggest the Medium voltage protocol produced good quality samples at lower voltages and with no increase in CK compared to the High protocol, which might be better for animal welfare.
Abstract
The Bengal tiger (Panthera tigris tigris) is critically endangered, so assisted reproductive technologies, including artificial insemination, are important conservation tools. For wild and domestic felids, electroejaculation (EE) is the most common semen collection method, with protocols optimized to obtain sufficient amounts of viable sperm for artificial insemination. However, less attention has been paid to ensuring animal wellbeing during the process. This study examined the effects of three EE protocols (Low, 2–5 volts; Medium, 3–6 volts; High, 4–7 volts) on semen quality, testicular size, serum testosterone, creatine kinase (CK), and malondialdehyde (MDA) concentrations, and serum cortisol as a proxy for stress. Blood samples were collected before, during, and after each EE series. Seminal plasma pH, and sperm motility, viability, and morphology were evaluated after each procedure. Seminal plasma and sperm pellet MDA concentrations were also determined. Primary sperm abnormalities and seminal plasma MDA were higher in the Low compared to Medium and High voltage groups (p < 0.05). Serum CK in the High voltage group increased during the EE series (p < 0.05), suggesting the potential for muscle damage. However, no significant changes were observed for serum cortisol, testosterone, or MDA concentrations. Results suggest the Medium voltage protocol produced good quality samples at lower voltages than the High protocol with no negative effect on muscle function, which might be better for animal welfare.
... (37). Many enzymes, as well as transcription factors, have been proposed as potential targets for altered redox status (12). Demonstrate that induced OS is the source of altered steroid hormone receptor expression and distribution, as well as altered gene expression of certain AO enzymes. ...
... Demonstrate that induced OS is the source of altered steroid hormone receptor expression and distribution, as well as altered gene expression of certain AO enzymes. (12). ...
Immunological infertility cause may contribute to up to 8-19% of the male infertility factors, involved in cryptorchidism, testicular trauma, epididymitis, primary testicular failure,idiopathic infertility, infections. As a result, ASA can be present in infertile men who are either primary or secondary infertile. The aim is to look into the relationship between ASA and trace elements in both primary and secondary infertile men. The research was conducted at the ALBasrah Women and Children's Hospital's Infertility Center. This study was included 96 men, divided into two groups infertile and fertile men. Infertile men (66) were subdivided into a primary (33) and secondary infertile men (33).Other (30) fertile men. After (3-5) days of abstinence, seminal fluids were extracted from both of them, and semen fluid analysis was performed on each specimen. Following liquefaction, the concentration, morphology, and motility grades of the sperm were determined (WHO).The mean values of were significantly increased (p<0.05) (CAT, GPx, and TAC)) levels in both primary an d secondary infertile men as compared to fertile men, although the gap was non-significant (P>0.05). (GPx, CAT, and TAC) levels in primary infertile men as compared to secondary infertile men (p>0.05), SOD significant increase (P>0.05) in primary infertile men as compared to fertile men, whereas non-significant increase (P>0.05) in both secondary infertile men as compared to fertile men and primary infertile men as compared to secondary infertile men, When comparing fertile men to primary infertile men and secondary infertile men to fertile men, MDA levels were highly significant increase (p<0.001) in both primary infertile men and secondary infertile men as compared with fertile men,while non-significant increase (P>0.05) in primary infertile men as compared to secondary infertileen. The correlation analysis between ASA and enzymatic antioxidan both immunologically infertile parameters t men were; GPx was positive and significantly correlated; CAT was negative and highly significantly correlated; SOD was non-significant and positive; TAOC was negative and highly significantly correlated;and MDA waspositive and highly significantly correlated.
... The findings of the present study are in accordance with previously published observations of the other authors in studies conducted on human beings (Ben Abdallah et al., 2009;Hashemi et al., 2018). Ben Abdallah et al. (2009) reported that MDA content was elevated in oligozoospermic and asthenozoospermic groups. ...
... The findings of the present study are in accordance with previously published observations of the other authors in studies conducted on human beings (Ben Abdallah et al., 2009;Hashemi et al., 2018). Ben Abdallah et al. (2009) reported that MDA content was elevated in oligozoospermic and asthenozoospermic groups. Murawski et al. (2007) also showed significant and positive correlations between seminal SOD activity and sperm concentration and motility, which are regarded as the important criteria for normal fertilizing ability of the spermatozoa. ...
... LPO produces lipid peroxides that accumulate in spermatozoa, creating a variety of decay end products, such as 4-hydroxynonenal, isoprostanes and malondialdehyde (MDA), which can be measured as indicators of oxidative stress [13]. MDA measurements, the most used method to detect LPO, can be performed either in seminal plasma [14][15][16][17] or sperm cells [9,18]. It was demonstrated that several proinflammatory cytokines, either alone or in the presence of leukocytes, could cause LPO of the spermatozoa plasma membrane to such levels that can affect the sperm fertilising ability [3,9,19]. ...
... Regarding the relationships between MDA levels measured in seminal plasma and sperm parameters, our results are consistent with the literature. In particular, sperm concentration [41,42], motility [14,16,[41][42][43][44], morphology and viability [15], showed significant negative correlations with MDA. Recently, Collodel et al. [17] demonstrated that seminal MDA is a marker of pathologies responsible for sperm motility reduction, such as urogenital infections and inflammatory status. ...
Cytokines are physiological seminal components and their abnormal levels, reported in different pathological conditions, negatively influence the sperm function. We analysed the relationship between interleukin (IL)-6 and IL-33 levels and lipid peroxidation (LPO), measured both in semen and sperm lysate, in 44 human semen samples. The semen analysis was performed following the WHO guidelines. Seminal IL-6 and IL-33 concentrations were assessed by ELISA and LPO was evaluated measuring malondialdehyde (MDA) both in seminal plasma and viable spermatozoa. Two small groups of patients with varicocele and infection were extrapolated from the cases analysed and the variables compared with those of a group of control. IL-33 levels were undetectable in all samples and IL-6 levels were positively correlated with both seminal and sperm MDA concentrations ( p < 0.01) and negatively with sperm parameters ( p < 0.01). Seminal and sperm MDA levels were both negatively correlated with sperm parameters ( p < 0.01). IL-6 and semen MDA showed an exponential positive relationship, whereas MDA values measured in viable spermatozoa were low until IL-6 amount reached a concentration of >30 pg/mL, rising consistently. By comparing the variables in the groups, we confirmed that a high IL-6 concentration in the varicocele and infection groups was concomitant with an increase of seminal MDA levels, but also with MDA measured in viable spermatozoa, which represents the novelty of this study. We identified the IL-6 threshold, beyond which sperm MDA concentration rises concomitantly with the increase of IL-6 concentration. Other studies are needed, considering the increasing number of patients with different pathologies affecting male infertility.
... According literature, very little evidence is found regarding the mechanisms of alcohol-induced oxidative damage to the male reproductive system. Till now, numerous studies reported a negative impact of imbalanced seminal plasma antioxidant or oxidative stress markers on semen parameters in infertile men [20][21][22][23] . Most of these studies compared the correlation between antioxidant activities of blood and seminal plasma on sperm profile in infertile men and confirmed their close relationship with each other [24,25] . ...
... The scavenging potential in gonads and seminal fluid is usually maintained by sufficient levels of SOD and GSHr. SOD has been found in Sertoli cells, germ cells [83] , epididymis and male accessory sex glands, whereas GSH found in testes [20,84] . ...
... The seminal plasma levels of Cd of oligoathenozoospermic group were correlated positively and significantly with MDA levels in seminal plasma .However, there were significant negative correlation between seminal plasma levels of Cd and seminal SOD activity ,sperm count and sperm motility, The explaination is that spermatozoa are particularly susceptible to OS-induced damage, because their plasma membranes contain high concentration of polyunsaturated fatty acids (PUFA) and their cytoplasm contains low concentrations of scavenging enzymes (26) . Lipid hydroperoxides are stable under physiological conditions until they contact transition metals such as cadmium or copper salts. ...
... Both levels of seminal plasma of Cd and MDA in the present study have significant negative correlations with sperm count and motility, these results were inaccordance with Fatma et al. (26) who reported that MDA concentration in semen of all patients was also positively correlated with acrosome abnormality and cytoplasmic retention and the high levels of OS were correlated with impairment of the acrosome reaction owing to protein cross-linking and LP. ...
... In the available studies, MDA levels were predominantly used to estimate the negative effects of oxidative stress in semen (Atig et al., 2012;Ben Abdallah, Dammak, Attia, Hentati, & Ammar-Keskes, 2009;Shiva et al., 2011;Tavilani, Goodarzi, Vaisi-raygani, Salimi, & Hassanzadeh, 2008). In this study, we determined the total oxidant status (TOS) to evaluate the influence of oxidative stress on sperm quality because TOS is related to the level of a single product of LPO and to the levels of the above-mentioned repertoire of products (Erel, 2005). ...
... These results indicate that fertility impairment might be caused by inadequate antioxidant enzyme activities or noncoordination between them. There are studies indicating that poorer semen quality is associated with higher activities of antioxidant enzymes and studies demonstrating that such associations do not exist(Ben Abdallah et al., 2009;Tavilani et al., 2008). The authors who showed elevated activities of these enzymes postulate that their upregulated expression is part of the compensatory defence mechanism against ROS attacks. ...
... We found a direct correlation between the MDA content and the concentration of leukocytes in seminal fluid (r = 0.84; P < 0.05) (Melnyk et al., 2022). MDA level in seminal fluid was negatively correlated with the viability and motility of sperm cells and positively correlated with acrosome abnormalities (Ben Abdallah et al., 2009;Benedetti et al., 2012). This confirms that oxidative damage to lipids of membranes impairs sperm quality (Benedetti et al., 2012). ...
Infertility is a worldwide problem affecting 15% of couples. About 30% cases of male infertility are considered idiopathic associated with factors not limited to testicular dysfunction. Male infertility and rheumatoid arthritis are often associated and patients with rheumatoid arthritis have a lower fertility rate. The exact relationship between rheumatoid arthritis and male infertility has yet to be revealed. The aim of the study is to assess the MDA level and the activity of NO-synthase isoforms and arginase in men with idiopathic infertility and infertility associated with rheumatoid arthritis. Patients were divided into 2 groups: group 1 – 73 patients with idiopathic infertility; group 2 – 68 infertile men with a systemic autoimmune disease (rheumatoid arthritis). The MDA level in blood plasma of patients with idiopathic infertility and infertile men with rheumatoid arthritis was significantly higher than in the control group (fertile men). The cNOS activity in seminal plasma and blood lymphocytes decreased in men with idiopathic infertility and infertility associated with rheumatoid arthritis compared to the control group. The decrease in cNOS activity was accompanied by drastic increase in iNOS activity. The iNOS/cNOS ratio increased in men with idiopathic infertility and men with infertility associated with rheumatoid arthritis. Similar, but more drastic changes in iNOS/cNOS ratio were found in blood lymphocytes. The arginase activity in seminal plasma was decreased in patients with idiopathic infertility compared to the healthy controls. However enzyme activity in men with infertility associated with rheumatoid arthritis had a tendency to increase, but these changes are not significant. Similar changes were observed in blood lymphocytes. It was found that idiopathic infertility is associated with a decrease in the activity of non-oxidative, arginase metabolism and the dominance of NO-synthase, which leads to the hyperproduction of nitric oxide. In men with infertility associated with rheumatoid arthritis an increase in the arginase/NOS ratio in lymphocytes is noted, which is largely explained by the sharp activation of arginase activity in blood lymphocytes. The arginase/NOS balance and iNOS/cNOS ratio may can be used as indicators of oxidative-nitrosative stress in men with idiopathic infertility and infertility associated with rheumatoid arthritis.
... Study by Khosrowbeygi et al. [59] suggested that patient with asthenozoospermia, asthenoteratozoospermia, and oligoasthenoteratozoospermia sperm all had higher levels of PUFA and therefore, were more vulnerable to LPO caused by ROS. Another study has shown similar results, as patients with asthenozoospermia and oligoasthenoteratozoospermia also show higher MDA levels [60]. Furthermore, LPO as measured via MDA levels correlated inversely to in vitro fertilization rates in infertile men [61]. ...
Epidemiological studies regarding male infertility have revealed that more and more infertile men suffer from acute or chronic inflammation of the genitourinary tract, which often occurs without any symptoms. The exact mechanism for unexplained male infertility and its correlation to oxidative stress incidence and/ or the inflammatory reactions within the male genital tract is still controversially unclear till know. Previous studies have shown the presence of cytokines such as tumor necrosis factor-α (TNF-α), a key player in the inflammation process, in the semen of infertile men. However, the mechanism of their effect on human sperm functions is still in need for further under investigations. We expect that, seminal inflammatory cytokine (TNF-α) and its expression regulator, testicular ADAM 17, will be varied, when seminal OS is established, that in turn, will vary cellular integrity and energy status. As conventional treatment of male infertility such as mineral and vitamins supplementation has poor success rates and leaves a lot to be desired, natural antioxidant ingredients are greatly suggested, nowadays, to ameliorate oxidative stress and the inflammatory detrimental effects accompanied to idiopathic male infertility. Thus, the empirical investigations of naturally occurring antioxidant modulators with prospective curative effect may support their administration as safe potent natural remedies against IMI. The role of antioxidants in the treatment of unexplained male infertility correlation to ROS and/or the inflammatory cytokines are discussed in this review. by routine semen analysis is considered normal. Men with idiopathic infertility generally present with significantly higher seminal reactive oxygen species (ROS) levels and lower antioxidant potential than healthy fertile controls [3]. In addition, high ROS levels have been detected in the semen samples of 25% to 40% of infertile men [4]. Evidence now suggests that ROS-mediated damage to sperm is a significant contributing pathology in 30-80% of cases [5].
DOI: https://doi.org/10.4172/2469-5653.1000107
... Our study showed a significant elevation in the seminal MDA levels in infertile groups. Therefore, lipid peroxidation degradation of sperm membrane integrity could held responsible for abnormal sperm motility, concentration and morphology [26]. Some studies appeared the negative significant correlation was noticed between lipid peroxidation with sperm concentration, and normal morphology between control and infertile men [27] [28]. ...
The study was carried out between January 2014 and October 2014 including one hundred and fifty five infertile men who attended the Fertility center in AL-Sadr Medical City. The Objective of this study was to study of oxidative stress, sperm chromatin abnormality and semen quality including (semen volume, Liquefaction time, sperm concentration, motility, and normal morphology. The results of the study showed that there was a significant increase (p<0.05) in the semen volume, sperm concentration, progressive motility and normal morphology for unexplained compared to five abnormal semen groups. Also it was a significantly decrease (p<0.05) in liquefaction time, leukocyte concentration, MDA concentration and sperm chromatin abnormality for unexplained compared with abnormal semen groups. This study supports the conclusion that oxidative stress in seminal plasma is the main causes of sperm chromatin defect with defect of semen parameters and related with male infertility.
... These findings were similar to the finding of Hsieh et al., Kobayashi et al., and Athayde et al. [10,11] Similarly, an elevated level of MDA was also noted in oligoasthenoteratozoospermia in a study conducted by Fraczek et al., 2001. [12] A prospective study demonstrated that men with high levels of ROS generation had less chance of initiating a pregnancy compared with low ROS. ...
... Hyperglycaemia induces oxidative stress resulting in diabetic complications (Oyenihi et al. 2015). Increased oxidative stress can lead to the aldehydic metabolism of lipid peroxidation, generating MDA molecules (Gaweł et al. 2004), elevated levels of which in the seminal plasma cause sperm might damage after ejaculation (Abdallah et al. 2009). Antioxidant supplements have been shown to correct MDA levels in the seminal vesicle plasma and tissue of DM animals (Tsounapi et al. 2017), which is similar to our findings. ...
Context
Although Dolichandrone serrulata (Wall. ex DC.) Seem (Bignoniaceae) flower (DSF) improves hyperglycaemia, testicular damage and sperm quality in type 2 diabetes mellitus (T2DM) animals, its effects on the seminal vesicles, secreting seminal plasma, are unknown.
Objective
This study reports the protective effects of DSF on seminal dysfunction in T2DM rats.
Materials and methods
Male Sprague-Dawley rats were divided into four groups (control, T2DM, T2DM + DSF200 and T2DM + DSF600; 10 animals/group). The control group was fed a low-fat diet for 14 days prior to single saline injection, whereas T2DM group was given a high-fat diet and injected with streptozocin (50 mg/kg body weight). The T2DM-induced rats were fed DSF orogastrically (200 and 600 mg/kg body weight) for 28 consecutive days. At the end of the experiment, biochemical components, malondialdehyde (MDA), histology and protein expression in seminal lysate were evaluated.
Results
DSF increased the levels of serum phosphorus (13.66 ± 0.59 mg/dL), ALP (11.85 ± 0.99 U/L), GOT (3938.23 ± 251.41 U/L) and GPT (34.16 ± 4.93), decreased MDA levels in seminal tissue, and elevated the serum testosterone in the T2DM rats. Treatment with DSF ameliorated histological damage, significantly increased seminal 44 and 31 kDa TyrPho protein expression, and decreased that of caspase 3 and 9.
Conclusions
DSF extract was able to mitigate seminal dysfunction in T2DM rats via improvements of tyrosine phosphorylation, testosterone level and biochemical substances, as well as reductions of caspase proteins. DSF may be developed as an alternative medicine in treating of T2DM male subfertility and progressive complications.
... It has also been reported that seminal plasma acts as a natural source of antioxidant, which protects the sperm cell from ROS. 76,86 Similar findings were also reported in the case of ram semen cryopreservation with tris extender containing additional seminal plasma of other species. 75 GSH-Px activity was significantly decreased in frozenthawed semen as compared with fresh semen. ...
The present study was conducted to observe the effects of removal of seminal plasma of Pantja buck semen and supplementation of bovine seminal plasma (BSP) in the extender before cryopreservation. In a preliminary experiment, different levels of BSP were supplemented (1, 3, 5, 7, and 9% v/v) in egg yolk (7.5% egg yolk)-tris (EYT) extender and used for cryopreservation of Pantja buck semen. Results in terms of motility, viability, plasma membrane integrity, acrosome integrity, and lipid peroxidation showed that 5% BSP was suitable for maintaining Pantja buck semen quality during cryopreservation. In the final experiment, pooled semen from four Pantja bucks was split into three aliquots (I, II, and III). Aliquot I was directly diluted in EYT extender and grouped as the control (C); aliquot II and III were washed separately with TALP solution and diluted as D1 (Washed semen with EYT extender) and D2 (Washed semen with EYT extender containing 5% BSP), respectively. Seminal attributes (sperm individual motility, viability, plasma membrane integrity, acrosome integrity, and total morphological abnormalities) were assessed at the postdilution, postequilibration, and post-thawing stages. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA) concentration, and glutathione peroxidase (GSH-Px) activity were measured at post-thaw. Washed semen significantly improved (p < 0.05) seminal parameters at post-thaw compared with unwashed semen (control). A significant difference (p < 0.05) was observed in seminal attributes between freezing stages and between dilution groups. Significantly higher (p < 0.05) post-thaw sperm motility, viability, plasma membrane integrity, acrosome integrity, and GSH-Px activity, and significantly lower (p < 0.05) MDA concentration and extracellular release of enzymes (ALT, AST) were observed in group D2 compared with control and D1. The results of the present study demonstrated that cryopreservation of washed Pantja buck semen diluted with 5% BSP-supplemented EYT extender can improve post-thaw semen quality.
... Moreover, it may increase the number of leukocytes in semen and lead to hormone fluctuations (e.g., testosterone), causing disorders in the functioning of the male reproductive system and significantly reducing the ability to fertilize (3,(8)(9)(10). Thus, assessing the level of oxidative stress in the body should be included in male infertility diagnosis, and may be a useful tool in determining the ability to fertilize sperm (11). ...
The influence of fruit and vegetable consumption on semen quality by reducing oxidative stress is inconsistent. Thus, the association between the consumption of these products, antioxidant status, and semen quality was investigated in 90 men aged 18–40. The consumption of fruit and vegetables was collected using the 3-day food record method. Antioxidant status: total antioxidant capacity in semen (TAC-s) and blood (TAC-b), blood superoxide dismutase (SOD-b), glutathione reductase (GR-b), glutathione peroxidase (GPx-b), catalase (CAT-b) activity, and malondialdehyde concentration in blood (MDA-b) were measured. Sperm concentration, leukocytes in the ejaculate, vitality, motility, and sperm morphology were examined using computer-aided semen analysis (CASA). The consumption of fruit and vegetables was positively correlated with sperm concentration, vitality, motility, TAC-s, TAC-b, and SOD-b activity. The TAC-s and TAC-b were positively related to motility, TAC-s was inversely correlated with sperm tail defects. The SOD-b activity was positively correlated with vitality, motility, sperm morphology, and inversely with sperm tail defects and leukocytes in the ejaculate. Compared to the men in the first quartile of fruit and vegetable consumption (<318 g/day), those in the highest quartile (>734 g/day) had the highest sperm concentration, vitality, motility, TAC-s, TAC-b, GPx-b activity, and the lowest MDA-b concentration (based on multivariate regression models). A high consumption of fruit and vegetables may positively influence selected sperm quality parameters by improving the antioxidant status of semen and blood.
... Another study reported that the seminal plasma of infertile men presented higher SOD activity, while the CAT activity was similar to that of seminal plasma of fertile men [102]. Yet, another study described that SOD and CAT activities were correlated with sperm concentration [103], both being enhanced in subfertile/infertile men. This highlights contradictory results between the studies published on the subject. ...
Nowadays, infertility is classified as a disease of the reproductive system. Although it does not compromise the life of the individual, it can have detrimental effects on the physiological and psychological health of the couple. Male fertility evaluation is mainly focused on the analysis of sperm parameters. However, the ejaculated fluid is also composed of seminal plasma, and the study of this fluid can provide crucial information to help in the assessment of male fertility status. Total antioxidant capacity of the seminal plasma has been positively correlated with the fertility of men. Moreover, evidence highlights to a similar importance as that of female reproductive tract fluid antioxidant capabilities and female fertility. Herein, we describe the functions of seminal plasma and female reproductive tract fluids, as well as their main antioxidant components and their relationships with fertility outcomes. Additionally, this review contains the most up to date information regarding the mechanisms of the interaction between the male and the female reproductive fluids and the importance of proper antioxidant capacity for fertilization.
... The presence of antioxidant enzymes GSH-Px, SOD, and CAT have been reported to counteract the effects of ROS and LPO to ensure sperm protection in both seminal plasma and spermatozoa of poultry such as chickens, turkeys, and guinea fowls and in mammals (Khan 2011;Michael et al. 2007;Surai et al. 1998b;Zhang et al. 2017). Studies of fresh chicken and goose semen revealed that antioxidant enzymes CAT and SOD increase their activities in proportion to the increase in ROS and LPO, neutralizing their effects and maintaining sperm quality (Ben Abdallah et al. 2009;Partyka et al. 2012b). Also, the activity of SOD was reported to increase with increases in ROS and LPO in frozen semen of ducks and chickens (Partyka et al. 2013;Surai et al. 2000). ...
Plasma membrane lipids are the key factor in the ability of chicken sperm to be frozen. They ensure fluidity and flexibility of the sperm membrane for effective viability and motility during in vitro storage. The objective of this study was to compare the lipid profiles of different native and commercial chicken breeds: native Thai (Pradu Hang Dam) roosters and commercial (Rhode Island Red) roosters, with respect to their frozen sperm quality. In addition, lipid peroxidation and antioxidant enzymes superoxidase dismutase (SOD) and catalase (CAT) were also examined. Semen was collected from 12 roosters of each breed. For fresh semen, parameters assessed include semen volume, pH, sperm concentration, sperm motility, and viability, while for frozen semen, the parameters assessed were sperm motility and viability. Moreover, other parameters assessed included malondialdehyde (MDA) concentration, activities of SOD and CAT, and fatty acid profile. We found that sperm viability and motility of frozen semen were higher in the commercial breed than in the native breed (P < 0.05). The commercial chicken breed had higher MDA concentration than the native breed (P < 0.05), but antioxidant enzymes remained unchanged in both. Levels of arachidonic acid (AA; C20:4n-6) and docosahexaenoic acid (DHA; C22:6n-3) were significantly higher (P < 0.05) in the commercial than in the native breed; however, n-6 to n-3 ratios were not different. In conclusion, our study found that lipid profiles have an influence on frozen sperm viability and motility between the breeds. Polyunsaturated fatty acids, particularly AA and DHA, are beneficial to sperm quality.
... These findings were similar to the finding of Hsieh et al., Kobayashi et al., and Athayde et al. [10,11] Similarly, an elevated level of MDA was also noted in oligoasthenoteratozoospermia in a study conducted by Fraczek et al., 2001. [12] A prospective study demonstrated that men with high levels of ROS generation had less chance of initiating a pregnancy compared with low ROS. ...
... Our data were consistent with previous studies that found no significant change in catalase levels between asthenozoospermic, oligozoospermic and normozoospermic groups (Tomar, et al. 2017. Ben Abdallah, 2009). Still, several other investigations have found elevated rates of catalase activity in normozoospermic compared to the abnormal groups [ Shamsi,2009, Vatannejad, 2017]. Yet, a recent study found that the activity of catalase was decreased in seminal plasma of ejaculate from males with normal sperm motility compared with semen from asth ...
... This imbalance made the ability of spermatozoa antioxidants in scavenging the oxidants become limited, and increased the rate of oxidative stress. The present results of MDA were in agreement with the results of the other authors (Tavilani et al., 2008;Ben Abdallah et al., 2009;Huang et al., 2000). ...
Spermatogenesis is the most important stage in male fertility. In the present study, many factors affecting spermatogenesis were evaluated in seminal plasma and homogenates of sperms from fertile and infertile males. The levels of some heavy metals, malondialdehyde (MDA) and reduced glutathione (GSH) as well as superoxide dismutase (SOD) activity were measured in both seminal plasma and homogenate supernatant of sperm from fertile and infertile males. Two hundred and sixteen human semen samples were collected and divided into five groups: group I (GI) included 70 semen samples as the control, and four infertile groups (GII-GV) according to WHO criteria. The levels of Fe2+, Cd2+ and MDA in seminal plasma and sperm homogenate supernatant of infertile groups were highly and significantly increased compared to their levels in control. However, GSH level and SOD activity in seminal plasma and sperm homogenate supernatant of infertile men were highly and significantly decreased compared to the control. In conclusion, the levels of heavy metals and oxidative stress were associated with human male spermatogenesis dysfunction and might be useful tools in predicting sperm density.
... Thus, seminal plasma MDA or sperm MDA can be regarded as useful dual biomarkers of oxidative damage to sperm and can be used effectively for an analysis of infertility or risk assessment. The literature has demonstrated that the MDA level in seminal plasma negatively correlates with sperm motility, sperm concentration, and sperm viability (Atig et al., 2012;Das et al., 2009;Hsieh et al., 2006); the MDA level has also been reported to positively correlate with acrosome anomalies (Ben Abdallah et al., 2009). Consequently, the elevation of MDA in seminal plasma indicates that sperm suffered oxidative damage and that some functions of sperm might have been impaired. ...
Background:
Current available evidence regarding the detrimental effects of low-level ambient air pollution on conventional semen parameters is inconclusive. In nonreproductive systems, air pollutant exposure has been demonstrated to induce oxidative stress (OS), which is a crucial mechanism that mediates sperm damage and male infertility. Thus, it may be essential to investigate the effects of air pollution on sperm quality in terms of the perspectives of OS and relative molecular damage.
Objectives:
We assessed the associations of major air pollutant exposure to oxidative stress-mediated alterations in semen, including seminal plasma malondialdehyde (MDA), sperm mtDNA copy number, and integrity.
Methods:
The present study used data gathered from 516 young men participating in the Male Reproductive Health in Chongqing College student (MARCHS) cohort study during the follow-up stage in 2014 (n = 427 on the old campus, which is located in an urban area and has worse air quality, and n = 89 on the new campus, which is not urban and has better air quality). Data regarding major air pollutant exposure during 0-90, 0-9, 10-14 and 70-90 days before each semen examination (corresponding to the entire and three key periods of sperm development, respectively) were collected. The Mann-Whitney U nonparametric test was employed to compare distributions of major air pollutants and to explore differences in MDA, mtDNA copy number, and mtDNA integrity between the two campuses. A linear regression model was used as multivariable analysis to investigate associations of major air pollutant exposure with these biomarkers of oxidative damage to sperm and to adjust for potential confounders.
Results:
During all four key periods of sperm development, compared with college students on the new campus, college students on the old campus were exposed to higher levels of PM10, PM2.5, NO2, and CO, and had higher air quality index (AQI) values, indicating that these participants suffered from worse air quality. The levels of seminal plasma MDA in college students on the old campus were higher than those for the new campus (2.0 nmol/ml; 0.7, 3.6 vs. 1.6 nmol/ml; 0.4, 3.4, p < 0.001) (medians with 5th and 95th percentiles). There were no significant differences in sperm mtDNA copy number and mtDNA integrity between the two campuses. Furthermore, daily average PM10 exposure during 0-90 days before semen ejaculation was found to be significantly and positively associated with seminal plasma MDA level (10.4; 95% CI, 4.4, 16.4) (percentage change per 10-unit increase in air pollutant concentration; same meanings for the results below); daily average SO2 exposure for 70-90 days and NO2 exposure for 0-9 days prior to sampling were also positively associated with MDA level (74.7; 95% CI, 32.1, 119 and 11.9; 95% CI, 4.8, 19.0, respectively). AQI for 0-90 days and 70-90 days prior to sampling positively correlated with seminal plasma MDA concentrations (11.4; 95% CI, 4.7, 18.1 and 12.2; 95% CI, 5.3, 19.1, respectively). Additionally, daily average SO2 exposures for 10-14 and 0-9 days prior to sampling were negatively associated with sperm mtDNA copy number and mtDNA integrity, respectively (-9.0; 95% CI, -16.4, -1.6 and -38.3; 95% CI, -64.1, -11.8, respectively). However, only the correlations between SO2 exposure and AQI value for 70-90 days prior to sampling and MDA levels remained significant after multiplicity adjustment.
Conclusions:
The results indicate that bad air quality, especially SO2 exposure during certain periods of sperm development, might be correlated with oxidative damage to sperm. These findings can deepen the understanding of the potential impacts of air pollution on sperm quality.
... However, the high concentrations of Rutin (2.0 mM) did not result in any significant ameliorative effect on the physiological characteristics of sperm. It might be supposed that the new lipid environment was caused by the high concentration of Rutin, and lipid peroxidation might still represent a cause of sperm damages and decreased sperm quality.Sperm is highly sensitive to ROS and lipid oxidation, especially in frozen-thawed sperm (Allamaneni, Naughton, Sharma,Thomas, & Agarwal, 2004;Ben, Dammak, Attia, Hentati, & Ammar-Keskes, 2009;Liu, Wang, et al., 2018;Wang, Zhang, Ikemoto, Anderson, & Loughlin, 1997). To analyze the mechanism of the ameliorative effect of Rutin on post-thaw sperm, we examined the amount of ROS production and OS in frozen-thawed spermatozoa. ...
This study was aimed to investigate whether and how Rutin protects boar sperm against cryoinjury during cryopreservation. Five concentrations of Rutin with 0.2, 0.4, 0.6, 1.0, and 2.0 mM were added to the freezing extender of boar sperm, respectively, and the effects on quality and function of boar sperm after freezing‐thawing were assessed. The results showed that the sperm motility, mitochondrial activity, plasma membrane integrity, and acrosomal integrity were significantly improved in 0.4 mM and 0.6 mM Rutin groups (p < .05). Compared with ganoderma lucidum polysaccharide (GLP) or Tanshinone IIA, Rutin exhibited higher rates of mitochondrial activity and acrosome integrity (p < .05). Mechanistically, the addition of Rutin at the concentration of 0.6, 0.8, and 1.0 mM significantly attenuated ROS accumulation and MDA production by improving antioxidant enzymatic activity, including SOD, CAT, and GSH‐Px (p < .05). Functionally, a higher penetration rate and the increased total efficiency of fertilization were observed in the 0.4, 0.6, and 1.0 mM Rutin groups than in the control group (p < .05). Moreover, the addition of Rutin (0.6 mM) significantly induced an increase in both the cleavage and blastocyst rates (p < .05). In summary, supplementation with Rutin in cryopreservation medium protects boar sperm against ROS attack by enhancing the antioxidative defense.
... The human body produces endogenous antioxidants in an effort to prevent the damage caused by ROS [91,92], but this response is not always adequate, resulting in OS. Several studies have shown that exogenous antioxi-dants have the capacity to counteract oxidative damage or OS, improving both sperm motility and DNA integrity for infertile men with OS (Table 3) [87][88][89][90][91][93][94][95][96][97][98][99][100][101][102][103][104][105][106][107][108][109][110][111]. Indeed, many oral formulations of antioxidants are readily available in the market and are commonly used to treat men with infertility. ...
Despite advances in the field of male reproductive health, idiopathic male infertility, in which a man has altered semen characteristics without an identifiable cause and there is no female factor infertility, remains a challenging condition to diag- nose and manage. Increasing evidence suggests that oxidative stress (OS) plays an independent role in the etiology of male infertility, with 30% to 80% of infertile men having elevated seminal reactive oxygen species levels. OS can negatively affect fertility via a number of pathways, including interference with capacitation and possible damage to sperm membrane and DNA, which may impair the sperm’s potential to fertilize an egg and develop into a healthy embryo. Adequate evaluation of male reproductive potential should therefore include an assessment of sperm OS. We propose the term Male Oxidative Stress Infertility, or MOSI, as a novel descriptor for infertile men with abnormal semen characteristics and OS, including many patients who were previously classified as having idiopathic male infertility. Oxidation-reduction potential (ORP) can be a useful clinical biomarker for the classification of MOSI, as it takes into account the levels of both oxidants and reductants (antioxidants). Current treatment protocols for OS, including the use of antioxidants, are not evidence-based and have the potential for complications and increased healthcare-related expenditures. Utilizing an easy, reproducible, and cost-effective test to measure ORP may provide a more targeted, reliable approach for administering antioxidant therapy while minimizing the risk of antioxidant overdose. With the increasing awareness and understanding of MOSI as a distinct male infertility diag- nosis, future research endeavors can facilitate the development of evidence-based treatments that target its underlying cause.
... Because of the small amount of cytoplasm in sperm, which is removed during spermiogenesis, seminal plasma has an important role in ROS defense through several nonenzymatic and enzymatic antioxidants. Nonetheless, extender dilution before vitrification decreases the antioxidants constituents so that the resulting sperm is vulnerable to OS [27]. The proper level of ROS is necessary for sperm function. ...
Background
Vitrification is the main technique in the assisted reproductive technique (ART) labs. Sperm vitrification exposes sperm to damage. The aim of the present study was to evaluate the in vitro effect of Ceratonia siliqua (C. siliqua) application on sperm parameters and chromatin quality in normozoospermic aged men. Semen samples ( n = 40) were collected from normozoospermic men over 45 years old. Each specimen was divided into four aliquots to form the subsequent groups: fresh (group I), vitrification without treatment (group II), vitrification with the medium supplemented by 20 μg/ml C. siliqua (group III), and vitrification with the thawing medium supplemented by 20 μg/ml C. siliqua (group IV). Sperm progressive motility, normal morphology and viability were assessed. Also, sperm chromatin quality was evaluated by aniline blue (AB), toluidine blue (TB), and sperm chromatin dispersion (SCD) staining.
Results
Vitrification caused a significant decrease in sperm progressive motility, normal morphology and viability as well as chromatin quality compared to fresh samples ( p < 0.05). Supplementation of vitrification/thawing medium with C. siliqua significantly improved sperm progressive motility, normal morphology, viability, and chromatin quality compared to vitrification without any supplementation ( p < 0.05).
Conclusions
The study showed that C. siliqua can improve the detrimental effect of vitrification on sperm parameters and chromatin quality of normozoospermic aged men.
... The human body produces endogenous antioxidants in an effort to prevent the damage caused by ROS [91,92], but this response is not always adequate, resulting in OS. Several studies have shown that exogenous antioxi-dants have the capacity to counteract oxidative damage or OS, improving both sperm motility and DNA integrity for infertile men with OS (Table 3) [87][88][89][90][91][93][94][95][96][97][98][99][100][101][102][103][104][105][106][107][108][109][110][111]. Indeed, many oral formulations of antioxidants are readily available in the market and are commonly used to treat men with infertility. ...
Despite advances in the field of male reproductive health, idiopathic male infertility, in which a man has altered semen
characteristics without an identifiable cause and there is no female factor infertility, remains a challenging condition to diagnose
and manage. Increasing evidence suggests that oxidative stress (OS) plays an independent role in the etiology of male
infertility, with 30% to 80% of infertile men having elevated seminal reactive oxygen species levels. OS can negatively affect
fertility via a number of pathways, including interference with capacitation and possible damage to sperm membrane and
DNA, which may impair the sperm’s potential to fertilize an egg and develop into a healthy embryo. Adequate evaluation of
male reproductive potential should therefore include an assessment of sperm OS. We propose the term Male Oxidative Stress
Infertility, or MOSI, as a novel descriptor for infertile men with abnormal semen characteristics and OS, including many
patients who were previously classified as having idiopathic male infertility. Oxidation-reduction potential (ORP) can be a
useful clinical biomarker for the classification of MOSI, as it takes into account the levels of both oxidants and reductants
(antioxidants). Current treatment protocols for OS, including the use of antioxidants, are not evidence-based and have the
potential for complications and increased healthcare-related expenditures. Utilizing an easy, reproducible, and cost-effective test to measure ORP may provide a more targeted, reliable approach for administering antioxidant therapy while minimizing
the risk of antioxidant overdose. With the increasing awareness and understanding of MOSI as a distinct male infertility diagnosis,
future research endeavors can facilitate the development of evidence-based treatments that target its underlying cause.
... Because of the low content of cytoplasm in spermatozoa, which is eliminated during the final stages of spermatogenesis, seminal plasma provides the major defense against ROS through some enzymatic and nonenzymatic antioxidants (13). Nevertheless, dilution of the extender before cryopreservation diminishes the concentration of these constituents present in seminal plasma, thereby leaving spermatozoa vulnerable to OS. ...
Objective
To investigate the potential effects of TAT-PRDX2 protein supplementation to the cryopreservation medium on post-thaw sperm quality and function.
Design
In vitro prospective study.
Setting
Medical university hospital.
Patient(s)
Fifty normozoospermic, 50 asthenozoospermic, and 50 oligoasthenozoospermic men undergoing semen analysis for couple infertility.
Intervention(s)
Each semen sample was divided into three aliquots: fresh, cryopreserved control (without additive), and cryopreserved with TAT-PRDX2 protein.
Main Outcome Measure(s)
Sperm motility, viability, mitochondrial potential, and DNA damage as well as reactive oxygen species (ROS) levels and lipid peroxidation were analyzed. Acrosome reaction and zona-free hamster oocyte penetration tests were performed to assess the fertilization ability of cryopreserved spermatozoa.
Result(s)
In normozoospermic and asthenozoospermic groups, the addition of 150 μg/mL TAT-PRDX2 significantly reduced intracellular ROS and malondialdehyde levels and enhanced post-thaw sperm motility and viability when compared with the cryopreserved control of the respective groups but did not produce any significant protective effect in the oligoasthenozoospermic group. Mitochondrial potential was significantly increased, whereas DNA fragmentation was significantly decreased, after TAT-PRDX2 supplementation only in the asthenozoospermic group when compared with the cryopreserved control. Although the penetration rate and the penetration index were not markedly improved, TAT-PRDX2 supplementation obviously reduced spontaneous acrosome reaction and increased calcium ionophore–induced acrosome reaction in the normozoospermic and asthenozoospermic groups.
Conclusion(s)
TAT-PRDX2 protein effectively exerted cryoprotective effects on spermatozoa by reducing intracellular ROS level and thereby improved post-thaw sperm quality and function, especially for asthenozoospermic samples. TAT-PRDX2 protein is a promising additive for developing a new and highly efficient semen cryoprotectant.
... At this point, since supra-therapeutic doses were ineffective to produce oxidative stress, a very high dose of CGV given for a long period of time may be required. There have been positive correlations between total activities of reduced glutathione, catalase, superoxide dismutase and glutathione peroxidase with total content of MDA in seminal plasma from normozoospermic samples [56]. However, antioxidant enzyme activities were modulated by CGV. ...
Background:
The misconception about dietary supplements being safe has led many into the in-patient wards. Cellgevity® (CGV) is a Max International premiere antioxidant supplement formula used by a large population. This study evaluated the effects of therapeutic and supra-therapeutic doses of CGV on reproductive function and biochemical indices in Wistar rats.
Methods:
Seventy-two Wistar rats weighing 130 ± 15.8 g were grouped into two categories (male or female) of six rats per group. Control group received distilled water (10 ml/kg). Others received therapeutic (14.3 mg/kg or 28.6 mg/kg) and supra-therapeutic CGV doses (1000, 2000 or 3000 mg/kg) body weight per oral respectively.
Results:
After 60 days, supra-therapeutic doses of CGV reduced sperm motility (p < 0.05) by 31.8%, 31.3% and 34.5% respectively and increased (p < 0.05) abnormality in sperms by 200%, 241% and 141.3% respectively. CGV altered male (luteinizing, follicle stimulating hormones and testosterone) and female reproductive hormones (luteinizing, follicle stimulating hormones estrogen and progesterone) respectively. Therapeutic doses of CGV elevated reduced glutathione, superoxide dismutase, catalase and glutathione S-transferase, although, this was exceeded by supra-therapeutic doses and more in females than male rats. Supra-therapeutic dose (3000 mg/kg CGV) decreased body weight in both male and female rats by 50% (F(1.5, 30) = 1.2, p = 0.041) and 62.7% (F(2.1, 30) = 0.38, p = 0.038) respectively in treated rats. Supratherapeutic (3000 mg/kg) dose of CGV increased (p < 0.05) creatinine level by 99.1% while serum total protein was reduced (p < 0.05) by 60.1% (2000 mg/kg) and 57.2% (3000 mg/kg) respectively in male animals. In Female rats, supra-therapeutic doses of CGV elevated creatinine levels by 72.2% (1000 mg/kg), 60.2% (2000 mg/kg) and 124.8% (3000 mg/kg) respectively and 3000 mg/kg produces elevated serum low density lipoprotein by 34.6% in treated rats. Serum cholesterol, triglycerides, albumin, alkaline phosphatase were unaltered by CGV dosing. Histology shows seminiferous tubules with reduced spermatogenic cells. Also, female rat kidney revealed acute tubular necrosis at highest dose used in this study.
Conclusion:
Overall, these data suggest that pro-oxidant potential of the supra-therapeutic CGV doses is evident. Hence, it is necessary that its administration be done with caution using appropriate doses.
... We also found a significantly lower CAT activity level in males with normal sperm motility compared to asthenozoospermic males. Other researchers including Khosrowbeygi, Zarghami [64], Tavilani et al. [65], Ben Abdallah et al. [66], and Shiva et al. [67] also found positive correlations between sperm motility and CAT activity. Interrogation of our data found a significant negative correlation between CAT activity in seminal plasma and the volume of ejaculate, while Zini et al. [62,63] did not show these relationships. ...
We analyzed cobalt (Co), chromium (Cr), and lead (Pb) concentrations in human semen and catalase CAT activity in seminal plasma and the effects of their relations on the sperm quality. We obtained semen samples from men (n = 168) undergoing routine infertility evaluation. Studies included two groups based on the ejaculate parameters: I (n = 39; normal ejaculate; normozoospermia); II (n = 129; pathological spermiogram). We examined relationships and differences between Co, Cr, and Pb concentrations in seminal plasma, CAT activity, and semen parameters. We did not establish differences in Co, Cr, and Pb concentrations and CAT activity from men between normozoospermic and those with pathological spermiogram. We found a significantly lower Co concentration and CAT activity in males with normal sperm motility than in asthenozoospermic males. We found significantly lower Co and a higher Pb concentration in males with normal morphology of spermatozoa than in teratozoospermic males. We found a significantly higher Pb concentration in the individuals with consumption of alcohol than in those without consumption. There were significant correlations between Co and Pb concentrations, sperm progressive motility (A + B, i.e., fast and slow progressive motility; Co-negatively; Pb-positively), and normal morphology of spermatozoa (Co-negatively; Pb-positively). We found a significant negative correlation between Cr concentration and slow progressive motility, and between CAT activity and volume of ejaculate. Co, Cr, and Pb levels and CAT activity were related to sperm characteristics and male fertility. The impact of alcohol may be manifested by a disturbance in Pb equilibrium in the body. Co and Pb influence progressive motility and normal morphology of human spermatozoa. Thus, Co and Pb levels in semen may be a useful diagnostic in male infertility. Most of the results of this study are in contrast to expectations. Namely, Pb is a toxic element and its harmful effects (poor semen quality) may be expected already at relatively low level of Pb exposure and are particularly visible with increasing of Pb. Co and Cr(III) are essential elements and harmful effects may be expected at their deficiency and/or overexposure.
... It is known that alterations in intracellular ROS could be associated with male idiopathic infertility [16]. These results are in agreement with data of our previous publications [18] and some other studies [4,27] also demonstrating that the increased malone-dialdehydelevel(evaluatedasTBARS)insemenwassignificantlyhigherinoligoasthenozoospermic and asthenozoospermic groups compared with normozoospermic group. ...
The level of the products of lipid peroxidation measured as the thiobarbituric acid reactive substances (TBARS) and activity of enzymes of glutathione antioxidant protection system in spermatozoa were studied. It was shown that mean levels of TBARS in sperm cells of infertile men with oligozoo-, astenozoo-, oligoastenozoo- and leucocytospermia were 1.6-, 2.0-, 2.0- and 2.7-fold higher, respectively, compared with such levels in the fertile men. The glutathione peroxidase activity was decreased 2.2-fold in patients with oligozoospermia, 2.8-fold in patients with astenozoo- and oligoastenozoospermia and 3.8-fold in patients with leucocytospermia. It was found that glutathione reductase activity in patients with oligozoo-, astenozoo-, oligoastenozoo- and leucocytospermia were 2.3-, 1.6-, 1.7- and 3.3-fold lower than that in normozoospermic men, respectively. In addition, the glutathione transferase activity was decreased 1.8–2.1-fold in patients with oligozoo-, astenozoo- and oligoastenozoospermia and 5.0-fold in patients with leucocytospermia in comparison with healthy donors. The most significant changes were observed in the infertile men with leucocytospermia. This manifestation could be explained by the fact that white blood cells stimulate the formation of reactive oxygen species, induction and development of oxidative stress in sperm cells.
... P = 0.006 < 0.05; Figure 3). We excluded the studies of low quality (NOS ≤ 5) [53,57], yielding similar results and publication bias (SMD = 1.96, 95% CI: 1.51, 2.40, Z = 7.85, p < 0.00001; Egger's regression intercept 7.293577, t = 3.56, 95%CI 3.064078~11.52308, P = 0.002 < 0.05), so this conclusion needs to be interpreted cautiously. ...
Objectives
We conducted a systematic review and meta-analysis of observational case-control studies to evaluate markers of oxidative stress in seminal plasma of patients with male infertility.
Background
Current evidence links oxidative stress to male infertility, in which multiple markers of oxidative stress have been widely detected, publishing inconsistent results with regard to the role of oxidative stress markers in the evaluation of male infertility. Therefore, a systematic review and meta-analysis on this issue is necessary.
Results
From the 1024 articles initially retrieved, 65 studies were included in our meta-analysis with 11 oxidative stress markers, containing 3819 male infertility patients and 2012 controls. The concentrations of malondialdehyde (SMD = 1.86, p < 0.00001), NO (SMD = 0.89, p = 0.001), carbonyl protein (SMD = 2.09, p < 0.00001) in seminal plasma were significantly higher in infertility patients. The concentrations of GSH (SMD = –1.68, p < 0.00001), vitamin C (SMD = –1.12, p < 0.00001), and vitamin E (SMD = –1.48, p = 0.003), as well as the activities of catalase (SMD = –1.91, p < 0.0001), glutathione peroxidase (SMD = –1.96, p = 0.0002) and glutathione-S-transferase (SMD = –1.62, p = 0.009) declined remarkably, resulting in decreased total antioxidant capacity (SMD = –1.77, p < 0.00001). Besides, the activity of superoxide dismutase showed no statistical difference between infertility patients and controls (SMD = –0.51, p = 0.07).
Conclusions
Our meta-analysis suggests that oxidative stress in seminal plasma resulting from decreased antioxidant defense are associated with male infertility.
Methods
Using PubMed, EMBASE, CNKI, VIP, and Wanfang database, we searched for literature reporting the detection of oxidative stress markers in the seminal plasma of male infertility published up to June 2017. Standardized mean differences (SMDs) and 95% confidence intervals (95%CI) were calculated for the finally analysis.
... The lipid peroxidation cascade affects sperm membrane fluidity, which in turn impacts sperm motility. Infertile men with high malondialdehyde (MDA) levels (a by-product of lipid peroxidation) were found to have poor in vitro fertilization rates (Ben Abdallah et al. 2009). In addition, increased ROS concentrations may impair the mitochondrial membrane potential, which may then lead to the activation of the apoptotic signaling pathway in sperm cells (Wang et al. 2003). ...
Physiological levels of reactive oxygen species (ROS) are required for proper functioning of the male and female reproductive system. However, imbalances between ROS production and antioxidant systems induce oxidative stress, which can jeopardize the quality of the gamete and the developing embryo and cause many pregnancy disorders, such as spontaneous abortion, recurrent pregnancy loss, preeclampsia, fetal embryopathies, and intrauterine growth restriction. This review discusses the adverse effect of ROS-induced oxidative stress in assisted reproductive technologies (ART) outcome, ROS generated in vitro by gametes and embryos, and ROS generated by external sources in the in vitro fertilization (IVF) laboratory and the protocols used, including gamete/embryo handling, composition and pH of culture media, temperature and oxygen concentration during incubation, centrifugation and freeze-thaw protocols, as well as visible light. Studies on oral supplementation of enzymatic and nonenzymatic antioxidants are discussed. Although there is no one antioxidant that is considered the best choice for improving ART outcomes, some antioxidants show promising results. Additional well-designed trials are needed to determine the appropriate type(s) and concentration of antioxidant(s) that would be helpful to infertile patients with various etiologies. Studies are also warranted in the improvement of ART protocols to minimize ROS formation during assisted reproduction.
... DISCUSSION There is evidence to support the use of P. ginseng with its androgenic activity ( Hwang et al., 2004;Bae et al., 2012) in the treatment of male sexual dysfunction and infertility as it activates the body system that increases production of certain hormones ( Choi et al., 1995;Coon and Ernst, 2002;De Andrade et al., 2007;Jang et al., 2008 ( and its antioxidant activity. Antioxidant therapy acts as a protective mechanism against oxidative stress and improves fertility parameters ( Khaki et al., 2009) by scavenging and suppressing the formation of reactive oxygen species (ROS) and lipid peroxidation ( Ben Abdallah et al., 2009). Formaldehyde is an extremely toxic environmental contaminant that can increase the production of ROS in testicular tissues and decreased the effectiveness of the testicular antioxidant system ( Zhou et al., 2006). ...
The possible protective effects of Panax ginseng on testicular damage induced by exposure to formaldehyde were studied. Twenty-four male albino rats were divided into 4 groups (6 rats each). The first group was served as a control. The second group was exposed to 17.6 ppm of 36 % formaldehyde with the actual dynamically in an inhalation chamber for 6 hours daily for 30 days. The third group was treated with P. ginseng at a dose of 100 mg/kg body weight only for 30 days. The fourth group was treated with P. ginseng at a dose level of 100 mg/kg before formaldehyde exposure with the same exposure time and days. Testes of the experimental animals were used for biochemical, histopathological and histochemical studies. Results revealed that exposure to formaldehyde alone led to a significant increase in the testicular tissue lipid peroxidation, a decrease in reduced glutathione (GSH) content of the testes and testosterone hormones level. There were changes in blood vessels, exfoliation of spermatogenic cells in some tubules with a decrease in the length of the spermatids and vacuolation of spermatogenic layers, while using P. ginseng prior to formaldehyde exposure led to restoration of the normal structure of most of the seminiferous tubules, enhance testosterone hormones level, a decreased of the GSH levels, and increased the lipid peroxidation. In conclusion, treatment of rats with 100 mg/kg P. ginseng prior exposure to formaldehyde led to a good protection of the testes against the harmful effects induced by formaldehyde vapours.
... Various researches reported a direct correlation between the amount of lipid peroxidation in sperm and sperm dysfunction (Ben Abdallah et al., 2009). Poor penetration of the sperm into the oocyte in oligozoospermic men with high levels of ROS has been identified ( Agarwal et al., 2014). ...
Purpose:
Sperm processing (e.g., centrifugation) used in preparation for assisted reproduction can result in excessive generation of reactive oxygen species (ROS) and potential sperm damage. The use of antioxidants during sperm processing has been shown to prevent iatrogenic sperm damage, including DNA damage. In this study, we evaluated the effect of caffeic acid phenethyl ester (CAPE) on oxidative stress mediated sperm dysfunction and DNA damage.
Methods:
Semen samples were obtained to liquefy at room temperature. After centrifugation and washing protocols, spermatozoa were incubated in a single step supplemented medium with either of 10, 50 or 100 μmol/L CAPE for 2 hours at 36 °C. After incubation period, MDA levels of seminal plasma were measured. The fragmentation in sperm DNA was detected by light microscopy via use of an aniline blue assay, while ultrastructural morphology was analyzed by transmission electron microscopy.
Results:
Significant increase has been observed in percent chromatin condensation (assessed by aniline blue staining) and Malondialdehyde (Mmol/L) in oligoasthenoteratozoospermia group before the centrifugation (0.57 ± 0.15). Incubation of samples with 100 μmol/L CAPE after centrifugation resulted in a significantly lower percent chromatin condensation compared to samples incubated without CAPE (0.42 ± 0.12) (P < 0.0033). Incubation of all samples with CAPE (10 μmol/L, 50 μmol/L, 100 μmol/L.) after centrifugation resulted in a significantly lower percentage of Malondialdehyde levels.
Conclusions:
The data suggests that preincubation of spermatozoa with the antioxidant CAPE offers protection against oxidative DNA damage in vitro.
... One of used indices, able to monitor LPO is represented by malondialdehyde (MDA) which probably indicates a widespread diagnostic criterion for the oxidative damage to spermatozoa even though other markers were recently proposed [8]. MDA levels assessed both in seminal plasma and directly in spermatozoa are negatively correlated with sperm parameters [9][10][11]. Recently, our group suggested that seminal plasma MDA may represent a marker for understanding pathologies causing sperm motility decrease with particular regard to genitourinary infections and varicocele [12]. ...
There are growing evidences that the semen of infertile male population shows higher reactive oxygen species (ROS) levels concomitant with lower antioxidant capacity compared to those detected in semen of fertile population. The plasma membrane of the sperm cell, which has high levels of polyunsaturated fatty acids, renders it particularly sensitive to ROS. The aim of this study was to compare the sperm parameters (concentration, motility, morphology and vitality) and the levels of malondialdehyde (MDA), as marker of lipid peroxidation (LPO), nitric oxide (NO), ascorbic acid (AA), total (GSHt) and oxidized glutathione (GSSG) in viable sperm in a group of 38 infertile patients and in a group of 55 control subjects with unknown reproductive potential. The comparison between variables in infertile patients and controls revealed that the sperm quality was reduced in the infertile group, whereas the levels of NO, AA and GSH were significantly increased in viable spermatozoa from infertile men; however, the endogenous levels of MDA were similar in infertile and control groups. Based on our results, we could speculate that the rise of GSHt and AA levels in viable sperm of infertile group help partially to counteract the damaging effect of ROS and partly prevent a substantial LPO. The observation of the concomitant increase of NO and antioxidant indices in viable spermatozoa of infertile subjects is a novel finding and we think that these results can be useful since the viable sperm population is conceivably used in assisted reproductive technology.
... Study by Khosrowbeygi et al. [59] suggested that patient with asthenozoospermia, asthenoteratozoospermia, and oligoasthenoteratozoospermia sperm all had higher levels of PUFA and therefore, were more vulnerable to LPO caused by ROS. Another study has shown similar results, as patients with asthenozoospermia and oligoasthenoteratozoospermia also show higher MDA levels [60]. Furthermore, LPO as measured via MDA levels correlated inversely to in vitro fertilization rates in infertile men [61]. ...
Role of Oxidative Stress in Male Fertility and Idiopathic Infertility: Causes and Treatment
Epidemiological studies regarding male infertility have revealed that more and more infertile men suffer from acute or chronic inflammation of the genitourinary tract, which often occurs without any symptoms. The exact mechanism for unexplained male infertility and its correlation to oxidative stress incidence and/ or the inflammatory reactions within the male genital tract is still controversially unclear till know. Previous studies have shown the presence of cytokines such as tumor necrosis factor-α (TNF-α), a key player in the inflammation process, in the semen of infertile men. However, the mechanism of their effect on human sperm functions is still in need for further under investigations. We expect that, seminal inflammatory cytokine (TNF-α) and its expression regulator, testicular ADAM 17, will be varied, when seminal OS is established, that in turn, will vary cellular integrity and energy status. As conventional treatment of male infertility such as mineral and vitamins supplementation has poor success rates and leaves a lot to be desired, natural antioxidant ingredients are greatly suggested, nowadays, to ameliorate oxidative stress and the inflammatory detrimental effects accompanied to idiopathic male infertility. Thus, the empirical investigations of naturally occurring antioxidant modulators with prospective curative effect may support their administration as safe potent natural remedies against IMI. The role of antioxidants in the treatment of unexplained male infertility correlation to ROS and/or the inflammatory cytokines are discussed in this review.
... Various researchers reported a direct correlation between the amount of lipid peroxidation in sperm and sperm dysfunction (20,22). Poor penetration of the sperm into the oocyte in oligospermia men with high levels of ROS has been identified (23). ...
Background: Oxidative stress in reproductive system leads to sperm DNA damage and sperm membrane lipid peroxidation and may play an important role in the pathogenesis of male infertility, especially in idiopathic cases. Antioxidants such as carotenoids function against free radical damages.
Objective: The aim of this study was to determine the levels of lycopene, beta-carotene and retinol in serum and their relationship with sperm DNA damage and lipid peroxidation in infertile and normospermic males.
Materials and Methods: Sixty two infertile men and 71 normospermic men participated in this study. Blood and semen samples were collected from all subjects. Sperm DNA damage was measured using TUNEL method. Carotenoids, retinol, and malonedildehyde in serum were also determined.
Results: DNA fragmentation was higher in infertile group comparing to control group. Serum levels of lycopene, beta-carotene and, vitamin A in infertile men were significantly lower than normospermic men (p< 0.001, =0.005, and =0.003 respectively). While serum MDA was not significantly different between two groups, MDA in seminal plasma of infertile men was significantly higher than control group (p< 0.001).
Conclusion: We concluded that lycopene, beta-carotene, and retinol can reduce sperm DNA fragmentation and lipid peroxidation through their antioxidant effect. Therefore the DNA fragmentation assay and determination of antioxidants factors such as lycopene, beta-carotene and retinol, along with sperm analysis can be useful in diagnosis and treatment of men with idiopathic infertility.
... The antioxidant systems control the balance between production and neutralization of ROS and protect spermatozoa against peroxidative damage [76]. Recent studies moreover show a physiological SOD activity in human seminal plasma [77,78]. SOD is an important antioxidant defense in all cells exposed to oxygen. ...
Oxidative energy production is inevitably associated with the generation of reactive oxygen species (ROS), excessive concentrations of which can lead to cellular pathol‐ ogy. A free radical may be defined as any molecule that has one or more unpaired electrons. The superoxide anion, the hydroxyl radical, and the hypochlorite radical are some of the highest reactive radicals of oxygen. Owing to their high reactivity and to their capability of initiating an uncontrolled cascade of chain reactions, ROS produce extensive protein damage and cytoskeletal modifications and inhibit cellular mechanisms. Aerobic organisms are equipped with a powerful battery of mechanisms that protect them from the adverse effects of lipid peroxidation (LPO) and other manifestations of oxygen toxicity. Defective sperm function frequently causes male infertility, due to abnormal flagella movement, failure to recognize the zona, and inhibition of sperm-oocyte fusion. ROS are fundamental mediators of physiological sperm function, such as signal transduction mechanisms that have an effect on fertility. ROS can have positive effects on sperm and the concentration functions depending on the nature and the concentration of the ROS involved. They are necessary in regulating the hyperactivation and the ability of the spermatozoa to undergo acrosome reaction. An increased amount of superoxide anion (O 2-) is one of the first steps required by the spermatozoa for induction and development of hyperactivation and capacitation. Numerous studies have shown that oxidative stress plays an important role in the pathophysiology of infertility and assisted fertility. The paternal genome is of primary importance in the normal embryo and fetal develop‐ ment. ROS-induced sperm damage during sperm translation, such as signal trans‐ duction through the seminiferous tubules and epididymis, is one of the most important mechanisms leading to sperm DNA damage. Male germ cells are extremely vulnerable to oxidative stress as the sperm membrane is rich in unsaturated fatty acids and lacks the capacity for DNA repair. Spermatozoa are particularly susceptible to ROS-induced damage because their plasma membranes contain large quantities of polyunsaturated fatty acids (PUFA) and their cytoplasm contains low concentrations of the scavenging enzymes. Many clinical and research institutes are investigating the usefulness of antioxidant supplementation and their role in prevention of the infertility problems. Incubation under oxygen in vitro was detrimental to human spermatozoa, decreasing motility and viability. Since then, many reports have associated ROS with impaired sperm function, including decreased motility, abnormal morphology, and decreased sperm-egg penetration. Increasing knowledge of the mechanisms whereby ROS and endogenous antioxidant systems influence reproductive processes can assist to optimize the application of exogenous antioxi‐ dants to fertility treatment.
Infertility affects millions of couples worldwide and has a profound impact not only on their families, but also on communities. Telomere attrition has been associated with infertility, DNA damage and fragmentation. Oxidative stress has been shown to affect sperm DNA integrity and telomere length. Sirtuins such as SIRT1 and SIRT3 are involved in aging and oxidative stress response. The aim of the present study is to determine the role of SIRT1 and SIRT3 in regulating oxidative stress, telomere shortening, and their association with oligospermia. Therefore, we assessed the protein levels of SIRT1 and SIRT3, total antioxidant capacity (TAC), superoxide dismutase (SOD), malondialdehyde (MDA) and catalase activity (CAT) in the seminal plasma of 272 patients with oligospermia and 251 fertile men. We also measured sperm telomere length (STL) and leukocyte telomere length (LTL) using a standard real-time quantitative PCR assay. Sperm chromatin and protamine deficiency were also measured as per standard methods. Our results for oligospermic patients demonstrate significant reductions in semen parameters, shorter STL and LTL, lower levels of SOD, TAC, CAT, SIRT1 and SIRT3 levels, and also significant protamine deficiency and higher levels of MDA and DNA fragmentation. We conclude that a shorter TL in sperms and leukocytes is associated with increased oxidative stress that also accounts for high levels of DNA fragmentation in sperms. Our results support the hypothesis that various sperm parameters in the state of oligospermia are associated with or caused by reduced levels of SIRT1 and SIRT3 proteins.
This work aims to analyze Y-chromosome microdeletion in AZFc region of Oligoasthenoteratozoospermic (OAT) patients and oxidative damage profile in idiopathic OAT patients. Peripheral blood, serum, and semen samples of 57 OAT men and 57 fertile controls samples were collected for analyzing YCMD in the AZFc region using polymerase chain reaction and gel electrophoresis and to understand the oxidative damage level, total antioxidant capacity (T-AOC), Human 8-Hydroxy-deoxyguanosine (8-OHdG), Lipid peroxidation (LPO) and nitric oxide (NO) profiles were used. Out of the 57 OAT men, nine (15.7%) were observed with AZFc deletion. The remaining 48 idiopathic infertile men samples were analyzed for oxidative damage. The T-AOC levels were significantly reduced inpatient samples than in the control (p<0.05). This study shows that testing YCMD and oxidative stress damage analysis is necessary before undergoing Assisted Reproductive Technology (ART).
Background:
antioxidants that used for the infertility treatment cannot have their complete effectiveness, because of their instability in the culture medium.
Significance:
one of the most advances, in the drug delivery systems, is nanoliposomes-loaded, as biodegradable and bioavailable carriers. Hormonal and antioxidant agents encapsulating inside the nanoliposomes were used, to increase the effectiveness of antioxidants in the sperm culture medium.
Materials:
Semen sample from 15 asthenospermia were divided into 10 equal parts. After preparation, the sperms were incubated with free form of drugs and nanocarriers contained resveratrol, catalase, resveratrol-catalase and testosterone for 45 min. All sperm parameters, sperm DNA and gene expressions were evaluated before and after freezing.
Results:
Before freezing, all nanocarriers and free testosterone showed higher sperm motility compared to free drugs (P=.000). Free Testosterone and free resveratrol-catalase had higher DNA damage compared to nanocarriers (P=.000). Before freezing, the blank nanoliposome and testosterone nanoliposomes had the lowest HSP70 gene expression respectively (P = 0.005) (P = 0.001). After freezing, a significant reduction in sperm motility was observed in the free resveratrol-catalase group (P=.003). Also, a significant increase in sperm viability was observed in the free testosterone and nanoliposomes of blank and testosterone (P > 0.05). The least DNA damage was related to catalase nanoliposomes (P=.000). All nanoliposomes, especially catalase, had the highest percentage of class I morphology compared to the control group (P=.000).
Conclusions:
Nanoliposomes could improve the sperm parameters and DNA integrity before and after freezing, by increasing the effectiveness of antioxidants. So, it can be recommended in the ART lab.
Significance:
Infertility is a major global health problem, with nearly half of the cases being associated with male factors. Although reactive oxygen species (ROS) are crucial for sperm cells capacitation, an imbalance between ROS production and antioxidants can lead to oxidative stress that can impair sperm function. Indeed, high semen ROS levels are reported in 30-80% of infertile men. Recent Advances: Male oxidative stress infertility is an uprising classification for idiopathic infertility. Proteomics approaches, including quantitative mass spectrometry (MS)-based proteomics are being utilized to explore the molecular mechanisms associated with oxidative stress in male infertility.
Critical issues:
In this review, proteome data were collected from articles available on PubMed centered on MS-based proteomics studies, performed in seminal plasma and sperm cells samples, and enrolling men with impaired semen parameters. The bioinformatics analysis of proteome data with Cytoscape (ClueGO+CluePedia) and STRING tools allowed the identification of the biological processes more prevalent in asthenozoospermia, with focus on the ones related to oxidative stress.
Future directions:
The identification of the antioxidant proteins in seminal plasma and sperm cells that can protect sperm cells from oxidative stress is crucial not only for a better understanding of the molecular mechanisms associated with male infertility but especially to guide new therapeutic possibilities.
Sperm cryopreservation leads to various structural and functional damages, some of which induce by oxidative stress. The reactive oxygen species (ROS) generates by mitochondria and membrane NADPH oxidases (NOXs). Among the NOXs, only NOX5 has been identified in the cell membrane of human sperm. This study was designed to clarify the possible role of NOX5 on sperm cryoinjury. Forty human semen samples were washed and randomly divided into fresh and cryopreserved groups. Each group was divided into 4 subgroups containing Ham’s F10 (control), 0.1% DMSO (vehicle), 100 nM of PMA (phorbol 12-myristate 13-acetate) and 1 µM of DPI (diphenyleneiodonium), as NOX5 activator and inhibitor. The samples of cryopreserved groups were preserved in liquid nitrogen for 1 month. The sperm kinematics, membrane integrity, ROS production, apoptosis rate, mitochondrial membrane potential (MMP), intracellular ATP and calcium concentration [Ca2+]i were evaluated. The percent of sperm with intact membrane and motile sperm reduced significantly after thawing (p ≤ 0.01). The ROS production (p ≤ 0.01) and the apoptotic rate increased, MMP dissipated, and the percentage of live cells with high [Ca2+]i decreased significantly in the cryopreserved control group relative to the fresh control group. DPI, in contrast to PMA, improved sperm progressive motility (p ≤ 0.01), membrane integrity in fresh and cryopreserved groups and reduced the ROS amount in cryopreserved group (p ≤ 0.01). Apoptotic rate, [Ca2+]i, ATP, and MMP did not change with DPI and PMA in cryopreserved groups. We conclude that NOX5 activity in fresh sperm is low, and it increases during cryopreservation. NOX5 inhibition improves the cryopreserved sperm quality.
Significance:
Antioxidants are essential for the maintenance of cellular redox homeodynamics in the male reproductive tract, playing a key role in fertilizing potential. Reactive oxygen species (ROS), at physiological levels, are essential for sperm function and fertilization. Under pathological conditions, abnormal production of ROS may occur. Redox control is primarily regulated by the inner antioxidant system. However, these endogenous antioxidants may be present at abnormal amounts or may be insufficient. Exogenous antioxidants obtained through the diet may have an important role, particularly in specific pathological conditions. This review addresses the regulation of redox hemodynamics in the male reproductive tract by endogenous and exogenous antioxidants and the importance of their cooperation for the maintenance of fertility. Recent Advances: Many studies have shown the importance of antioxidants for the preservation of male fertility, mostly under pathological conditions. Excessive antioxidants can inhibit ROS-induced signaling pathways essential for the reproductive system. The challenge is to keep the balance between oxidants and antioxidants to manage ROS-amount at the proper physiological concentration.
Critical issues:
Although antioxidant therapies are gaining popularity and showing promising results in the improvement of male fertility, there is a lack of knowledge regarding the type of exogenous antioxidant, the doses and time to be administered.
Future directions:
It would be of great importance to find a way to restore redox homeostasis under stress conditions. Understanding the poorly studied mechanisms by which exogenous antioxidants cooperate with inner cellular antioxidant system to counteract free radicals may help in the development of new fertility therapies.
The present study was conducted to investigate
the effect of ( Sustanon
t
250;
abusing on reproductive efficiency .Thirty two male and thirty two female rats were
used'Theyweredividedintofourgroups(eachgroupconsistedofSmalesand8
females).Thefirstgroupservedascontrolgroupinwhichtheratsdosedby
intramuscular
injection of 10 p I olive oil weekly for !2 weeks '
the rest 3 treated
groups(Gl.G2andG3)inwhichtheratsdosedbyintramuscularinjectionof50.l00
and 150 mg Sustanon
t
250 / kg B.W. respectively
weekly for 12 weeks' then all rats
were terminated to determine testosterone ,FSH
and LH levels in addition to seminal
analysis . The results reveled that testosterone level was significantly
(pS 0'05)
elevated in Gr and G2 . The LH and FSH were significantly
decreased(ps
0.05) in all
animals .The seminal analysis in response to Sustanon
t
250 injection revealed a
significant
.
reduction in the sperm concentration,
viable spenn
percentage and
morphologicallynormalspermpercentage'Anothergroupof16maleand32female
rats were used ,to
determine the reproductive efficiency ,
from which 4 male and 8
femaleweredosedbyintramuscularinjectionwithl0ploliveoilweeklyfor12
rveeks,theyareconsideredascontrolanimalsTherestratsweredosedby
intramuscular
injection of.150 mg Sustanon
@
250 / kg B'W' weekly for 12 weeks '
Thentheyweredividedintofourgroups:groupl(4nontreatedmaleswithSnon
treated females ) ,
group 2 (4 treatedmales with 8 treated females )'
group 3 (4 treated
males with 8 non treated females) and group 4 (4 non treated males with 8 treated
females )
. After they were allowed to mate for 12 weeks ,
the reproductive
parameters
were investigated
which revealed
the fertility percentage as 100% in Gl 'whereasthfemale rats in G2 ,
G3 and
,
G4 showed no pregnancy at all which indicated complete
infertility.
Cryopreservation of spermatozoa is becoming more important because of new clinical requirements and current clinical practice. Despite the success of sperm cryopreservation this routinely used procedure induces serious detrimental changes in sperm function. Some researchers believe that cryopreservation is associated with DNA fragmentation and DNA single strand breaks in sperm. Mechanisms of cryodamage to human spermatozoa are thought to be multifactorial including: cold shock, osmotic stress, intracellular ice crystal formation, oxidative stress, and combinations of these conditions. Additives showing antioxidative properties reported to reduce the impact of ROS-induced and cold shock damages. Many studies exist as regards the effects of antioxidants on the cryopreservation aimed at improving the quality of post-thaw semen. Hence, this review will clarify results of recent applications of various antioxidants used in numerous research efforts to improve cryopreservation of spermatozoa. This review is to increase the understanding of the roles of these antioxidants concerning mechanisms which enhance resistance to cryodamage of spermatozoa.
Ejaculates of four crossbred boars were frozen with or without 200 μg/mL of α-tocopherol. Frozen samples were then thawed in the extender containing either ascorbic acid (2.5, 5 or 10 mM) or pyruvate (1 or 5 mM) to obtain the samples in which sperm quality parameters and lipid peroxidation level were evaluated. Ascorbic acid decreased lipid peroxidation in a dose-dependent manner and increased post-thaw sperm motility. The addition of pyruvate decreased lipid peroxidation level without improving sperm quality parameters. Although the addition of ascorbic acid or pyruvate did not show a synergic effect with α-tocopherol, the decrease of lipid peroxidation observed when the antioxidants were added to the thawing extender could be indicating a protective effect on the sperm plasma membrane from peroxidative damage. Since α-tocopherol protects from the lipid peroxidation and improves the sperm motility, it would be considered as the recommended antioxidant to use in boar sperm cryopreservation to extend the time of storage for the use in commercial applications.
Recent studies have demonstrated that human spermatozoa are capable of generating reactive oxygen species and that this activity is significantly accelerated in cases of defective sperm function. In view of the pivotal role played by lipid peroxidation in mediating free radical damage to cells, we have examined the relationships between reactive oxygen species production, lipid peroxidation, and the functional competence of human spermatozoa. Using malondialdehyde production in the presence of ferrous ion promoter as an index of lipid peroxidation, we have shown that lipid peroxidation is significantly accelerated in populations of defective spermatozoa exhibiting high levels of reactive oxygen species production or in normal cells stimulated to produce oxygen radicals by the ionophore, A23187. The functional consequences of lipid peroxidation included a dose-dependent reduction in the ability of human spermatozoa to exhibit sperm oocyte-fusion, which could be reversed by the inclusion of a chain-breaking antioxidant, alpha-tocopherol. Low levels of lipid peroxidation also had a slight enhancing effect on the generation of reactive oxygen species in response to ionophore, without influencing the steady-state activity. At higher levels of lipid peroxidation, both the basal level of reactive oxygen species production and the response to A23187 were significantly diminished. In contrast, lipid peroxidation had a highly significant, enhancing effect on the ability of human spermatozoa to bind to both homologous and heterologous zonae pellucidae via mechanisms that could again be reversed by alpha-tocopherol. These results are consistent with a causative role for lipid peroxidation in the etiology of defective sperm function and also suggest a possible physiological role for the reactive oxygen species generated by human spermatozoa in mediating sperm-zona interaction.
Cryptorchidism and oligozoospermia are clinical conditions closely associated with impaired fertility. Oxidative stress and related sperm DNA damage have been identified as significant causes of male infertility. Aim: To determine the extent of sperm nuclear DNA damage in patients affected with idiopathic oligozoospermia or undescended testes and to examine its relationship with oxidative stress. Patients and methods: We studied 20 patients with idiopathic oligozoospermia and 18 with undescended testes (who previously underwent orchiopexy)and 25 normozoospermic healthy controls. All subjects underwent semen analysis. Sperm DNA damage was evaluated by the sperm chromatin structure assay/flow cytometry (SCSA-FCM) and by the dUTP-biotin nick end labeling (TUNEL) assay. Levels of reactive oxygen species (ROS) and total antioxidant capacity (TAC) were assessed by a chemiluminescence assay. Results: DFI (percentage of sperm with denatured DNA) values and percentage of TUNEL positive cells were significantly greater in patients with oligozoospermia (DFI: 28.8±5.6; TUNEL+: 26.9±3.0) or cryptorchidism (DFI: 26.4±10.1; TUNEL+: 29.1±3.9), compared with controls (DFI: 7.1±0.9; TUNEL+: 14.2±1.2). Similarly, both groups of patients had significantly higher (p <0.01) levels of ROS. TAC levels did not differ between control and patient groups, suggesting that the DNA damage occurs before spermiation. Conclusions: Sperm DNA damage is significantly increased in men with idiopathic oligozoospermia and in cryptorchid subjects. The finding of increased ROS levels may indicate that seminal oxidative stress may be involved in the pathogenesis of sperm DNA damage in these patients Trabajo con financiamiento FONDECYT proyecto #1011045.
In view of the high content of unsaturated fatty acids in the phospholipids of mammalian spermatozoa, the possibility was investigated that during aerobic incubation of spermatozoa these fatty acids, in particular arachidonic (20:4) and docosahexanoic (22:6) acid, undergo peroxidation, which in turn is responsible for the decline in viability of spermatozoa. The formation of lipid peroxides was followed by the thiobarbituric acid (TBA) reaction, iodometry and fluorimetry; it was found that during aerobic incubation of washed ram spermatozoa at 37 degrees C in the presence of ascorbic acid (0.1 mg/ml) and ferrous iron (2.8 mu g Fe/ml), peroxides became detectable after 30 min incubation and their level reached a maximum after 60 min. Coincident with the formation of peroxides motility declined and the spermatozoa became agglutinated, but their oxygen uptake was not grossly different from that of motile spermatozoa in mixtures free of ascorbic acid or Fe. Peroxide formation could be prevented by the addition of either chelating agents, such as EDTA and sodium citrate, or antioxidants, such as tocopherol and butylated hydroxyanisole, to the incubation medium. Conclusive evidence that the substrate for the peroxidation reaction is a lipid, was obtained by extracting lipids from spermatozoa with chloroform: methanol and incubating aerobically in the presence of catalytic amounts of ascorbate and Fe, two fractions: the extracted lipid and the 'defatted' sperm residue; very high peroxide values were recorded in the former, but not the latter fraction. Examination by thin-layer and gas-liquid chromatography of the phospholipid and neutral lipid fractions obtained from spermatozoa, revealed that peroxidation caused a loss of approximately 50% in the content of plasmalogens, 20:4 and 22:6 fatty acids, and palmitaldehyde. Changes in the other phospholipids and their bound fatty acids were relatively small, but there was some increase in the amount of a substance which behaved chromatographically as lysolecithin. The content of cholesterol and glycerides in the neutral lipid fraction was not appreciably different in non-peroxidized and peroxidized spermatozoa, but the latter contained a higher proportion of free fatty acids. Our results indicate that under aerobic conditions the lipids in spermatozoa, in particular the plasmalogens, are susceptible to peroxidation. The peroxidation reaction apparently takes place within the lipoprotein, and is associated with a decline in the viability of spermatozoa. This decline could be due either to an increase in cellular permeability resulting from changes in membrane-bound lipids, or to a direct toxic effect of lipid peroxides on spermatozoa. Development of means for the prevention of adverse effects of the peroxidation reaction, may be of practical importance in relation to the problem of long-term storage of semen for artificial insemination.
Recent studies have demonstrated that human spermatozoa are capable of generating reactive oxygen species and that this activity is significantly accelerated in cases of defective sperm function. In view of the pivotal role played by lipid peroxidation in mediating free radical damage to cells, we have examined the relationships between reactive oxygen species production, lipid peroxidation, and the functional competence of human spermatozoa. Using malondialdehyde production in the presence of ferrous ion promoter as an index of lipid peroxidation, we have shown that lipid peroxidation is significantly accelerated in populations of defective spermatozoa exhibiting high levels of reactive oxygen species production or in normal cells stimulated to produce oxygen radicals by the ionophore, A23187. The functional consequences of lipid peroxidation included a dose-dependent reduction in the ability of human spermatozoa to exhibit sperm oocyte-fusion, which could be reversed by the inclusion of a chain-breaking antioxidant, alpha-tocopherol. Low levels of lipid peroxidation also had a slight enhancing effect on the generation of reactive oxygen species in response to ionophore, without influencing the steady-state activity. At higher levels of lipid peroxidation, both the basal level of reactive oxygen species production and the response to A23187 were significantly diminished. In contrast, lipid peroxidation had a highly significant, enhancing effect on the ability of human spermatozoa to bind to both homologous and heterologous zonae pellucidae via mechanisms that could again be reversed by alpha-tocopherol. These results are consistent with a causative role for lipid peroxidation in the etiology of defective sperm function and also suggest a possible physiological role for the reactive oxygen species generated by human spermatozoa in mediating sperm-zona interaction.
Reactive oxygen species (ROS) have beneficial or detrimental effects on sperm functions depending on the nature and the concentration
of the ROS involved, as well as the moment and the location of exposure. Excessive generation of ROS in semen, mainly by neutrophils
but also by abnormal spermatozoa, could be a cause for infertility. Hydrogen peroxide is the primary toxic ROS for human spermatozoa.
Low concentrations of this ROS do not affect sperm viability but cause sperm immobilization mostly via depletion of intracellular
ATP and the subsequent decrease in the phosphorylation of axonemal proteins. High concentrations of hydrogen peroxide induce
lipid peroxidation and result in cell death. On the other hand, the superoxide anion appears to play a major role in the development
of hyperactivation and capacitation. The observations that: (i) exogenously generated superoxide anions induce hyperactivation
and capacitation; (ii) capacitating spermatozoa themselves produce elevated concentrations of superoxide anion over prolonged
periods of time; and (iii) removal of this ROS by superoxide dismutase prevents sperm hyperactivation and capacitation induced
by various biological fluids, stress the importance of the superoxide anion in these processes.
We attempted to determine reactive oxygen species production by the spermatozoa of patients with idiopathic infertility and healthy donors, and observe whether increased production was due to decreased seminal plasma reactive oxygen species scavengers.
Reactive oxygen species production by spermatozoa and seminal plasma antioxidants was assayed in 18 patients with idiopathic infertility and 10 controls. Reactive oxygen species formation and seminal plasma antioxidants were measured by luminol and lucigenin dependent chemoluminescence, and enzymatic methods, respectively.
Higher reactive oxygen species production was observed in 16 of the 18 patients (88.8%, p < 0.0001 versus controls). Seminal plasma superoxide dismutase, catalase, glutathione peroxidase and total sulfhydryl group levels in infertile patients were significantly lower than in controls.
Decreased seminal plasma antioxidant activity and increased reactive oxygen species production can be responsible for idiopathic male infertility.
Mammalian spermatozoa are rich in polyunsaturated fatty acids and are very susceptible to attack by reactive oxygen species (ROS) and membrane lipid peroxide ion. Normally a balance is maintained between the amount of ROS produced and that scavenged. Cellular damage arises when this equilibrium is disturbed. A shift in the levels of ROS towards pro-oxidants in semen and vaginal secretions can induce an oxidative stress on spermatozoa. The aim was to study lipid peroxidation and antioxidant enzymes such as catalase, glutathione peroxidase and superoxide dismutase (SOD) and to correlate the same, with the 'water test', in male infertility.
Experimental study.
Ejaculates from a total of 83 infertile and fertile healthy individuals were obtained. Lipid peroxidation and antioxidant enzyme levels were studied and correlated with water test.
The results indicate that (i) the antioxidant enzyme catalase showed no significant changes in the various pathological samples, (ii) antioxidant enzymes SOD and glutathione peroxidase correlate positively with asthenozoospermic samples and (iii) the degree of lipid peroxidation also correlates positively with the poorly swollen sperm tails. The increase in SOD and glutathione peroxidase values, in the pathological cases represents an attempt made to overcome the reactive oxygen species.
Water test could be used as a preliminary marker test for sperm tail damage by reactive oxygen species, since it correlates very well with lipid peroxidation and antioxidant enzymes.
There is growing evidence that damage to spermatozoa by reactive oxygen species (ROS) play a key role in male infertility. The aim of the present study was to assess seminal plasma levels of total antioxidant capacity (TAC), free 8-Isoprostane and activities of catalase and superoxide dismutase (SOD) in men with asthenozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia compared with normozoospermic males.
The patients consisted of 46 men with seminal parameters abnormalities. The patients were grouped into asthenozoospermic (n = 15), asthenoteratozoospermic (n = 16) and oligoasthenoteratozoospermic (n = 15). The control group consisted of 16 healthy males with normozoospermia. Catalase activity was measured by Aebi spectrophotometeric method. Levels of TAC and SOD were measured by commercially available colorimetric assays. Level of free 8-Isoprostane was assessed by commercially available enzyme immunoassay (EIA) method. Differences between groups were assessed using Mann-Whitney U test and Kruskal-Wallis test. Coefficients of correlation were calculated using Spearman's correlation analysis. All hypothesis tests were two-tailed with statistical significance assessed at the p value < 0.05 level with 95% confidence intervals.
Levels of catalase and TAC were significantly lower in patients than the control group. No significant changes were seen in SOD activities. Levels of free 8-Isoprostane were significantly higher in patients than the control group. Furthermore, asthenozoospermic, asthenoteratozoospermic and oligoasthenoteratozoospermic groups had significantly lower values of catalase activity and TAC when compared to normozoospermic males. Levels of free 8-Isoprostane were significantly higher in all patients subgroups than the control group. Levels of catalase and TAC were positively correlated with sperm motility and morphology. Free 8-Isoprostane levels showed an inverse correlation with sperm motility and morphology.
Decreasing seminal plasma antioxidants levels, especially catalase and TAC, could have significant role in etiology of impaired sperm function. Measurement of 8-Isoprostane may be used as a specific biomarker for assessing oxidative stress on sperm.
The autoxidation of pyrogallol was investigated in the presence of EDTA in the pH range 7.9–10.6.
The rate of autoxidation increases with increasing pH. At pH 7.9 the reaction is inhibited to 99% by superoxide dismutase, indicating an almost total dependence on the participation of the superoxide anion radical, O2·−, in the reaction. Up to pH 9.1 the reaction is still inhibited to over 90% by superoxide dismutase, but at higher alkalinity, O2·− -independent mechanisms rapidly become dominant.
Catalase has no effect on the autoxidation but decreases the oxygen consumption by half, showing that H2O2 is the stable product of oxygen and that H2O2 is not involved in the autoxidation mechanism.
A simple and rapid method for the assay of superoxide dismutase is described, based on the ability of the enzyme to inhibit the autoxidation of pyrogallol.
A plausible explanation is given for the non-competitive part of the inhibition of catechol O-methyltransferase brought about by pyrogallol.
Background: Reactive Oxygen Species (ROS) are a group of free radicals that in excessive amounts have negative influence on sperm quality and function. Objective: We compared ROS levels in seminal plasma of infertile men with this level in healthy donors. We also determined the ROS level in semen of infertile men according to the etiology of infertility, and also the effect of smoking on its level.Materials and Methods: In total, 63 infertile patients and 63 healthy donors as control were selected. Complete physical examination, semen analysis, scrotal sonography and hormone assay were done for all patients. Azoospermic patients were excluded from the study. ROS level in semen was measured by chemiluminescence assay in both groups. Patients also were divided in two groups according to the etiology of infertility.Results: The mean age of normal subjects and infertile men were 30.78±3.73 years and 31.43±6.60 years respectively. The mean ROS level in normal men was 180.05 RLU (Relative light unit), while this was 1852.04 RLU in infertile patients, which is significantly higher in case group (p=0.000). Fifty patients had varicocele and in 13 patients no specific etiology was found. The mean ROS level in varicocele patients was 2215.42 RLU and in unknown group was 454.44 RLU (p=0.048).Conclusion: Our study showed that the level of ROS in seminal fluid of infertile men is significantly higher than fertile donors and also in infertile patients with varicocele it is higher than patients with unknown cause.
To investigate the effects of 17beta-estradiol (E2), Peganum harmala extract (PHE) and caloric restriction (CR) on various testis parameters during aging.
Twelve month-old male rats were treated for 6 months with either E2 or PHE, or submitted to CR (40%).
Our results show that estrogens and CR are able to protect the male gonad by preventing the decrease of testosterone and E2 levels as well as the decrease of aromatase and estrogen receptor gene expressions. Indeed, E2, PHE and CR treatments induced an increase in the superoxide dismutase activities and decreased the activity of testicular enzymes: gamma-glutamyl transferase, alkaline phosphatase, lactate deshydrogenase as well as the aspartate and lactate transaminases in aged animals. In addition, the testicular catalase and gluthatione peroxidase activities were enhanced in E2, PHE and CR-treated rats compared to untreated animals at 18 months of age. Moreover, the positive effects of estradiol, PHE and CR were further supported by a lower level of lipid peroxidation. Recovery of spermatogenesis was recorded in treated rats.
Besides a low caloric diet which is beneficial for spermatogenesis, a protective antioxydant role of estrogens is suggested. Estrogens delay testicular cell damage, which leads to functional senescence and, therefore, estrogens are helpful in protecting the reproductive functions from the adverse effects exerted by reactive oxygen species (ROS) produced in large quantities in the aged testis.
Aerobic incubation of human spermatozoa in the presence of catalytic amounts of ascorbate and ferrous ion results in rapid peroxidative breakdown of sperm phospholipids and fatty acids; most strongly affected are phosphatidyl ethanolamine, ethanolamine plasmalogen, and docosahexanoic acid. Both peroxidation of the endogenous sperm phospholipid and the concurrent loss of motility can be fully prevented, but not reversed, by an "antiperoxidant" factor present in human seminal plasma. Exogenously applied lipid peroxides are powerfully spermicidal. Washed human spermatozoa, at a concentration normally present in semen, treated with as little as 30 nmoles of lipid peroxide/ml become irreversibly immotile within a few minutes. The antiperoxidant factor present in human seminal plasma effectively counteracts the toxic effect of exogenous peroxidized fatty acids upon human spermatozoa, but is unable to restore motility lost by lipid peroxide action.
To investigate whether activated poly-morphonuclear neutrophils (PMN) can affect sperm motility and to assess the capacity of seminal plasma and of scavengers of reactive oxygen species to prevent any loss of motility.
Blood PMN were isolated, and their capacity to generate reactive oxygen species was investigated. The effect of activated PMN was tested on the motility of Percoll-washed spermatozoa in the presence and absence of reactive oxygen species scavengers or seminal plasma (whole or fractionated).
Fertile volunteers and patients attending the Infertility Clinic at the Royal Victoria Hospital.
The production of reactive oxygen species by activated PMN occurred in the 1st hour after stimulation and was linearly correlated with the concentration of PMN. Percoll-washed spermatozoa had reduced motility in the presence of PMN, and the drop in motility was more severe as the concentration of PMN was increased. Catalase and dimethylsulfoxide reduced the toxic effect of PMN on sperm motility, whereas superoxide dismutase was without effect. Seminal plasma (50%, vol/vol) allowed a highly variable protective effect against the loss of sperm motility that appeared to originate predominantly from the low molecular weight (< 10 kd) fraction but also from the high molecular weight (> 12 kd) fraction of seminal plasma.
These data suggest that [1] the presence of PMN, even at 0.6 x 10(6) cells/mL, in semen can be hazardous for spermatozoa, [2] H2O2 and .OH are responsible for the loss of motility, and [3H] seminal plasma confers a variable protection due to high and low molecular weight factors.
Catalase activity was determined in human semen by measuring the oxygen burst with a Clark electrode, after H2O2 addition. Significant catalase activities (mean +/- SD) were found in migrated, motile spermatozoa (44 +/- 17 nmoles O2/min/10(8) cells) and in seminal plasma of normozoospermic men (129 +/- 59 nmoles O2/min/ml). It has been demonstrated that seminal catalase originated from prostate; however, its activity was not correlated with the usual prostatic markers (such as citric acid and zinc). Our data suggest a multiglandular function secreted by this organ. The catalase activities measured in seminal samples from asthenozoospermic, infertile men were found lower than those from normozoospermic subjects. The understanding of the relative contribution of the different enzyme systems against O2 toxicity (superoxide dismutase, catalase, glutathione peroxidase) seem to be a priority area of research to understand disturbances of sperm function.
Because the amounts of lipid peroxides in the blood are rather small, a sensitive assay method is needed. For this purpose, the most appropriate among several reactions for detecting lipid peroxides is the thiobarbituric acid (TBA) reaction because of its sensitivity. TBA reaction with lipid peroxides gives a red-colored pigment. Malondialdehyde also gives the same product upon reaction with TBA. Because this product is fluorescent, a sensitive assay can be made by fluorometry. Results show that lipid peroxides can be measured by TBA reaction with fluorometry. Elimination of TBA-reacting substances other than lipid peroxides is necessary for the measurement of lipid peroxides in serum. The elimination procedure must be simple to avoid artifact due to the peroxidation during the procedure. One of the best procedures is to isolate lipids by precipitating them along with serum protein with the phosphotungstic acid–sulfuric acid system. By this procedure, water-soluble substances, which react with TBA to yield the same product as lipid peroxides, are removed. It was anticipated that platelet aggregation, if it occurs during the drawing of the blood, would liberate the TBA-reacting substances, and the effect of the aggregation was found to be eliminated by treatment with phosphotungstic acid–sulfuric acid system.
Publisher Summary Catalase exerts a dual function: (1) decomposition of H 2 O 2 to give H 2 O and O 2 (catalytic activity) and (2) oxidation of H donors, for example, methanol, ethanol, formic acid, phenols, with the consumption of 1 mol of peroxide (peroxide activity). The kinetics of catalase does not obey the normal pattern. Measurements of enzyme activity at substrate saturation or determination of the K s is therefore impossible. In contrast to reactions proceeding at substrate saturation, the enzymic decomposition of H 2 O 2 is a first-order reaction, the rate of which is always proportional to the peroxide concentration present. Consequently, to avoid a rapid decrease in the initial rate of the reaction, the assay must be carried out with relatively low concentrations of H 2 O 2 (about 0.01 M). This chapter discusses the catalytic activity of catalase. The method of choice for biological material, however, is ultraviolet (UV) spectrophotometry. Titrimetric methods are suitable for comparative studies. For large series of measurements, there are either simple screening tests, which give a quick indication of the approximative catalase activity, or automated methods.
The mechanisms by which ferrous ion promoters induce malondialdehyde generation by human spermatozoa have been investigated in order to provide a rational basis for the quantification and interpretation of lipid peroxidation assays. Incubation of human spermatozoa with a ferrous ion promoter in the presence of thiobarbituric acid (TBA) led to the generation of the bone fide malondialdehyde-TBA adduct. The importance of iron in the stimulation of lipid peroxidation was emphasized by the ability of Desferal and EDTA to suppress malondialdehyde generation. Paradoxically, when the concentration of EDTA relative to iron was equimolar or greater, the suppression of malondialdehyde formation was accompanied by the generation of hydroxyl radicals. These results suggested that the addition of promoter did not effect the first-chain initiation of lipid peroxidation but favored an alternative mechanism involving the catalytic decomposition of pre-existing lipid peroxides. This conclusion was reinforced by the inability of reagents that would limit the formation (superoxide dismutase and/or catalase) or availability (mannitol, formate) of hydroxyl radicals, to influence malondialdehyde generation. While hydroxyl radicals were not directly involved in Fe(2+)-promoted malondialdehyde generation, the existence of significant correlations between reactive oxygen species production and the outcome of the TBA assay, suggested that Fenton chemistry might be important in the initiation of peroxidative damage. It is proposed that the impeded propagation of peroxidation initiated by Fenton or Haber Weiss reactions would lead to the accumulation of lipid peroxides in the spermatozoa and it is these peroxides that are induced to decompose during the Fe(2+)-promoted TBA assay, stimulating a lipoperoxidative chain reaction and malondialdehyde formation.
Intact human sperm incorporated radiolabelled fatty acids into membrane phospholipids when incubated in medium containing bovine serum albumin as a fatty acid carrier. The polyunsaturated fatty acids were preferentially incorporated into the plasmalogen fraction of phospholipid. Uptake was linear with time over 2 hr; at this time sufficient label was available to determine the loss of fatty acids under conditions of spontaneous lipid peroxidation. Loss of the various phospholipid types, the loss of the various fatty acids from these phospholipids, and the overall loss of fatty acids were all first order. The loss of saturated fatty acids was slow with first order rate constant k1 = 0.003 hr-1; for the polyunsaturated fatty acids, arachidonic and docosahexaenoic acids, k1 = 0.145 and 0.162 hr-1, respectively. The rate of loss of fatty acids from the various phospholipid types was dependent on the type, with loss from phosphatidylethanolamine being the most rapid. Among the phospholipid types, phosphatidylethanolamine was lost at the greatest rate. Analysis of fatty acid loss through oxidation products was determined for radiolabelled arachidonic acid. Under conditions of spontaneous lipid peroxidation at 37 degrees C under air in the absence of albumin, free arachidonic acid was found in the medium, along with minor amounts of hydroxylated derivative. All the hydroperoxy fatty acid remained in the cells. In the presence of albumin, all the hydroperoxy fatty acid was found in the supernatant bound to albumin; none could be detected in the cells. Albumin is known as a very potent inhibitor of lipid peroxidation in sperm; its action may be explained, based on these results, as binding the damaging hydroperoxy fatty acids. These results also indicate that a phospholipase A2 may act in peroxidative defense by excising a hydroperoxy acyl group from phospholipid and providing the hydroperoxy fatty acid product as substrate to glutathione peroxidase. This formulation targets hydroperoxy fatty acid as a key intermediate in peroxidative degradation.
Asthenospermia is the main factor of male infertility among patients consulting the Asir Infertility Center in Abha, Saudi Arabia. Lipid peroxidation occurring in both the seminal plasma and spermatozoa was estimated by malondialdehyde (MDA) concentration. Spermatozoal MDA concentration was higher in men with decreased sperm motility. The MDA concentration in the seminal plasma exhibited no relationship with sperm concentration, sperm motility, the number of immotile spermatozoa, or even the absence of spermatozoa. The MDA concentration in sperm pellet suspensions of asthenospermic and oligoasthenospermic patients was almost twice that of the normospermic males. The MDA concentration in the sperm pellet suspension from normospermic or oligospermic patients was about 10% that in the seminal plasma. However, the MDA concentration in the sperm pellet suspension of asthenospermic or oligoasthenospermic patients was about 15% that in the seminal plasma. Treatment of asthenospermic patients with oral Vitamin E significantly decreased the MDA concentration in spermatozoa and improved sperm motility. Eleven out of the 52 treated patients (21%) impregnated their spouses; nine of the spouses successfully ended with normal term deliveries, whereas the other two aborted in the first trimester. No pregnancies were reported in the spouses of the placebo-treated patients.
Human spermatozoa possess a specialized capacity to generate reactive oxygen species (ROS) that is thought to be of significance in the redox regulation of sperm capacitation (De Lamirande and Gagnon, 1993; Aitken et al., 1995). However, the mechanisms by which ROS are generated by these cells are not understood. In this study we have examined the possible significance of NADPH as a substrate for ROS production by human spermatozoa. Addition of NADPH to viable populations of motile spermatozoa induced a sudden dose-dependent increase in the rate of superoxide generation via mechanisms that could not be disrupted by inhibitors of the mitochondrial electron transport chain (antimycin A, rotenone, carbonyl cyanide m-chlorophenylhydrazone [CCCP], and sodium azide), diaphorase (dicoumarol) xanthine oxidase (allopurinol), or lactic acid dehydrogenase (sodium oxamate). However, NADPH-induced ROS generation could be stimulated by permeabilization and was negatively correlated with sperm function. Both NADH and NADPH were active electron donors in this system, while NAD+ and NADP+ exhibited little activity. Stereo-specificity was evident in the response in that only the beta-isomer of NADPH supported superoxide production. The involvement of a flavoprotein in the electron transfer process was indicated by the high sensitivity of the oxidase to inhibition by diphenylene iodonium and quinacrine. These results indicate that NAD(P)H can serve as an electron donor for superoxide generation by human spermatozoa and present a simple strategy for the production of motile populations of free radical generating cells with which to study the significance of these molecules in the control of normal and pathological sperm function.
Reactive oxygen species, which are primarily produced by leukocytes, are generally detrimental to sperm. High reactive oxygen species levels are found in men with abnormal sperm function. Since men often have poor sperm characteristics and infertility after vasectomy reversal, we compared reactive oxygen species in seminal cells of men after vasovasostomy to those of fertile men to determine if reactive oxygen species were elevated in the former group.
We studied semen samples of men with proved fertility (39) and those with previously proved fertility who had undergone vasectomy reversal (45). The presence of leukocytes was determined by Bryan-Leishman staining. Reactive oxygen species endogenous activity was monitored by luminol dependent chemiluminescence in washed cells, including all cells in the semen, and Percoll density gradient purified sperm.
After vasovasostomy men had significantly lower sperm concentration, motility and computerized motility measurements than fertile men. Mean reactive oxygen species in washed seminal cells after vasovasostomy was 684 relative light units per second compared to 49 for fertile controls (p < 0.0001). Density gradient purified sperm had 53 and 0.64 relative light units per second, respectively (p < 0.0001). When men with leukocytospermia were excluded from analysis, differences between the groups remained, although 9 times more reactive oxygen species were detected in men after vasectomy reversal with than those without leukocytes in semen.
Higher levels of reactive oxygen species are found in washed seminal cells and purified sperm after vasectomy reversal than in those of fertile men. Although leukocytes are probably a significant source of reactive oxygen species in these groups, they may not account for all of the increased reactive oxygen species after vasovasostomy. Low motility after vasectomy reversal may be related to the detrimental effects of reactive oxygen species produced by leukocytes or sperm, even in men without clinical leukocytospermia.
To evaluate seminal oxidative stress in men after vasectomy reversal and to determine whether seminal oxidative stress could predict fertility after vasectomy reversal.
Measurement of seminal reactive oxygen species (ROS) and total antioxidant capacity (TAC) in normal donors, men who were fertile after vasectomy reversal, and men who were infertile after vasectomy reversal.
A male infertility clinic of a tertiary care center.
Thirty men who underwent vasectomy reversal and 17 normal donors.
None.
Semen characteristics, seminal ROS, and TAC were measured with chemiluminescence assays in samples from donors and reversal patients.
Mean adjusted seminal ROS (log [ROS+1]) was higher in infertile reversal patients (2.38+/-0.25) than in normal donors (1.30+/-0.14). Seminal ROS was also higher in all (fertile and infertile reversal combined) reversal patients than in donors. Total antioxidant capacity did not differ between groups. The ROS-TAC score, a composite index of seminal oxidative stress, was a significant predictor of fertility. A ROS-TAC score of 45 or greater had a positive predictive value of 73% in predicting fertility.
Seminal oxidative stress is associated with vasectomy reversal. The ROS-TAC score is a possible predictor of infertility after vasectomy reversal.
To evaluate catalase- and superoxide dismutase (SOD)-like activities in the seminal plasma of fertile and infertile men.
Semen samples were obtained from consecutive men presenting for vasectomy (n = 12) and infertility evaluation (n = 105) at our institution. Catalase-like activity was measured by the decrease in hydrogen peroxide after incubation with seminal plasma. SOD-like activity was measured as the inhibition of nitroblue tetrazolium reduction due to superoxide anion generation by xanthine plus xanthine oxidase.
Mean seminal catalase-like activity (+/-SEM) in fertile men was not significantly different from that of infertile men (369 +/- 49 versus 326 +/- 17 U/mL, respectively). Mean SOD-like activity in the semen of infertile men was significantly greater than in the semen of fertile controls (46.7 +/- 1.5 versus 37.0 +/- 2.8 U/mL, respectively, P <0.05).
Our data show that infertile men do not have deficient seminal plasma SOD- and/or catalase-like activity (two key antioxidants). These findings suggest that the high semen ROS levels in some infertile men are likely due to excessive generation of ROS rather than deficient ROS scavenging activity in semen.
In the present study, the concentrations of copper, iron, zinc, and malondialdehyde in human seminal plasma were measured and correlated with the sperm count and motility in human semen. Copper, iron, and zinc were analyzed by atomic absorption spectrometry, whereas malondialdehyde was measured by high-performance liquid chromatography. The malondialdehyde concentrations in asthenospermia and oligoasthenospermia were significantly higher than in normospermia. Copper and iron levels were higher in asthenospermia, whereas the zinc concentrations in both oligospermia and asthenospermia were lower than in normal controls. A negative correlation (r = -0.28, p < 0.05) between the malondialdehyde concentration and sperm motility was observed in the abnormal groups. There was no association among copper, iron, zinc, and malondialdehyde in seminal plasma. We concluded that changes in trace elements may be related to sperm quality and that lipid peroxidation, although it is not promoted in the seminal plasma by copper or iron or ameliorated by zinc, may be involved in the loss of sperm motility.
We have studied the lipid peroxidation product (malondiadehyde-MDA) levels after thiobarbituric acid reaction, spectrophotometrically and by a high-performance liquid chromatography (HPLC) with UV detection in seminal plasma as well as in the cell fraction. Semen samples were obtained from healthy volunteers and infertile males. Ejaculates were previously classified into studied subgroups according to standard andrological criteria (sperm number, motility, morphology) and defined as: normozoospermia (K), azoospermia (Az), teratozoospermia (T), asthenoteratozoospermia (AT), oligoasthenoteratozoospermia (OAT) and idiopathic infertility (NIF). MDA is a good marker for lipid peroxidation. There were found elevated MDA concentrations (determined by HPLC) in seminal plasma of the all analysed pathological semen samples, especially in patients with OAT. The more pronounced differences between healthy controls and NIF patients were observed in intracellular compartment. The lipid peroxidation of rich in unsaturated fatty acids sperm membranes is considered to be one of the most important effects from ROS-induced cell damage what may lead to persistent infertility.
Abnormal reactive oxygen species (ROS) production is associated with defective sperm function. Superoxide dismutase (SOD) is related with the scavenging of seminal ROS. We aimed to determine the effect of SOD activities of spermatozoa and seminal plasma on sperm quality. Semen samples from infertile couples who consented to the analyses were divided into two groups: 1) normospermia (n = 20); and 2) oligoasthenozoospermia (n = 31). The SOD activities of the spermatozoa and seminal plasma were measured by determining the inhibition of pyrogallol autoxidation. The SOD activities of spermatozoa and seminal plasma in both groups were compared. The relationships between the SOD activities and the sperm qualities were determined. We noted that SOD activities of sperm/seminal plasma in both groups were nonsignificantly different (group 1 vs. 2 = 0.77 +/- 0.33/0.84 +/- 0.40 U/mg protein for sperm, and 0.66 +/- and 0.36/0.83 +/- 0.47 U/ml for seminal plasma). SOD activities of sperm/seminal plasma were positively but nonsignificantly correlated with the sperm motility (SOD of sperm = 0.0008 x motility + 0.67; SOD of seminal plasma = 0.0006 x motility + 0.81) and concentration (SOD of sperm = 0.0006 x concentration + 0.67; SOD of seminal plasma = 0.0021 x concentration + 0.73). We concluded that SOD activities of sperm and seminal plasma were nonsignificantly correlated with the seminal quality. It appears that the SOD survey is not a useful tool for determining sperm fertilization potential.
Numerous studies have reported beneficial effects of antioxidant drugs on semen quality, but there is no well-defined therapeutical protocol in male infertility. This study aimed to test the effects of vitamin E and selenium supplementation on lipid peroxidation and on sperm parameters. The study included 54 voluntary and infertile men who produced semen samples for spermiogram and for spectrophotometric measurement of a lipid peroxidation marker, the malondialdehyde (MDA), and produced blood samples for high-performance liquid chromatography assessment of serum vitamin E level. The trial was randomized and open. Twenty-eight men were supplemented daily by vitamin E (400 mg) and selenium (225 microg), during 3 months. The remaining 26 patients received vitamin B (4,5 g/day) for the same duration. Only 20 patients achieved their treatment and returned for control analysis. MDA concentrations in sperm were much less than in seminal plasma and motility and viability were inversely correlated with semen MDA levels. In contrast to vitamin B supplementation, vitamin E and selenium supplementation produced a significant decrease in MDA concentrations and an improvement of sperm motility. The results confirm the protective and beneficial effects of vitamin E and selenium on semen quality and advocate their use in male infertility treatment.
To investigate the level of malondialdehyde (MDA), a direct indicator of lipid peroxidation-induced injury by reactive oxygen species (ROS), in testicular biopsy specimens from infertile patients.
Levels of MDA were measured in testicular biopsy specimens from 29 consequent-randomized infertile men, aged 29.58+/-4.76 (21 to approximately 45) years. All patients were evaluated by a complete medical and reproductive history, physical examination, semen analysis (at least two), serum follicle-stimulating hormone and free testosterone levels, testicular biopsy and contact imprint. Scrotal colour Doppler ultrasonography was used to confirm suspected varicocele. The testicular MDA level was measured using the thiobarbituric acid test and the results were expressed per unit tissue weight.
As a causal factor in infertility, varicocele was identified in 17 (58.6 %) patients, and idiopathic infertility, testicular failure and obstruction in 4 (13.8 %) patients each. The testicular MDA level was 13.56 (6.01), 49.56 (24.04), 58.53 (48.07), and 32.64 (21.51), 32.72 (13.61), 23.07 (7.82), 42,12 (34.76) pmol/mg tissue in the normal spermatogenesis (control), late maturation arrest, Sertoli cell only (SCO) and hypospermatogenesis (mild, moderate, severe) groups, respectively. The elevation of MDA levels was significant in the testicular tissue from SCO and maturation arrest groups compared with the controls (P<0.05). In addition, the elevation in testicular MDA levels between the SCO and the moderate hypospermatogenesis, and the moderate hypospermatogenesis and the maturation arrest groups was significant (P<0.05).
Severe pathologic changes in the testicular tissue are associated with a high level of lipid peroxidation. These findings suggest that overproduction of ROS may play a role in the mechanism of testicular degeneration associated with infertility.
The generation of reactive oxygen species (ROS) has been implicated in the regulation of sperm capacitation and acrosome reaction; however, the mechanisms underlying this regulation remain unclear. To examine the cellular processes involved, we studied the effect of different concentrations of hydrogen peroxide (H(2)O(2)) on protein tyrosine phosphorylation under various conditions. Treatment of spermatozoa with H(2)O(2) in medium without heparin caused a time- and dose-dependent increase in protein tyrosine phosphorylation of at least six proteins in which maximal effect was seen after 2 h of incubation with 50 microM H(2)O(2). At much higher concentrations of H(2)O(2) (0.5 mM), there is significant reduction in the phosphorylation level, and no protein tyrosine phosphorylation is observed at 5 mM H(2)O(2) after 4 h of incubation. Exogenous NADPH enhanced protein tyrosine phosphorylation similarly to H(2)O(2). These two agents, but not heparin, induced Ca(2+)-dependent tyrosine phosphorylation of an 80-kDa protein. Treatment with H(2)O(2) (50 microM) caused approximately a twofold increase in cAMP, which is comparable to the effect of bicarbonate, a known activator of soluble adenylyl cyclase in sperm. This report suggests that relatively low concentrations of H(2)O(2) are beneficial for sperm capacitation, but that too high a concentration inhibits this process. We also conclude that H(2)O(2) activates adenylyl cyclase to produce cAMP, leading to protein kinase A-dependent protein tyrosine phosphorylation.
Human ejaculated sperm comprised discrete subsets of spermatozoa, with different degrees of maturation. These subpopulations can be isolated through density gradient centrifugation. Sperm from the lowest density layer show the highest content of docosahexaenoic acid and sterols, and produce the highest levels of reactive oxygen species. The main objective of this study was to determine the superoxide dismutase (SOD) content and fatty acid composition of subsets of spermatozoa isolated from normozoospermic, asthenozoospermic, and polyzoospermic semen samples. Four sperm fractions (1-4) were obtained using ISolate gradient centrifugation. Morphology, motion parameters, SOD content, and fatty acid composition were assessed in the original samples and their fractions. Overall, sperm from normozoospermic samples had higher SOD content than those of asthenozoospermic or polyzoospermic samples. Once fractionated in subsets, the sperm SOD content decreased significantly (P < 0.0001) from fraction 1 (top) to 4 (bottom) in all three groups of samples. Fatty acid content as well as the oxidation coefficient followed the same pattern, decreasing from fraction 1 to 4 (F1-F4). Normo- and polyzoospermic samples showed similar amounts of fatty acids, while asthenozoospermic samples mostly revealed increased levels. Normozoospermic samples displayed the lowest unsaturated fatty acid (UFA)/SOD ratio. Spermatozoa from astheno- and polyzoospermic samples, two common seminal pathologies, showed higher UFA and lower SOD content than normal sperm, therefore exhibiting a higher susceptibility to peroxidative damage. F4 from all groups, containing the most mature spermatozoa, displayed the lowest polyunsaturated fatty acid and SOD content of all subsets, suggesting that excessive SOD activity as well as abundant peroxidative targets may both be deleterious to sperm function.
To examine the relationship between sperm reactive oxygen species (ROS) production and sperm morphology in a group of infertile men and healthy fertile donors.
A prospective clinical study.
Male infertility clinic, Glickman Urological Institute, The Cleveland Clinic Foundation, Cleveland, Ohio, and the Reproductive Medicine Unit, Liverpool Women's Hospital, United Kingdom
Thirty-nine infertile men and 13 healthy fertile donors (control).
Standard semen analysis, seminal leukocyte concentration, assessment of sperm morphology, and measurement of sperm ROS production.
Levels of sperm ROS production, percentages of different sperm morphological abnormalities, and the sperm deformity index (SDI) scores.
A significant negative correlation was observed between sperm ROS production and the proportion of sperm with normal morphology and borderline morphology. Reactive oxygen species production was positively correlated with the proportion of sperm with amorphous heads, damaged acrosomes, midpiece defects, cytoplasmic droplets, tail defects, and SDI scores. Logistic regression analysis identified a two-variable model including SDI and percentage sperm motility, which correctly identified 84% of individuals with high seminal ROS and 85% of individuals with low seminal ROS. The model had an overall accuracy of 85%.
The standard semen analysis to assess sperm motility, sperm morphology, and the SDI scores is a useful tool in identifying infertile men with high seminal ROS in infertility clinics where facilities for measuring levels of seminal ROS are not available.
A range of compounds with a role in oxidative stress were measured in ejaculates from 40 normozoospermic individuals and 93 infertile males. Ejaculates were classified according to WHO criteria. Seminal plasma and the sperm cell fraction were assessed separately for superoxide dismutase (SOD), catalase, xanthine oxidase, capability for singlet oxygen trapping and content of thiobarbituric acid-reactive substances (TBARS). Pathological cases defined as oligozoospermia, asthenozoospermia, or teratozoospermia revealed different backgrounds of oxidative stress as reflected by different levels of tested substances in every type of sperm pathology. In the majority of abnormal ejaculates, a significant increase in intracellular activity of SOD, decreased intracellular levels of catalase, elevated levels of xanthine oxidase and TBARS, and severely impaired singlet oxygen trapping were observed when compared to normozoospermic ejaculates. Interrelationships between SOD and TBARS, and between xanthine oxidase and catalase, appeared to be of key importance when analysed separately in seminal plasma and in spermatozoa or in a combination of both. Elevated xanthine oxidase levels and low capacity for singlet oxygen trapping are statistically significant factors for the evaluation of male infertility which can develop as a result of persistent oxidative stress.
The most common cause of male infertility is idiopathic. Standard investigations reveal no abnormality in such cases. The aim of the study was to investigate the levels of sperm DNA damage and seminal oxidative stress and their relationships with idiopathic infertility. The study included 30 normozoospermic infertile men seeking infertility treatment and 20 fertile donors. Semen analysis was performed according to the World Health Organization guidelines. Sperm DNA damage was assessed by alkaline single cell gel electrophoresis (comet assay) after preparation with two-step discontinuous Percoll gradient. Seminal oxidative stress was measured by a novel automated method. DNA damage score, total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI) were not different in idiopathic infertile men compared with controls. No correlations were also found between DNA damage score and TAS, TOS levels and OSI in idiopathic infertile group. We did not find any relationship between sperm DNA damage and oxidative stress in normozoospermic infertile men. We think that the pathophysiology of idiopathic infertility cannot be explained by sperm DNA damage or seminal oxidative stress.
Cryptorchidism and oligozoospermia are clinical conditions closely associated with impaired fertility. Oxidative stress and related sperm DNA damage have been identified as significant causes of male infertility.
To determine the extent of sperm nuclear DNA damage in patients affected with idiopathic oligozoospermia or undescended testes and to examine its relationship with oxidative stress.
We studied 20 patients with idiopathic oligozoospermia and 18 with undescended testes (who previously underwent orchiopexy) and 25 normozoospermic healthy controls. All subjects underwent semen analysis. Sperm DNA damage was evaluated by the sperm chromatin structure assay/flow cytometry (SCSA-FCM) and by the dUTP-biotin nick end labeling (TUNEL) assay. Levels of reactive oxygen species (ROS) and total antioxidant capacity (TAC) were assessed by a chemiluminescence assay.
DFI (percentage of sperm with denatured DNA) values and percentage of TUNEL positive cells were significantly greater in patients with oligozoospermia (DFI: 28.8+/-5.6; TUNEL+: 26.9+/-3.0) or cryptorchidism (DFI: 26.4+/-10.1; TUNEL+: 29.1+/-3.9), compared with controls (DFI: 7.1+/-0.9; TUNEL+: 14.2+/-1.2). Similarly, both groups of patients had significantly higher (p<0.01) levels of ROS. TAC levels did not differ between control and patient groups, suggesting that the DNA damage occurs before spermiation.
Sperm DNA damage is significantly increased in men with idiopathic oligozoospermia and in cryptorchid subjects. The finding of increased ROS levels may indicate that seminal oxidative stress may be involved in the pathogenesis of sperm DNA damage in these patients.
The lipid composition of the sperm membrane has a significant effect on the functional characteristics of spermatozoa.
In the present study, fatty acid composition of spermatozoa and seminal plasma levels of free 15-F(2t)-Isoprostane and catalase were assayed in men with normozoospermia, asthenozoospermia, asthenoteratozoospermia, and oligoasthenoteratozoospermia.
In spermatozoa from asthenozoospermic men only oleic acid levels showed a significant difference from normozoospermic men. In spermatozoa from asthenoteratozoospermic and oligoasthenoteratozoospermic samples all of the tested fatty acids were significantly higher than those from normozoospermic samples. Seminal plasma levels of catalase were significantly lower in all patients while levels of free 15-F(2t)-Isoprostane were significantly higher in all patients compared with normozoospermic men.
Spermatozoa from pathological samples may have higher levels of polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), than spermatozoa from normozoospermic men. Therefore, damage induced by lipid peroxidation would be higher in spermatozoa from pathological samples than those from normozoospermic men.
Sperm oxidative stress and the effect of an oral vitamin E and selenium supplement on semen quality in infertile men
- Keskes-Ammar
Vasectomy reversal associated with increased reactive oxygen species production by seminal fluid leukocytes and sperm
- Shapiro