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Phytoestrogens derived from red clover: An alternative to estrogen replacement therapy?

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Abstract

The benefits of plant extracts from soy and red clover as alternatives to conventional hormone replacement therapy (HRT) have been debated in the past. Here, an attempt has been made to summarize the biochemical and pharmacological data in the light of clinical aspects. Red clover and soy extracts contain isoflavones, which have a high affinity to estrogen receptor alpha (ERalpha), estrogen receptor beta (ERbeta), progesterone receptor (PR) and androgen receptor (AR). The higher affinity to ERbeta compared to ERalpha has been used as an explanation why red clover extracts function as food additives to treat menopausal disorders and may reduce risk of breast cancer. Biochemical analysis shows that these representatives of phytoestrogens have multiple actions beside selective estrogen receptor modulator (SERM)-activity. They act as selective estrogen enzyme modulators (SEEMs), have antioxidant activity and interact with transcription factors such as NF-kappaB. Furthermore, it is indicated that they have protective effects on osteoporosis and the cardiovascular system. Currently 40-50mg of isoflavones (biochanin A, daidzein, formononetin and genistein) are recommended as daily dose. This recommendation is based on the daily intake of phytoestrogens in a traditional Japanese diet.

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... Therefore, targeted isoflavone supplementation has been identified as an alternative and effective antimicrobial growth promoter. After absorption, isoflavones have phytoestrogenic activity that have been shown to cause arterial vasodilation via nitric oxide synthase activation [17][18][19][20]. In a study by Aiken et al. [21], isoflavone supplementation was demonstrated to relax arteries and reduce resistance of blood flow when goats were challenged with endophyte-infected tall fescue (E+ TF) seed. ...
... The legume source-dependent response observed could be attributed to the variable concentration and composition of isoflavones present. Red clover and white clover both contain predominantly biochanin A and formononetin, with lower levels of genistein and daidzein present [19,25,26]. However, these clovers are also different in their total isoflavone content. ...
... Jia and colleagues [52] were not able to detect any vasoactive impacts of the isoflavones' formononetin or biochanin A in bovine mesenteric arteries and veins exposed to E+ TF seed extract in vitro. This is consistent with reports that the vaso-activity of isoflavone metabolites may be greater than the pure isoflavones themselves [19,53]. In the rumen, resident microorganisms demethylate formononetin to daidzein and biochanin A to genistein. ...
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Ergot alkaloids produced by a fungal endophyte that infects tall fescue (Lolium arundinaceum; (E+ TF) can induce constriction of the vasculature in ruminants, resulting in “fescue toxicosis”. Legumes contain isoflavones that have been demonstrated to prevent and reverse E+ TF vasoconstriction. Several legumes are conventionally utilized in ruminant production, but can vary in both isoflavone concentration and composition. A feeding study was conducted to determine if isoflavone supplementation via red clover (Trifolium pratense), white clover (Trifolium repens), or soybean (Glycine max) meal can alleviate vasoconstriction when wether goats were challenged with E+ TF seed. The basal diet was chopped grass hay ad libitum. Carotid luminal areas were obtained pre- and post-ruminal infusions of E+ TF seed (15 µg kg BW−1 ergovaline + ergovalanine ± red clover, white clover, or soybean meal at 2.61 mg kg BW−1). When goats were challenged with E+ TF seed, the mean carotid luminal areas decreased by 56.1% (p < 0.01). All treatments were able to partially mitigate vasoconstriction, with red clover being the most effective (+39.8%), and white clover and soybean meal eliciting an intermediate response (+30%, p < 0.01). Results indicate that legumes can relax vasoconstriction in goats consuming ergot alkaloids, despite differences in isoflavone profile and concentrations.
... Пре-и постменопауза 2020; (13):99-104 ВВЕДЕНИЕ В последнее время в цивилизованном обществе статус женщины претерпел существенные изменения. Возросла роль представительниц прекрасного пола в политической жизни страны, государственном и общественном управлении, во всех отраслях экономики, культуры и науки, образовании, здравоохранении и спорте. ...
... Примером такой добавки могут быть капсулы Феминал, содержащие 40 мг сухого экстракта красного клевера. Стоит отметить, что, согласно данным исследований, цимицифуга не оказывает воздействие на эстрогеновые рецепторы [13,14]. ...
... Кроме того, все четыре изофлавоновые субстанции обладают выраженным аффинитетом к эстрогеновым рецепторам β (ER-β), что может лежать в основе тканеспецифичности экстракта красного клевера [15]. Более высокое сродство к ER-β по сравнению с ER-α исполь- зуется в качестве объяснения, почему экстракты красного клевера могут снижать риск рака молочной железы [13]. В этом же исследовании V. Beck биохимический анализ подтвердил, что изофлавоны красного клевера обладают действием селективных модуляторов эстрогеновых рецепторов (SERM) и селективных модуляторов активности стероидных энзимов (SEEM), проявляют антиоксидантную активность и взаимодействуют с факторами транскрипции (NF-γ B). ...
Article
In the civilized world, there is a dilemma on how to relate to aging - to perceive it as a natural process characteristic of any living organism or to elevate it to the rank of a disease in order to try to develop methods of “treatment”. Age-related changes in a woman begin at about 40 years old and are associated with various hormonal changes. Often this stage in the life of the fair sex does not go unnoticed, it makes unpleasant corrections to the lifestyle with the need to change established habits and rules. Interestingly, in the era of information technology, women are not always sufficiently informed about the main age-related changes in their own body. The primary task of clinicians is to timely reveal the secrets of women’s health and help them adapt to the upcoming changes. Fluctuations in hormonal status are observed throughout a woman’s life. Contrary to the common misconception among women, it is completely incorrect to identify menopause with old age, because the hormonal function in women begins to fade long before the true aging. Climax is a multistage period of life, during which all systems of the female body adapt to the new conditions of estrogen deficiency.In the treatment of climacteric disorders hormonal therapy with synthetic analogues of female sex hormones is used. Only preparations of bioidentical hormones have a real protective effect on metabolic disorders, osteoporosis, cardiovascular complications, cognitive disorders. According to clinical recommendations, approaches to the treatment of menopausal disorders should be clearly customized to the identified risks and expected benefits. Choosing the optimal composition and regime of MHT allows to increase efficiency and minimize possible risks. Clinicians should not forget about situations where there are restrictions or contraindications to the use of MHT. The presence of absolute and relative contraindications to MHT, as well as widespread hormoneophobia in our country in women have led to the development of alternative methods of treatment of menopausal disorders.
... Isoflavones found in red clover, are phytoestrogens, and possess anticancer properties. The isoflavones, formononetin and biochanin A, are both absorbed into the human body, then transformed to daidzein and genistein, respectively [3]. According to a lab technique called "chorioallantoic membrane assay", the results show that daidzein, genistein, and biochanin A, contribute to red clover's antiangiogenic and anti-inflammatory properties. ...
... According to a lab technique called "chorioallantoic membrane assay", the results show that daidzein, genistein, and biochanin A, contribute to red clover's antiangiogenic and anti-inflammatory properties. Biochanin A also inhibits aromatase activity and expression [3]. The greatest contributor to red clover's anticancer properties is the isoflavone formononetin, which has been extensively studied over the past decade. ...
... Red clover was studied as a complement to hormone replacement therapy. This extract contains isoflavones, mainly formononetin, which functions in treating menopausal disorders and reducing the risk of ovarian cancer due to high antioxidant properties [3]. ...
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The cytotoxic effects of red clover, blue false indigo, and red raspberry plant extracts were evaluated on CHO-K1 cells. The hormone-dependent CHO-K1 cells are ovarian cells derived from Chinese hamsters. Trifolium pratense (red clover) extracts were obtained from the blossoms and leaves of the red clover plant. Baptisia australis (blue false indigo) extracts were obtained from the roots, stems, and leaves of the blue false indigo plant. Rubus idaeus (red raspberry) extracts were prepared from the fruits of the red raspberry plant. Two methods, soxhlet and microwave assisted extractions, were utilized and evaluated for their effectiveness in producing phyto extracts. Methanol was the solvent used in both methods. In all experiments, the CHO-K1 cell line was exposed to the different extracts for a period of 48 hours. An MTS assay was performed to evaluate the effectiveness of the cytotoxic capabilities of each extract at different concentrations on the CHO-K1 cell line. After the collection of all the data, the 1:1 combination of red clover and red raspberry extracts, obtained via soxhlet extraction, yielded the most potent cytotoxic effects on the CHO-K1 cell line.
... Phytoestrogens are natural compounds structurally similar to E2 and may act as estrogen receptor (ER) agonists or antagonists [6]. They are frequently used in phytomedicine to treat menopausal symptoms and disorders such as osteoporosis, cardiovascular disease and breast cancer [7]. ...
... BCA has drawn considerable attention due to its wide spectrum of pharmacological activities, including neuroprotective, anti-cancer, antioxidant, anti-inflammatory, anti-allergic and anti-hyperglycemic effects [10]. A previous study showed that BCA-treated OVX mice had decreased bone loss and unchanged uterine weight, while an increase in uterine weight was observed in the E2-treated group [11,12], implying that phytoestrogens might be an alternative to E2 replacement therapy and that BCA may alleviate arthritis [6,13]. ...
Article
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Objective and design: Biochanin A (BCA), a phytoestrogen, has various pharmacological properties. This study was conducted to compare BCA's therapeutic property against 17-β estradiol replacement therapy in zymosan-induced arthritis (ZIA) in mice. Additionally, we further investigated in vitro the anti-inflammatory action on neutrophils. Treatment: Ovariectomized (OVX) and non-OVX mice were pretreated with BCA (1, 3 and 9 mg/kg) or estrogen (50 µg/kg) for 14 days prior to ZIA. Neutrophils were pretreated with BCA (1, 10 and 100 μM) for 1 h prior to phorbol 12-myristate 13-acetate. Methods: Anti-inflammatory effects of BCA were evaluated by cellular infiltrate, paw edema and cytokine measurement. In vitro, apoptosis was assessed by morphology and flow cytometry. Neutrophil extracellular traps (NET) were determined by fluorescent microscopy and DNA release. Statistical differences were determined by one- or two-way ANOVA. Results: BCA inhibited neutrophil accumulation, paw edema and proinflammatory cytokine (TNF-α and IFN-γ) and increased anti-inflammatory cytokines (IL-4 and IL-10) in OVX and non-OVX mice, similar to 17-β estradiol replacement therapy. In vitro, BCA increased apoptosis and consequently reduced NETs. Conclusion: BCA has a notable anti-inflammatory effect, similar to 17-β estradiol, and is especially effective for treatment of ZIA. These results suggest that BCA may be promising for the treatment of postmenopausal arthritis.
... Due to their estrogenic effect, isoflavones have been proposed as an alternative to hormone replacement therapy in postmenopausal women [112,113]. The reported effects range from decreased bone loss [114] to reduced incidence of hot flushes [115][116][117]. ...
... Dietary supplements based on red clover extracts are readily available in the market with different isoflavone composition and quantity [112,115,137]. The supplements typically contain 40-80 mg isoflavones per capsule, which agrees with the recommended daily doses of 40-50 mg/day [113] or 40-82 mg/day [116]. ...
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In a circular economy, products are made from renewable resources and the waste streams generated during production are either reused, recycled or recovered. The Biocascade methodology considers bio-waste as a resource that can be exploited to produce high-value products such as pharmaceuticals, food ingredients and nutrients; and low-value products such as feed, energy or soil conditioners. The Biocascade principle ensures optimal biomass exploitation by following a hierarchy from high-to-low value, where the waste from one process is the starting material for the next. Biowaste from plant origin is a very suitable resource for applying the Biocascade methodology, both in terms of worldwide production and of variety of components. In this review, the biowaste from sour cherry wine, ornamental kalanchoe plants and red clover feed production, have been examined for processing using a Biocascade approach. Starting from the biowaste characterization, the most relevant components have been identified highlighting their potential uses. The extraction methodology is then discussed in terms of solvent used, operating conditions and yield. Based on the information retrieved from literature, different process flowsheets have been proposed to maximize the use of the biowaste following the Biocascade perspective and targeting zero-waste generation. Please read the full article here: https://link.springer.com/epdf/10.1007/s12649-020-01082-6?sharing_token=c63munoNjBKieEHNmu4GBve4RwlQNchNByi7wbcMAY5fCE8UPEkanRT_Bqls9i1BfcVBvA4U81WNu0f2bCi7p6mmJQIFF9vP7TxocIP563qJLIx9Ud4LDxojQLaCnePfjuLoV8IC6ZwcgnWNbJx712J6j0alA_yRod4Nvkq9KJg%3D
... Phytoestrogenic compounds found in red and white clo- ver have been reported to improve overall growth performance of livestock grazing endophyte-infected tall fescue throughout the spring and summer months [11][12][13]. Var- ious isoflavones have been targeted for use in human medicine due to their beneficial reduction in breast cancer symptoms, improvements in cardiovascular health, as well as reductions in menopausal symptoms [14][15][16]. As such, determining additional benefits of isoflavones, such as the improvement of fiber utilization and rumen fermentation in livestock, is timely. ...
Article
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Negative impacts of endophyte-infected Lolium arundinaceum (Darbyshire) (tall fescue) are responsible for over $2 billion in losses to livestock producers annually. While the influence of endophyte-infected tall fescue has been studied for decades, mitigation methods have not been clearly elucidated. Isoflavones found in Trifolium pratense (red clover) have been the subject of recent research regarding tall fescue toxicosis mitigation. Therefore, the aim of this study was to determine the effect of ergovaline and red clover isoflavones on rumen microbial populations, fiber degradation, and volatile fatty acids (VFA) in an in vitro system. Using a dose of 1.10 mg×L-1, endophyte-infected or endophyte-free tall fescue seed was added to ANKOM fiber bags with or without 2.19 mg of isoflavones in the form of a control, powder, or pulverized tablet, resulting in a 2×3 factorial arrangements of treatments. Measurements of pH, VFA, bacterial taxa, as well as the disappearance of neutral detergent fiber (aNDF), acid detergent fiber (ADF), and crude protein (CP) were taken after 48 h of incubation. aNDF disappearance values were significantly altered by seed type (P = 0.003) and isoflavone treatment (P = 0.005), and ADF disappearance values were significantly different in a seed×isoflavone treatment interaction (P�0.05). A seed×isoflavone treatment interaction was also observed with respect to CP disappearance (P�0.05). Eighteen bacterial taxa were significantly altered by seed×isoflavone treatment interaction groups (P� 0.05), eight bacterial taxa were increased by isoflavones (P�0.05), and ten bacterial taxa were altered by seed type (P�0.05). Due to the beneficial effect of isoflavones on tall fescue seed fiber degradation, these compounds may be viable options for mitigating fescue toxicosis. Further research should be conducted to determine physiological implications as well as microbiological changes in vivo.
... In recent years, the scientific consideration in phytoestrogens has grown significantly, especially in the medical field [8,9]. These plant combinations are comparable to animal estrogens [10], which are commonly used in aquaculture, and they also have excessive costs compared to plant extracts and their use requires expert labor. ...
Article
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The main purpose of this study was to consider the impact of Vitex agnus-castus hydroalcoholic extract, containing phytoestrogenic compounds on growth indices, sex ratio and histology of gonads of female Zebrafish. Fish larvae (4-day-old after hatching) were nourished with investigational diets (0 (control group T0), 5 g (T1), 10 g (T2) and 15 g (T3) kg −1 food) from first active feeding for 90 days. The results showed that Condition Factor (CF), Food Conversion Rate (FCR), Body Weight Gain (BWG), and Specific Growth Rate (SGR) were affected meaningfully by hydroalcoholic extract of Vitex agnus-castus (p < 0.05). These factors were significantly higher in compare to T3 treatment (p < 0.05). With increasing concentration of the extract in the diet, the ratio of female to male increased Treatment of T3 resulted in 87.23% feminization. Overall, this study suggests the use of T3 treatment achieves the best reproduction performance and a higher percentage of females in zebrafish as a model for aquaculture species. According to the results, the lowest levels of glucose and cholesterol were observed in T3 treatment and showed a significant changes with the control and treatments (p < 0.05).
... Moreover, the phytochemical compounds of red clover such as genistein and daidzein could interfere with steroid formation in rat adrenal and consequence decreased testosterone levels (34,35). In addition, coumestrol, biochanin, phytoestrogens and zearalenol have inhibitory effects on steroidogenic enzyme activity of 17 betahydroxysteroid dehydrogenases (17β-HSD) which result in reduced androgens synthesis (36). ...
Article
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Polycystic ovary syndrome (PCOS) is considered as one of the leading endocrine disorders during reproductive age in women. This study designed to determine the therapeutic effects of red clover (Trifolium pratense) on letrozole-induced PCOS in vivo. Forty female Sprague-Dawley rats were equally divided into five groups. Control group with a regular sexual cycle received normal saline (letrozole vehicle). Letrozole (1 mg/kg) was used to induce the PCOS to the rats in the treatment groups. After induction of PCOS, four treatment groups received the normal saline, or clomiphene citrate (1 mg/kg), or red clover extracts (500 or 750 mg/kg) for 30-days. After treatment, ovary and uterus were removed, weighed, and the ovaries were subjected to histopathological studies. Serum testosterone and estradiol levels, antioxidant activities, and lipid profiles were evaluated. Red clover extracts and clomiphene citrate decreased testosterone levels and showed a significant increase in estradiol levels in comparison to PCOS induced group (p<0.05). Red clover administration restored the GSH, SOD and CAT levels (p<0.05) and decreased the NO and MDA levels (p<0.05). Treatments caused no significant change in levels of TG, TC, and FBG factors when compared to PCOS induced group (P>0.05). However, red clover (750 mg/kg) significantly increased HDL and decreased LDL levels when compared to PCOS induced group (P<0.05). Treatment with red clover reduced ovarian weight, volumes of ovarian, medulla, cortex and number of cysts and increased number of oocytes compared to PCOS group. Both red clover and clomiphene citrate could treat the letrozole induced PCOS in rats; however, red clover indicated antioxidant activities more than clomiphene citrate. Red clover may be used for discovering anti-PCOS drugs with lower side effects.
... However, no clear evidence for a breast cancer preventive effect o f a phytoestrogen-rich diet has been found and some laboratory findings even indicate stimulatory effects o f phytoestrogens on breast cancer development (Kortenkamp, 2007a). An increase in the bone mineral density, and therefore a reduced risk o f developing osteoporosis and cardioprotective activity, are other reported beneficial effects o f the phytoestrogens, which were found in menopausal women (Beck et al, 2005). ...
Thesis
The life of most women is at some point affected by side effects of their menopausal transition. The most common menopausal side effects are the hot flushes, which influence the quality of life severely. The physiology behind the hot flushes and the connection between hormones, neurotransmitters and hot flush occurrence is still unclear. However, in vivo studies suggest a correlation between the hormone 17-β-estradiol (E2), the neurotransmitter serotonin (5-HT) and the regulation of the core body temperature. Menosan® (a Salvia officinalis tincture) is an herbal medicinal product (HMP), which has proven to reduce hot flushes, but the mechanism underlying this effect has remained unknown. In order to elucidate the anti-hot flush mechanism of action, the Salvia officinalis tincture as well as three subextracts (n-hexane, chloroform and aqueous-ethanolic (aq-EtOH) subextract (SE)), which were obtained by solvent-solvent partitioning of the tincture, were tested for relevant in vitro activities. In a multi-target approach, the influence of the tincture and SE on the serotonergic system was assessed in a selective serotonin re-uptake inhibition (SSRI) assay and estrogenicity was investigated in an estrogen receptor luciferase reporter gene (ERLUX) assay and an estrogen screen (ESCREEN) assay. None of the extracts showed activity in the SSRI assay. The aq-EtOH SE was found to be estrogenic in the ERLUX assay (EC50 64 μg/ml) and the ESCREEN assay (EC50 130 μg/ml), but none of the other extract exhibited an estrogenic response. In a second approach the Salvia officinalis tincture was investigated for a possible multi-beneficial effect for menopausal women and whether the active principles underlying the anti-hot flush activity and treatment of Alzheimer disease could be linked. The Salvia officinalis tincture and SEs were therefore tested for acetylcholinesterase inhibition (AChEI) activity. The neurotransmitter acetylcholine (ACh) has been linked with Alzheimer diseases (AD) and as it has been suggested that menopausal women may have a higher risk than men to develop AD, hence the effect on ACh levels would be of interest. No activity in the AChEI assay was found for the Salvia officinalis tincture or the SEs. HMPs need to meet high safety standards. Thus, for the safety assessment the inhibitory potential of the Salvia officinalis tincture and its SEs against CYP3A4 was evaluated in an in vitro assay. Cytochrome P450 (CYP) enzyme 3A4 is as key enzyme in human drug metabolism and all four extracts inhibited the CYP3A4 enzyme. While the activity of the aq-EtOH SE was very low, the activity of the Salvia officinalis tincture (EC50 6.8 μg/ml), the chloroform SE (EC50 3.1 μg/ml) and the n-hexane SE (EC50 1.8 μg/ml) were considerably stronger and the inhibitory principle of these extracts warrant further investigation. In the initial screening the aq-EtOH SE was the only SE, which displayed an anti-hot flush activity. Hence, an activity-guided fractionation of the aq-EtOH SE was performed in order to isolate and identify the active constituent(s). Following the estrogenicity in the ERLUX assay resulted in identification of luteolin-7-O-β-glucuronide as the active principle in fraction AQ 4. The estrogenic potential of the isolated luteolin-7-O-β-glucuronide was weak (EC50 129 μg/ml). Luteolin-7-O-glucoside was identified as the major component in the most active fraction (AQ 7.6.7.6, EC50 0.7 μg/ml). The compounds were identified in the estrogenic fractions by nuclear magnetic resonance (NMR) spectroscopy, mass spectroscopy (MS), ultra-high-pressure liquid chromatography at elevated temperature coupled to time-of-flight mass spectrometry (UHPLC-TOF-MS) and capillary (cap) NMR. The estrogenic activity of pure commercially available luteolin-7-O-glucoside (EC50 0.44 μg/ml) was similar to that of fraction AQ 7.6.7.6. Quantitative analysis of luteolin-7-O-glucoside in the aq-EtOH SE and the Salvia officinalis tincture explained the estrogenic activity of the aq-EtOH SE, but failed to explain the absence of estrogenic effects by the Salvia officinalis tincture. In conclusion the investigation revealed noteworthy CYP3A4 inhibitory activity by the Salvia officinalis tincture, luteolin-7-O-β-glucuronide has been identified as the estrogenic principle in fraction AQ 4 and luteolin-7-O-glucoside as the putative estrogenic principle of fraction AQ 7.6.7.6.
... Menopause is a natural process that develops in women typically in their late 40s and involves a decrease in the estrogen levels, particularly due to ovarian decline [1,2]. is decrease in estrogen levels is accompanied by various symptoms, such as hot flashes, night sweats, and depression [3]. Estrogen deficiency can lead to an increased risk of metabolic diseases, such as obesity, heart disease, diabetes, and hypertension [4][5][6]. Furthermore, it affects the atherogenic lipid profile, such as the levels of high-density-lipoprotein (HDL) and low-density-lipoprotein (LDL) cholesterol [7]. ...
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Red clover and hops are already known for their alternative menopausal therapies; however, their combination has not yet been studied. This study aimed to evaluate the efficacy of the combination of red clover and hops extract (RHEC) for treating menopausal symptoms for the first time. A high-performance liquid chromatography (HPLC) method for RHEC was developed and validated for the analysis of biochanin A in red clover extract and xanthohumol in hops extract. An in vivo study was conducted using an ovariectomized rat model treated with RHEC (125, 250, and 500 mg/kg, p.o.) for a 12-week test period. Changes in body weight, tail skin temperature (TST), serum lipid profile, bone metabolism, antioxidants, and markers of vasorelaxation and uterus endometrium were evaluated. RHEC significantly inhibited body weight gain and decreased fat weight. Changes in TST associated with flashes were significantly inhibited in the RHEC groups. Other markers related to menopausal symptoms, such as blood lipid profile (total cholesterol and low-density-lipoprotein cholesterol), bone metabolism (serum alkaline phosphatase, osteocalcin, and c-terminal telopeptide type 1), antioxidants (superoxide dismutase and malondialdehyde), and vasorelaxants (endothelin-1 and nitric oxide), were significantly improved after the administration of RHEC. We also confirmed the safety of RHEC through histopathological observation of the endometrium. Our findings demonstrate that RHEC appears to have high potential for comprehensively improving various symptoms of menopause.
... According to the quantitative analysis, the sum of the isoflavones with estrogenic activity (daidzein, genistein, glycosides, apigenin, formononetin, myriceitin, and glycitein) was 906.1 µg/g DW for red clover and 587.5 µg/g DW for alfalfa sprouts on the third day of germination. Due to the different compounds' bioavailability [43], consideration of their biological activity could be speculative. ...
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Identification and quantification of polyphenols in plant material are of great interest since they make a significant contribution to its total bioactivity. In the present study, an UPLC-Orbitrap-MS/MS approach using the variable data acquisition mode (vDIA) was developed and applied for rapid separation, identification, and quantification of the main polyphenolic compounds in Medicago sativa L. and Trifolium pratense L. sprouts in different germination stages. Based on accurate MS data and fragment ions identification strategy, a total of 29 compounds were identified by comparing their accurate masses, fragment ions, retention times, and literatures. Additionally, a number of 30 compounds were quantified by comparing to the reference standards. Data were statistically analysed. For both plant species, the sprouts of the third germination day are valuable sources of bioactive compounds and could be used in phytotherapy and nutrition. Although Trifolium pratense L. (Red Clover) is considered to be a reference for natural remedies in relieving menopause disorders, alfalfa also showed a high level of biological active compounds with estrogenic activity.
... Soybean and soybean derived products have recently become common additives in health foods and beverages (7,8). Soybeans have able to bind and activate estrogen receptor (9). Particularly, the binding affinity of estrogen receptor B (ERB) for the isoflavonoid genistein is 87% that for estradiol, and genistein activates ERB with high potency (10). ...
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Abstract The present study was conducted to investigate the effect of soybean seeds (15g /kg diet) for one month on blood picture, total cholesterol(T-C), high density lipoproteins(HDL), low density lipoproteins(LDL), triglycerides (TG), glucose, alanine aminotrasferase (ALT) and aspartate aminotrasferase (AST) enzymes activity, glutathione(GSH), total proteins, albumin, globulin and body weight in ovariectomized rats(OVX). The rats randomly divided in to three groups (6 rats / group). The first group served as a control, the second group rats at one month aged bilateral ovariectomy were done, and the third group rats at one month aged bilateral ovariectomy were done, and after two months the rats treated with soybean seeds for one month. The result showed that a significant decrease in the total leucocytes count, HDL, GSH and globulin in ovariectomized group compared with control group, with no significant differences in PCV, Hb, differential leucocytes count(DLC), total proteins, albumin and body weight. The current study demonstrated that a significant increase in T-C, LDL, ALT and glucose in ovariectomized rats group compared with control group. Administration of soybean seeds caused a significant increase in total leucocytes count, globulin and body weight compared with ovariectomized group, and a significant decrease in T-C, TG, LDL and glucose compared with ovariectomized group. It could be concluded from this study that administration of soya beans seeds have a beneficial improving of some physiological and biochemical parameters in ovarictomized rats.
... DHD has important functions, apart from its role as a precursor for equol [11][12]. Due to similarities in the structure of DHD and 17β-estradiol, DHD can effectively reduce vasoconstriction, so DHD is one of the active ingredients that maintain the activity of blood vessels to prevent vascular endothelial injury [13][14]. Atkinson et al. found that some antibiotics inhibit the production of equol but have no effect on DHD production [15]. ...
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The aim of this research was to isolate bacteria capable of biotransforming daidzein from fresh feces from pregnant horses. A Hungate anaerobic roller tube was used for anaerobic culture. Single colonies were picked at random and incubated with daidzein. High performance liquid chromatography was used to detect whether the isolated bacteria were able to biotransform the substrate. A strain capable of reducing daidzein was selected and characterized using sequence analysis of 16S rDNA, and a phylogenetic tree was constructed. The morphological physiological and biochemical characteristics of the strain were investigated. A facultative anaerobic, Gram-positive bacterium capable of converting daidzein to dihydrodaidzein was isolated and named HXBM408 (MF992210). A BLAST search of HXBM408's 16S rDNA sequence against the GenBank database suggested that the strain has 99% similarity with Pediococcus acidilactici strain DSM (NR042057). The morphological, physiological, and biochemical characteristics of HXBM408 are very similar to those of Pediococcus. Based on these characteristics, the strain was identified as Pediococcus acidilactici. The bacterial strain HXBM408 isolated from the feces of pregnant horses was able to reduce the isoflavone daidzein to dihydrodaidzein.
... Currently, various parts of this medicinal plant are used in the management and treatment of many disorders, specifically digestive system disorders. Moreover, it is highly useful in the treatment of inflammation of bronchioles, chronic cough, diabetes mellitus, kidney stones as well as in nausea and vomiting [87]. ...
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Polycystic Ovarian Syndrome (PCOS) is multi-organ syndrome that affects 6-25% of females during reproductive age. It disrupts normal hormone levels of females and affects adrenal hormone and sex hormones along with pituitary hormones including adrenocorticotropic, growth hormone and gonadotropins hormones. It results in several secondary characteristics in females such as infertility , hormonal imbalance, oligomenorrhea, amenorrhea, obesity and hirsutism. Polycystic ovarian syndrome is associated with mental and reproductive disorder and almost 4-18% mature female students are affected by polycystic ovarian syndrome. Female affected by polycystic ovarian syndrome have increased risk of infertility, ovarian cancer, skin problems and psychological problems such as anxiety, depression and changes in sleep along with mood swings. This chapter discusses the Prevalence, Symptomatology and Management of Polycystic Ovarian Syndrome. For the management of PCOS, the role of some medicinal plants including Asparagus racemosus, Tinospora cordifolia, Foeniculum vulgare, Ocimum tenuiflo-rum, Actaea racemosa, and Lepidium meyenii have also been discussed in addition to other treatment modalities. The literature data was searched out and compiled using relevant original research articles and reviews published and indexed in Google Scholar, Scientific Information Database, Elsevier, PubMed and Science Direct.
... Recently, medical research focusing on BCA has increased because of its various purported biological activities, including its antioxidant, anti-inflammatory, anti-infective, and anticarcinogenic effects, and BCA has been used for several purposes, such as to treat estrogen deficiency and pain and reduce the severity of nerve damage (Puli et al., 2006;Medjakovic and Jungbauer, 2008). This extract from plants is already commercially available because of its potential benefits to human health and because it is considered innocuous (Howes et al., 2002;Atkinson et al., 2004;Beck et al., 2005;Sklenickova et al., 2010). Most commercial products are composed of several isoflavone contents, including BCA (Booth et al., 2006;Ahmad et al., 2013). ...
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Biochanin A (BCA) is an isoflavone mainly found in red clover with poor solubility and oral absorption that is known to have various effects, including anti-inflammatory, estrogen-like, and glucose and lipid metabolism modulatory activity, as well as cancer preventive, neuroprotective, and drug interaction effects. BCA is already commercially available and is among the main ingredients in many types of supplements used to alleviate postmenopausal symptoms in women. The activity of BCA has not been adequately evaluated in humans. However, the results of many in vitro and in vivo studies investigating the potential health benefits of BCA are available, and the complex mechanisms by which BCA modulates transcription, apoptosis, metabolism, and immune responses have been revealed. Many efforts have been exerted to improve the poor bioavailability of BCA, and very promising results have been reported. This review focuses on the major effects of BCA and its possible molecular targets, potential uses, and limitations in health maintenance and treatment.
... Therefore, perimenopausal women choose to seek alternative therapies from some functional plant extracts. Phytoestrogens are similar with mammalian estrogens on structurally and functionally, but with less side effects compared with synthetic HRT [8,9]. Phytoestrogens have been considered as selective estrogen receptor modulators AGING (SERMs), it could bind to estrogen receptors (ERs) and appear to have various estrogenic and antiestrogenic effects [10,11]. ...
Article
The reproductive tissues are negatively influenced by estrogens in hormone therapy. Qingyan formula ethanol extract (QYFE)'s estrogenic effects and safety on reproductive tissues after long-term administration and its mechanism via estrogen receptor (ER) pathway haven't been studied. Here, we characterized its estrogenic effects using ovariectomized rats together with in vitro studies for further molecular characterization. Ovariectomized rats were treated with QYFE at doses of 0.7, 1.4, and 2.8g/kg for 12 weeks. The results showed QYFE has a potent estrogenic activity, as indicated by restoring the disappeared estrous cycle, antagonizing the atrophy of uterus, vagina and mammary gland, and the estrogen decline in circulation caused by ovariectomy. In addition, QYFE upregulated ERα and ERβ expressions and had a less stimulatory effect on PCNA and ki-67 antigen in reproductive tissues compared with estradiol valerate. QYFE components can bind to ERα and ERβ, significantly increased ERα/β-ERE luciferase reporter gene expression, upregulated the expressions of ERs, PR and pS2 in MCF-7 cells at protein and gene level. All these activities were significantly inhibited by the ER antagonist ICI182,780. QYFE's estrogenic activity maybe mediated by stimulating biosynthesis of estrogen and increasing the quantity of ERs in target tissue and via active ER to ERE-independent gene regulation.
... A traditionally well-accepted option comes from natural products. For example, the prolonged use of soybean products 8 and several preparations of Curcuma longa L., 9 Trifolium pratense L., 10,11 and Cimicifuga racemosa L. 12 has presented very interesting data. Despite these benefits, however, some caffeine-rich medicinal specimens, including Coffea arabica L., 13 may increase the risk of fractures in osteoporosis patients. ...
Article
Osteoporosis is a systemic bone disease that is characterized by impairments in bone strength that predispose an individual to a higher risk of fractures. Despite the various etiologies, undoubtedly the most important factors are aging of the population and hypogonadism. Although several therapeutic options are available, pharmacological treatments have some risks. Among these are increases in the incidence of thrombosis, breast cancer, ovarian cancer, endometrial cancer, and muscle injury, among others. Herbal medication may be an alternative for the treatment of osteoporosis. Thus, the aim of this study was to evaluate the therapeutic effect of a standardized extract of Tribulus terrestris L. (TT) on ovariectomy (OVX)-induced bone loss in rats. Female rats were first subjected to OVX and treated with TT (3, 30, and 300 mg/[kg·day]) or furosemide (25 mg/kg) orally for 28 days. Bone densitometry and tibial histology were performed, and acute renal function and testosterone, dehydroepiandrosterone (DHEA), and estradiol levels were assessed. Prolonged treatment with TT stimulated bone mass gain in all ovariectomized animals, raising bone mass to levels that were similar to sham-operated rats. DHEA levels significantly increased in TT-treated rats. The TT group also had lower calcium (Ca ²⁺ ) excretion that OVX control and furosemide-treated rats. Finally, the histopathological analyses showed the maintenance of bone turnover in all TT-treated groups. Overall, the results indicate that the standardized extract of T. terrestris exerted a bone-protective effect by increasing bone mineral density. This activity may be at least partially attributable to an increase in serum DHEA levels and a Ca ²⁺ -sparing effect.
... 10,11 Among the herbs, sources of phytoestrogens are soybeans, fenugreek, red clover, and licorice. 12,13 Licorice is one of the plants containing herbal estrogen and is effective in the body's hormonal balance. Its estrogen is of steroidal type and is similar to estrone (E1) and estradiol (E2) which are produced in the body. ...
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Background Vaginal atrophy is one of the most common menopausal complications affecting women's quality of life. The present study aimed to investigate the impact of Glycyrrhiza glabra (licorice) vaginal cream on vaginal signs and symptoms of vaginal atrophy in postmenopausal women. Methods This randomized controlled trial was conducted on 70 menopausal women, referring to health centers of Izeh city, located in the south west of Iran, from May to November 2017. The participants with vaginal atrophy were randomly assigned into two groups of 35. One of the groups received licorice 2% vaginal cream while the other was given placebo over a period of 8 weeks. Data were analyzed using SPSS software at the significance level of 0.05. Results The results indicated that at the baseline, none of the subjects (0%) in either groups had a vaginal cells maturation Index within 65–100 in MVI category. However, after 8 weeks of therapy, it improved significantly to 82.9% in Licorice group and 11.4% in the placebo group (p < 0.001). Also, the vaginal mucus cells changed from the baseline cells to intermediate and superficial cells within and between the two groups after the treatment (p < 0.001). Finally, the pH level significantly decreased in licorice group over time (p < 0.001). Conclusion Therapeutic use of licorice vaginal cream can improve the signs and symptoms of vaginal atrophy in postmenopausal women. Accordingly, use of licorice as a natural vaginal cream in vaginal atrophy can be suggested.
... The richness of leaf isoflavones in subterranean clover confirms previous results on this species [28]. Red clover isoflavone extracts are commercially available as nutraceuticals and they have been proposed as an alternative to hormone-replacement therapy [53]. Subterranean clover may represent an interesting new source of isoflavones, with higher concentration of these compounds and more diverse pattern of isoflavone composition compared to red clover [28]. ...
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Phenolics are important mediators in plant-environment interactions. The presence and concentration of phenolic compounds and their antioxidant activity were evaluated in leaves and flowers of a set of Trifolium species originating from contrasting environments encompassing lowland and mountain sites. The current germplasm proved a great reservoir of phenolic compounds, with different chemical structure and, possibly, diversified biological activity. Germplasm groups with specific phenolic composition were observed. In some cases, different patterns bore a taxonomic meaning. Lowland germplasm showed higher concentration of total phenolics in leaves than mountain accessions (50.30 vs. 34.19 mg/g dry matter (DM)), while the latter had higher concentration in flowers (114.16 vs. 57.44 mg/g DM). Outstanding concentration of isoflavones was observed in leaves of lowland germplasm (24.19 mg/g DM), and of both proanthocyanidins and flavonoids in flowers of mountain germplasm (53.81 and 56.62 mg/g DM, respectively). The pattern of phenolic composition in lowland and mountain germplasm was suggestive of different adaptive strategies. Three assays of antioxidant activity were tested, which were characterised by rather different reactivity towards phenolic composition. The scavenging activity was higher for leaf extracts of lowland germplasm, and for flower extracts of mountain germplasm. Besides identifying germplasm of interest, this study also suggested possible links between environmental factors and concentration and composition of phenolic compounds.
... Moreover, other minor aglycones, such are glycitein, irilone, orobol, pratensein, pseudobaptigenin and prunetin, as well as minor malonyl glycosides can be found (Spanguolo et al. 2014). Isoflavones are usually present in plants as glycosides and glycoside malonates: inactive forms that are suitable for storage of less soluble flavonoid aglycones (Beck et al. 2005;Xiao et al. 2017). It has also been reported that isoflavone glycosides are prone to hydrolysis by the enzyme β-glucosidase, which can be activated upon mechanical damage of plant tissue (Xiao et al. 2017). ...
Article
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This paper summarizes published literature concerning different factors that could affect the content and composition of isoflavones in red clover (Trifolium pratense L., Fabaceae). Influence of genotype, plant part, geographic origin, ontogeny and storage conditions of raw material are discussed. Statistical analysis of available data on the profile of isoflavones in red clover suggest that, in general, samples are grouped according to country of origin, indicating the importance of climate conditions. An increase in biosynthesis of isoflavones as a response to different abiotic factors is also considered. Knowledge about these factors is of great practical importance, since red clover is a medicinal plant widely used for production of dietary supplements, and producers aim to breed cultivars with high isoflavone concentrations.
... Therefore, phytoestrogens are important to be used as alternatives. Both structurally and functionally, phytoestrogens are similar to mammalian estrogens but with lower side effects compared with synthetic HRT reportedly (Beck, Rohr, & Jungbauer, 2005;Benassayag, Perrot-Applanat, & Ferre, 2002). Traditional medicines (TM) have been used for centuries in China for treatment of perimen- opausal syndrome and have attracted the attention of researchers *Equal contribution regarding TM as new phytoestrogens resource. ...
Article
Radix Paeoniae Alba (RPA) is widely used in clinical treatment for gynecological diseases, particularly abnormal menstruation, menstrual pain, and breast tenderness; however, no scientific evidence base links RPA to estrogen replacement therapy. In this study, we characterize estrogenic activity of RPA using immature and ovariectomized (OVX) mice together with in vitro studies focus on estrogen receptor (ER) pathway for molecular mechanism. RPA treatments demonstrated significant estrogenic activity, as indicated by promoting the development of uterus and vagina in immature mice, reversing the atrophy of uterus and vagina in OVX mice, up‐regulating the expressions of ERα and ERβ at protein and mRNA level in reproductive tissues. Meanwhile, RPA significantly increased serum estradiol and clearly decreased serum luteinizing hormone and follicle‐stimulating hormone of immature/OVX mice. Moreover, RPA could induce ER positive MCF‐7 cell from S‐phase to G2 stage and induce proliferation and no influence on ER negative MDA‐MB‐231 cell. RPA could bind with ERα and ERβ and significantly stimulate ERα/β‐estrogen response element (ERE) luciferase reporter gene expression. All activities were inhibited by the ER antagonist ICI 182,780. This study illustrates RPA exerts estrogenic effects by stimulating biosynthesis of estrogen in circulation, up‐regulating ERs in target tissues, and mimicking the estrogen through ER‐ERE‐dependent pathway.
... Currently, various parts of this medicinal plant are used in the management and treatment of many disorders, specifically digestive system disorders. Moreover, it is highly useful in the treatment of inflammation of bronchioles, chronic cough, diabetes mellitus, kidney stones as well as in nausea and vomiting [87]. ...
... animal models with acute and chronic inflammation demonstrated a marked anti-inflammatory and antioxidative activity of BA against organ injury (13). BA-mediated inhibition of bone loss in postmenopausal women has also been reported in a previous study (14). These findings suggested that BA treatment can potentially regulate bone cell proliferation, apoptosis and migration, leading to reduced bone loss and bone turnover. ...
Article
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Biochanin A (BA) is an organic compound produced by Trifolium pretense and Arachis hypogaea with anti-inflammatory and antioxidative effects. The aim of the current study was to evaluate the effects of BA on gingival inflammation and alveolar bone destruction in rats with experimental periodontitis. Experimental rats (n=25) were distributed equally into five groups: i) Healthy control (control) group; ii) experimental periodontitis (ligation) group; and iii) and ligation plus low, medium and high dose of BA (12.5, 25 and 50 mg/kg/day, respectively) groups. A nylon ligature was inserted around rats' maxillary molars for 14 days to trigger the experimental periodontitis. BA was intravenous injected once daily for 4 weeks. After that, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), reactive oxygen species (ROS) and osteocalcin (OCN) levels were determined in gingival and/or serum samples using ELISA or reverse transcription-quantitative PCR. Alveolar bone volume was assessed via hematoxylin and eosin staining and micro-computed tomography. Osteoclasts were identified by tartrate-resistant acid phosphatase staining, and the level of the nuclear factor erythroid-2 related factor 2 (Nrf2) was also detected by immunohistochemical staining. BA treatment groups showed alleviated alveolar bone resorption compared with the ligation group. Moreover, BA treatment significantly inhibited IL-1β, TNF-α, ROS levels, and reduced leukocyte acid phosphatase-positive cells, as well as increased OCN and Nrf2 levels compared with the ligation group. BA had beneficial effects on experimental periodontitis of rats. BA treatment inhibited inflammation, regulated unbalanced oxidative stress response and ameliorated the alveolar bone loss.
... Moreover, the phytochemical compounds of red clover such as genistein and daidzein could interfere with steroid formation in rat adrenal and consequence decreased testosterone levels (34,35). In addition, coumestrol, biochanin, phytoestrogens and zearalenol have inhibitory effects on steroidogenic enzyme activity of 17 betahydroxysteroid dehydrogenases (17β-HSD) which result in reduced androgens synthesis (36). ...
Article
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Polycystic ovary syndrome (PCOS) is considered as one of the leading endocrine disorders during reproductive age in women. This study designed to determine the therapeutic effects of red clover (Trifolium pratense) on letrozole-induced PCOS in vivo. Forty female Sprague-Dawley rats were equally divided into five groups. Control group with a regular sexual cycle received normal saline (letrozole vehicle). Letrozole (1 mg/kg) was used to induce the PCOS to the rats in the treatment groups. After induction of PCOS, four treatment groups received the normal saline, or clomiphene citrate (1 mg/kg), or red clover extracts (500 or 750 mg/kg) for 30-days. After treatment, ovary and uterus were removed, weighed, and the ovaries were subjected to histopathological studies. Serum testosterone and estradiol levels, antioxidant activities, and lipid profiles were evaluated. Red clover extracts and clomiphene citrate decreased testosterone levels and showed a significant increase in estradiol levels in comparison to PCOS induced group (p<0.05). Red clover administration restored the GSH, SOD and CAT levels (p<0.05) and decreased the NO and MDA levels (p<0.05). Treatments caused no significant change in levels of TG, TC, and FBG factors when compared to PCOS induced group (P>0.05). However, red clover (750 mg/kg) significantly increased HDL and decreased LDL levels when compared to PCOS induced group (P<0.05). Treatment with red clover reduced ovarian weight, volumes of ovarian, medulla, cortex and number of cysts and increased number of oocytes compared to PCOS group. Both red clover and clomiphene citrate could treat the letrozole induced PCOS in rats; however, red clover indicated antioxidant activities more than clomiphene citrate. Red clover may be used for discovering anti-PCOS drugs with lower side effects.
... The study [34] aimed to research the possible interaction of the flavonoid eriodictyol with the TRPV1 receptor and work out its putative antinociceptive and hyperthermic effects. Eriodictyol was ready to displace [3H]-resiniferatoxin binding and inhibit calcium influx mediated by capsaicin, predicting that eriodictyol acts as a TRPV1 antagonist. ...
Book
The highly reactive hydroxyl radical can cause cellular damage to DNA, lipids, proteins, and bone cells. These radicals are robust reactive oxygen species, and there is no specific enzyme to defend against them in the human body. Therefore, it is indispensable to discover some compounds with adequate scavenging capacity on these. This capacity of the extracts and polyherbal formulation made from Hibiscus and Amla was analyzed in this work. A method of extraction of solvents from Hibiscus and Amla is proposed in this work to yield 1023 mg and 956mg from 20g of Amla and Hibiscus. The samples showed in different chemical tests revealed active principles or phytoconstituents, viz., alkaloids, terpenoids, tannins, flavonoids, and steroids. Both the samples did not show any hemolytic activity. It showed only 26% hemolytic activity in the Hibiscus sample and 25% hemolytic activity in the Amla sample. The obtained data has negligible hemolysis activity. Amla has shown significant results with a lower IC50 value of 3.97µg/ml than Hibiscus in the anti-lipid peroxidation test. EC50 for Hibiscus was 213.70, while Amla showed a lower proliferation of 30% in cytotoxicity tests. The ALP activity release at the UMR-106 cell was 29.29 U/L at a 320µg/ml concentration for Amla and 20.85 U/L at 320µg/ml Hibiscus. Decreased release of ALP activity is noted only at higher concentrations for both Amla and Hibiscus. An increase in concentration has increased the calcium deposits in UMR-106 cells for both Amla and Hibiscus. Calcium deposits are a sign of successful invitro bone formation.
... Herbal medicinal extracts have received considerable attention within recent years, especially those containing phytoestrogens due to their beneficial effects in treating PCOS alongside having no major side effects (Abasian et al., 2018;Beck et al., 2005;Khani et al., 2011). Phytoestrogens are moderately estrogenic and could play a role in the prevention of other estrogen-related disorders, including cardiovascular diseases, osteoporosis and hormone-related cancers (Sunita and Pattanayak, 2011). ...
Article
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Ethnopharmacological relevance Chickpea was used in both greek and indian traditional medicine for hormonal related conditions as menstrual induction, acceleration of parturation, treatment of retained placenta and stimulation of lactation. Chickpea (Cicer arietinum) sprout isoflavone isolates exhibited reasonable estrogenic activities. Isoflavones, a subtype of phytoestrogens, are plant derivatives with moderate estrogenic activity that tend to have protective effects on hormonal and metabolic abnormalities of women with polycystic ovary syndrome (PCOS). Aim of the study In this study, we investigated the effect of UPLC/ESI-MS characterized Cicer arietinum L. seeds ethanol extract (CSE) on ovarian hormones, oxidative response and ovarian histological changes on induced PCOS rat model. Materials and methods Thirty-five rats were divided into five groups including negative control, PCOS, and treatment groups. PCOS was induced using letrozole (1 mg/kg) daily orally for 21 days. Each treatment group was treated with one of the following for 28 days after induction of PCOS: clomiphene citrate (1 mg/kg), and CSE at 250 and 500 mg/kg. Ovaries and uteri were excised, weighed and their sections were used for quantitative real-time reverse transcriptase polymerase chain reaction, antioxidant assays and histomorphometric study of the ovaries. The antioxidant assays, histopathological examination, hormonal and metabolic profiles, and Cyp11a1(steroidogenic enzyme) mRNA expression were measured. Results In all treatment groups, ovarian weight was significantly decreased despite having no significant effect on uterine weight. Histomorphometric study in the treatment groups revealed a significant decrease in the number and diameter of cystic follicles, a significant increase in granulosa cell thickness while, thickness of theca cells was significantly decreased when compared to PCOS. Hormone levels, metabolic profile and antioxidant status were improved in the treatment groups. Moreover, Cyp11a1 mRNA expression was significantly downregulated in the treatment groups compared to PCOS. Conclusions In the current study, CSE enhanced the reproductive and metabolic disorders which were associated with PCOS induction. For the first time, we have highlighted the effect of CSE in treating PCOS and its associated manifestations.
... However, some atrophic changes were still visible. The reduction in uterotrophic effects caused by Tualang honey in ovariectomised rats was important given that the risk of endometrial cancer is associated with oestrogen replacement therapy [60,61]. Therefore, honey can be recommended as an alternative therapy without significant side effects for HRT. ...
Article
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Nowadays, most people who lead healthy lifestyles tend to use natural products as supplements, complementary medicine or alternative treatments. Honey is God’s precious gift to mankind. Honey has been highly appreciated and extensively used since ancient history due to its high nutritional and therapeutic values. It is also known to enhance fertility. In the last few decades, the important role of honey in modern medicine has been acknowledged due to the large body of convincing evidence derived from extensive laboratory studies and clinical investigations. Honey has a highly complex chemical and biological composition that consists of various essential bioactive compounds, enzymes, amino and organic acids, acid phosphorylase, phytochemicals, carotenoid-like substances, vitamins and minerals. Reproductive health and fertility rates have declined in the last 30 years. Therefore, this review aimed to highlight the protective role of honey as a potential therapeutic in maintaining reproductive health. The main role of honey is to enhance fertility and treat infertility problems by acting as an alternative to hormone replacement therapy for protecting the vagina and uterus from atrophy, protecting against the toxic effects of xeno-oestrogenic agents on female reproductive functions and helping in the treatment of gynaecological disorders, such as vulvovaginal candidiasis infection, that affect women’s lives.
... Native Americans traditionally valued red clover for the treatment of external skin problems and lung, nervous and reproductive system ailments. The isoflavone constituents in red clover have estrogenic properties.They are thought to have positive effects on menopausal disorders such as osteoporosis, cardiac risk factors or breast cancer (Fugh-Berman and Kronenberg, 2001; Beck et al., 2005). It is also used as expectorant, analgesic, antiseptic ,tonic (Sabudak et al., 2009), sore throat, fever, pneumonia, and meningitis; skin problems and lung illnesses disorders of reproductive system (Kołodziejczyk-Czepas 2012). ...
... max) having several health promoting effects like cardiovascular disease, prevention of cancer, and reduction in osteoporosis. These are recommended to relieve for menopausal women from menopause symptoms instead of hormonal replacement therapy (HRT) (Beck et al., 2005). The HRT of isoflavones is due to the estrogenic effects of the metabolites. ...
Chapter
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Legumes are important source of nutrients in human diet, particularly from low-income group of developing nations. They are rich in proteins and good source of polyphenolic compounds which result in several positive effects on health. However, their nutritional value is adversely impacted by the presence of many antinutritional factors, such as amylase inhibitors, chlorogenic acid, cyanogenic glycosides, goitrogens, gossypol, isoflavones, lectins (phytohemagglutinins), oligosaccharides, oxalates, phytic acid (phytates), protease inhibitors (trypsin inhibitors), saponins, tannins, etc. These antinutritional substances can be removed or minimized to an accept- able level by simple and inexpensive processing techniques like boiling, dehulling, fermentation, extrusion, soaking, and pressure cooking which are discussed in detail. The present chapter will also highlight the significance of antinutritional substances in human health. Recent researches carried out to eliminate these antinutrients using technological processing are also emphasized in the study.
... These isoflavones possess estrogenic [5] and antiproliferative effects [6]. Various products from semi-purified isoflavones and their free forms have been studied for the possible preventive role of breast and prostate cancer, maintenance of bone health, improvement of cardiovascular health and benefits related to menopausal problems [5,[7][8][9][10]. ...
Article
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Isoflavones can be found in different chemical forms, but the health beneficial effects mainly appear in their free forms—aglycones. Their yield in red clover (Trifolium pratensis L.) extracts differs due to different extraction and hydrolysis methodologies. The main aim of this study was to obtain the highest yields of daidzein and genistein from red clover blossoms through the various extraction and hydrolysis methods and to increase their quantities using additional excipients. Extracts were obtained by ultrasound-assisted, heat-reflux and maceration methods combining them with acidic, alkaline, and thermal hydrolysis. Using ultrasound-assisted extraction with optimal conditions and heat-reflux method highest yields of isoflavones were obtained in UTE510 (393.23 ± 19.66 µg/g daidzein and 171.57 ± 8.58 µg/g genistein); UTE530 (415.07 ± 20.75 µg/g daidzein and 150.57 ± 7.53 µg/g genistein) and HNE5 (432.30 ± 21.61 µg/g daidzein and 154.50 ± 7.72 µg/g genistein) samples. These conditions were used with excipients: magnesium aluminometasilicate, croscarmellose sodium, sodium carboxymethyl starch and vinylpyrrolidone-vinyl acetate copolymer. This is the first study reporting the ability of the vinylpyrrolidone-vinyl acetate copolymer to promote solubilization and availability of active compounds from a herbal extract, resulting in enhanced isoflavones yield. The results of the present study showing increased solubility and availability provided by the vinylpyrrolidone-vinyl acetate copolymer suggest that this preparation could in principle also reduce variability due to limited water solubility of isoflavones.
... The study [34] aimed to research the possible interaction of the flavonoid eriodictyol with the TRPV1 receptor and work out its putative antinociceptive and hyperthermic effects. Eriodictyol was ready to displace [3H]-resiniferatoxin binding and inhibit calcium influx mediated by capsaicin, predicting that eriodictyol acts as a TRPV1 antagonist. ...
Book
Full-text available
The highly reactive hydroxyl radical can cause cellular damage to DNA, lipids, proteins, and bone cells. These radicals are robust reactive oxygen species, and there is no specific enzyme to defend against them in the human body. Therefore, it is indispensable to discover some compounds with adequate scavenging capacity on these. This capacity of the extracts and polyherbal formulation made from Hibiscus and Amla was analyzed in this work. A method of extraction of solvents from Hibiscus and Amla is proposed in this work to yield 1023 mg and 956mg from 20g of Amla and Hibiscus. The samples showed in different chemical tests revealed active principles or phytoconstituents, viz., alkaloids, terpenoids, tannins, flavonoids, and steroids. Both the samples did not show any hemolytic activity. It showed only 26% hemolytic activity in the Hibiscus sample and 25% hemolytic activity in the Amla sample. The obtained data has negligible hemolysis activity. Amla has shown significant results with a lower IC50 value of 3.97µg/ml than Hibiscus in the anti-lipid peroxidation test. EC50 for Hibiscus was 213.70, while Amla showed a lower proliferation of 30% in cytotoxicity tests. The ALP activity release at the UMR-106 cell was 29.29 U/L at a 320µg/ml concentration for Amla and 20.85 U/L at 320µg/ml Hibiscus. Decreased release of ALP activity is noted only at higher concentrations for both Amla and Hibiscus. An increase in concentration has increased the calcium deposits in UMR-106 cells for both Amla and Hibiscus. Calcium deposits are a sign of successful invitro bone formation.
... Isoflavone derivatives (isoflavones) are a class of flavonoids that belong to a large family of polyphenolic compounds from plants (such as soybeans, red clover, and kudzu roots) (7)(8)(9). Isoflavones are also called phytoestrogens because they contain an estrogen structure (particularly 17β-estradiol) (Fig. 1), and mimic the effects of estrogens by binding to estrogen receptors (10)(11)(12). Isoflavones are divided into a glycosidic form with sugar chains and an aglycone form without sugar chains in their molecular structures (8). ...
Article
The present study investigated the effects of isoflavone derivatives (daidzein, genistein and glycitein) on the production of inflammatory cytokines (IL-6 and IL-8) by IL-1β-stimulated synovial cells. Synovial MH7A cells were stimulated with IL-1β in the absence or presence of isoflavone derivatives, and IL-6 and IL-8 production was measured by ELISA. The results of the present study indicated that daidzein significantly inhibited the production of IL-6, but not IL-8. Conversely, neither genistein nor glycitein exerted any inhibitory effects on the production of IL-6 or IL-8 by IL-1β-stimulated synovial cells. To elucidate the molecular mechanisms underlying the daidzein-mediated inhibition of IL-6 production, the present study examined the effects of daidzein on the phosphorylation (activation) of NF-κB p65, ERK1/2 and p38 MAPK. Daidzein significantly inhibited the phosphorylation of NF-κB p65 and ERK1/2, but not p38 MAPK in IL-1β-stimulated MH7A cells. The present study revealed that among the isoflavone derivatives examined (daidzein, genistein and glycitein), daidzein inhibited the production of IL-6, but not IL-8, by IL-1β-stimulated synovial MH7A cells via the suppression of NF-κB p65 and ERK1/2 activation. Collectively, these results suggested that daidzein may have potential as a therapeutic agent for the treatment of arthritic disorders through its anti-inflammatory effects via the inhibition of IL-6 production.
... Gut microflora metabolizes phytoestrogen daidzein, found in T. pratense extract [19], into equol which has high affinity for estrogen receptor (ER) β [20]. Thus, the interaction between plant derived phytoestrogens with gut microbiota could yield positive estrogenic effects mainly in tissues expressing ER β which mediates cholesterol metabolism [21,22]. Prolonged consumption of herbal extract has been shown to regulate the composition of gut microbiota [23]. ...
Article
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Background Trifolium pratense (red clover) ethanolic extract (TPEE) has been used as a popular over-the-counter remedy for the management of menopausal symptoms. Prolonged consumption of herbal extract has been shown to regulate the composition of gut microbiota. This study was designed to elucidate the influence of TPEE on the gut microbiota composition in the ovariectomized (OVX) rats. Methods OVX rats were treated with TPEE at 125, 250, 500 mg/kg/day, or controls (pomegranate extract, 500 mg/kg/day; estradiol, 25 μg/kg/day) for 12 weeks. Gut microbiota analysis was conducted by extracting the microbial DNA from fecal samples and microbiome taxonomic profiling was carried out by using next-generation sequencing. The levels of serum biomarkers were analyzed using enzyme-linked immunosorbent assay (ELISA) kit. The prediction of functional biomarker of microbiota was performed using PICRUSt to investigate the potential pathways associated with gut health and serum lipid profile regulation. To study the correlation between gut microbiota composition and serum lipid levels, Spearman’s correlation coefficients were defined and analyzed. Additionally, gas chromatography–mass spectrometry analysis was conducted to uncover additional physiologically active ingredients. Results TPEE-treated OVX rats showed significant reduction in serum triglycerides (TG), total cholesterols (TCHOL), and LDL/VLDL levels but increase in HDL level. The alteration in the pathways involve in metabolism was the most common among the other KEGG categories. Particularly, TPEE also significantly reduced the relative abundance of sequences read associated with inflammatory bowel disease (IBD) and the peroxisome proliferator-activated receptor (PPAR) signalling pathway. TPEE intervention was seen to reduce the Firmicutes to Bacteroidetes (F/B) ratio in the OVX rats, denoting a reduction in microbial dysbiosis in the OVX rats. Correlation analysis at the phylum level revealed that Bacteriodetes and Proteobacteria were strongly correlated with serum TG, TCHOL and HDL levels. At the species level, Bifidobacterium pseudolongum group was seen to positively correlate with serum HDL level and negatively correlated with serum AST, ALT, LDL/VLDL, TCHOL, and TG levels. Conclusions TPEE treatment showed therapeutic benefits by improving the intestinal microbiota composition which strongly correlated with the serum lipid and cholesterol levels in the OVX rats.
... Legumes from the family Fabaceae are considered to be the main source of isoflavones, mainly Glycine max (soybean) containing genistein, glycitein, daidzein from Trifolium pratense (Red clover) as a source of biochanin A and formononetin [3]. These isoflavone compounds possess appreciable affinity to estrogen receptor (alpha), estrogen receptor (beta), androgen receptor and progesterone receptor, thus suggesting their high estrogenic activities [4]. Members of the family Fabaceae are rich in polyphenolic compound(s) that exhibit antibacterial activity [5]. ...
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Targeting quorum sensing controlled virulence factor in disease causing Pseudomonas aeruginosa has been an effective approach to cope with antibiotic resistance. Plant derivative compounds for example flavonoids and isoflavonoids have been identified as essential molecules of human diet and are known to possess various pharmacological properties. In this study, two flavonoids (apigenin and acacetin) and three isoflavonoids (genistein, daidzein and biochanin A) were investigated against N-acyl-homoserine lactone (AHL) intermediated virulence factor production in P. aeruginosa. The virulence factors that were focused in the current study are biofilm formation, synthesis of pyoverdin, pyocyanin, rhamnolipid as well as exo-polysaccharide. The RT-qPCR was also performed to analyse the virulence genes expression. Based on the experimental studies, it was observed that the flavone acacetin exhibited the strongest inhibition capacity towards all the bacterial virulence factors. Moreover, a significant down regulation in the expression of virulence genes upon treatment with the compounds was observed, suggesting that these compounds are effective targets for anti-quorum sensing approach.
... Currently, various parts of this medicinal plant are used in the management and treatment of many disorders, specifically digestive system disorders. Moreover, it is highly useful in the treatment of inflammation of bronchioles, chronic cough, diabetes mellitus, kidney stones as well as in nausea and vomiting [91]. ...
... Currently, various parts of this medicinal plant are used in the management and treatment of many disorders, specifically digestive system disorders. Moreover, it is highly useful in the treatment of inflammation of bronchioles, chronic cough, diabetes mellitus, kidney stones as well as in nausea and vomiting [87]. ...
... Currently, various parts of this medicinal plant are used in the management and treatment of many disorders, specifically digestive system disorders. Moreover, it is highly useful in the treatment of inflammation of bronchioles, chronic cough, diabetes mellitus, kidney stones as well as in nausea and vomiting [87]. ...
... With the discovery of the medicinal value of RC, many studies have shown that the chemical properties of isoflavones extracted from RC are similar to those of estrogen and can thus regulate estrogen levels in the human body. RC is one of the major sources of isoflavones (4,5). In recent years, RC has been used in ameliorating menopausal symptoms, such as hot flush, night sweats, urogenital atrophy, depression, and anxiety, in postmenopausal women (6). ...
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In red clover ( Trifolium pratense L.; RC) the main compound is isoflavones, which are selective estrogen receptor modulators for maintaining female health. Isoflavones exert antifatigue effects during exercise in high-temperature environments. This study aimed to investigate the effect of RC supplementation on gut microbiota composition to determine whether it improves intestinal barrier function and exercise performance. Female ICR mice were divided into four groups ( n = 8 per group) and orally administered RC once daily for 6 weeks at 0 (vehicle), 308 (RC-1X), 615 (RC-2X), and 1,538 (RC-5X) mg/kg. RC supplementation decreased the fat mass and increased exhaustive swimming time, grip strength, and muscle glycogen in female mice. In the RC supplementation group, serum levels of lactate, ammonia, and creatine kinase decreased after swimming. The estradiol and progesterone levels were higher in the RC group than in the vehicle group. Regarding gut microbiota composition, the RC-2X group may increase intestinal health related to the microorganisms Pseudobutyrivibrio and Parabacteroide . Thus, the use of RC supplements as nutraceuticals could have positive effects on athletes' gut and overall health.
... The fate of isoflavones of red clover derivatives such as Promensil is consistent in sheep and many other animal species with well-known metabolism, particularly formononetin and biochanin A (Beck, Rohr, and Jungbauer 2005;Fritz et al. 2013;Fugh-Berman 2007). An extremely low affinity for the estrogen receptor was observed when investigating the mechanism of action of formononetin and biochanin A (Murota, Nakamura, and Uehara 2018;Silva 2021). ...
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Isoflavones are secondary metabolites that represent the most abundant category of plant polyphenols. Dietary soy, kudzu, and red clover contain primarily genistein, daidzein, glycitein, puerarin, formononetin, and biochanin A. The structural similarity of these compounds to β-estradiol has demonstrated protection against age-related and hormone-dependent diseases in both genders. Demonstrative shreds of evidence confirmed the fundamental health benefits of the consumption of these isoflavones. These relevant activities are complex and largely driven by the source, active ingredients, dose, and administration period of the bioactive compounds. However, the preclinical and clinical studies of these compounds are greatly variable, controversial, and still with no consensus due to the non-standardized research protocols. In addition, absorption, distribution, metabolism, and excretion studies, and the safety profile of isoflavones have been far limited. This highlights a major gap in understanding the potentially critical role of these isoflavones as prospective replacement therapy. Our general review exclusively focuses attention on the crucial role of isoflavones derived from these plant materials and critically highlights their bioavailability, possible anticancer, antiaging potentials, and microbiome modulation. Despite their fundamental health benefits, plant isoflavones reveal prospective therapeutic effects that worth further standardized analysis.
Article
An efficient sample clean-up and preconcentration procedure for phytoestrogens analysis in urine has been developed. It was based on a combination of solid phase extraction with hollow-fiber supported liquid membrane and molecularly imprinted beads (MIPs-HF-SLM-SPE). The molecularly imprinted polymers (MIPs) were synthesized by precipitation polymerization technique with biochanin A (BCA) as a template, giving narrowly dispersed microspheres with a regular shape. As the functionalmonomer, (dimethylamino)ethyl methacrylate (DMA) turned out to be better than methacrylic acid (MAA) to get the best-imprinted effects. The MIPs used as sorbents in the MIPs-HF-SLM-SPE extraction process exhibited excellent binding selectivity for biochanin A, in comparison to nonimprinted polymers as well as its structural analogs (genistein and daidzein). Finally, the developed method was used to detect biochanin A in urine. Under optimal extraction conditions, the recovery of BCA in urine samples (using 4.5 mL sample spiked with 10 μg L-1) was over 41%, with a coefficient of variation (CV) < 6.6% (n=5). The detection limit (LOD) and quantification limit (LOQ) for BCA analysis in urine were 0.41 and 1.36 μg L-1, respectively.
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The theoretical study provides an overview of current data on red clover (Trifolium pratense L.), its cultivation and breeding, other practical uses and future prospects. Particular attention is paid to this taxon as a fodder plant of global importance in traditional and modern animal husbandry systems. Evidence is presented that red clover is a leading component, influencing the pasture management on all continents due to its high biomass production, protein content and overall nutritional value associated with the ability to fix atmospheric nitrogen. Some considerations cover important agrobiological and agrotechnological issues (e.g., plant interseeding) related to increasing adaptability, sustainability and yield of red clover and other crops coexisting in the same biosystems. Interesting biotechnological approaches have been pointed out in relation to the introduction to cultivation of new varieties, which are breeding hybrids of red clover and other wild species, with greater durability and yield-forming capacity. A separate part of the review is the assessment of the phytochemical profile of red clover with particular reference to biologically active specialized metabolites. Recent data from preclinical and human studies on existing and potential applications of this taxon and individual polyphenolic components in traditional and official medicine are also discussed.
Article
Trifolium pratense L. (Fam. – Fabaceae) whole plant’s powder is normally taken as tea to treat whooping and dry cough, and the ethanolic extract is used to check gouts and dermatitis complains. Besides the aforementioned Trifolium species, T. resupinatum, T. incarnatum, T. hybridum, T. pannonicum, T. purpureum, T. subterraneum, T. fragiferum, T. ambiguum, T. nigrescens, and T. medium were also grown in Mediterranean regions for feeding of sheep and cattle. The callus cultures of T. pratense, T. campestre, T. incarnatum, and T. rubens were established by inoculating the petioles on B5 and L5 medium. The Murashige and Skoog culture medium was supplemented with 1‐naphthaleneacetic acid and kinetin for enhancement of growth of callus.
Article
The importance of Trifolium pratense L. as a dietary supplement and its use in traditional medicine prompted the preparation of a thorough metabolite profile. This included the identification and quantitation of principal constituents as well as low abundant metabolites that constitute the residual complexity (RC) of T. pratense bioactives. The purity and RC of isoflavonoid fractions from standardized red clover extract (RCE) was determined using an off-line combination of countercurrent separation (CCS) and two orthogonal analytical methodologies: quantitative ¹H NMR spectroscopy with external calibration (EC-qHNMR) and LC-MS. A single-step hydrostatic CCS methodology (Centrifugal Partition Chromatography [CPC]) was developed that fractionated the isoflavonoids with a hexanes-ethyl acetate-methanol-water (HEMWat) 5.5/4.5/5/5, v/v solvent system (SS) into 75 fractions containing 3 flavonolignans, 2 isoflavonoid glycosides, as well as 17 isoflavonoids and related compounds. All metabolites were identified and quantified by qHNMR spectroscopy. The data led to the creation of a complete isoflavonoid profile to complement the biological evaluation. For example, fraction 69 afforded 90.5% w/w biochanin A (17), with 0.33% w/w of prunetin (16), and 0.76% w/w of maackiain (15) as residual components. Fraction 27 with 89.4% w/w formononetin (13) as the major component had, in addition, a residual complexity consisting of 3.37%, 0.73%, 0.68% w/w of pseudobaptigenin (11), kaempferol (10) and pratensein (8), respectively. Despite the relatively high resolving power of CPC, and not unexpectedly, the chromatographic fractions retained varying degrees of the original metabolomic diversity. Collectively, the extent of metabolomic diversity should be recognized and used to guide the development of isolation strategies, especially when generating samples for bioactivity evaluation. The simultaneous structural and quantitative characterization enabled by qNMR, supported by LC-MS measurements, enables the evaluation of a relatively large number of individual fractions and, thereby, advances both the chemical and biological evaluation of active principles in complex natural products.
Article
Phytoestrogen is considered a promising natural remedy for various postmenopausal symptoms. However, most studies have focused on common soybean, Glycine max. Here, for the first time, we evaluated the in vivo effect of a distinct species, small black bean (Rhynchosia volubilis), on ovariectomized mice and investigated its impact on the intestinal microbiota and metabolic status. Ovariectomized mice exhibited significant body weight gain, a typical postmenopausal symptom, and microbial changes, such as decreased α-diversity; changes in microbial composition, especially abundances of the families Desulfovibrionaceae and Mogibacteriaceae and genus Dorea; and decreased amino acid and short-chain fatty acid levels. Administration of small black bean extract restores body weight and gut microbial perturbation to levels similar to those observed under normal conditions. Our data suggest that small black bean extract is a potential candidate functional food for treating postmenopausal symptoms via remodeling of the intestinal microbiome.
Article
Ethnopharmacological relevance Echinops latifolius Tausch (ELT) is traditional Mongolian medicine in China, and often used to against osteoporosis, strengthen tendons and bones, clear bones heat. Aim of the study: To study efficacy of ELT on ovariectomized (OVX) rats and underly metabolic pathways related to trabecular micro-achitecture changing of OVX. Materials and methods Three-month-old female Wistar rats were randomly divided into 4 groups (n = 6) including normal group (without surgery), sham group (bilateral laparotomy), OVX group (bilateral ovariectomy), and ELT-treated groups (ELT-treated after bilateral ovariectomy). The effects of ELT on trabecular micro-achitecture and biochemical markers of OVX rat were investigated by dual-energy X-ray absorptiometry machine and Enzyme-linked immunosorbent assay (ELISA), respectively. Untargeted metabolomics strategy was applied to discover the potential biomarkers and related metabolic pathways involving the progression of OVX-induced osteoporosis. Results The trabecular micro-achitecture and biochemical markers of OVX rats were improved by ELT. We found 36 potential biomarkers and 21 related metabolic pathways were involved in progression of OVX-induced osteoporosis. Amino acids metabolism and glycerophospholipids metabolism were mainly intervened in ELT treatment on ovariectomized rats. The disordered amino acids and glycerophospholipids metabolism closely related to the imbalance between bone resorption and formation were reversed by administration of ELT, indicating that the influences of ELT on OVX rats’ trabecular micro-achitecture may possible be associated with intervening amino acids and glycerophospholipids metabolism. Conclusions This approach may provide the metabolomic perspective to link metabolic alterations and anti-osteoporosis action of ELT, to further explain how ELT works in postmenopausal patients with bone loss.
Chapter
The “antiquality” constituents frequently define management and utilization of forages. This chapter discusses significant organic components of forages worldwide that impact selection of the plants for forage management systems and in animal utilization and animal production. Included in the discussion are the tannins and phytoestrogens in many legumes; cyanogenic compounds in white clover, sorghum, and related grasses; alkaloids in a host of forages that affect intake, digestion, and animal health; and two nonphysiologic amino acids, mimosine and 5‐methyl‐L‐cysteine sulfoxide. The discussion addresses these antiquality components from the standpoint of their chemistry, genetics, and impact on the environment, forage management decisions, and animal responses. Tannins and polyphenols occur in leaves, stems, roots,and flowers of many forages. They are especially prevalent in forage legumes.
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Women’s life stages are based on their reproductive cycle. This cycle begins with menstruation and ends with menopause. Aging is a natural phenomenon that affects all humans, and it is associated with a decrease in the overall function of the organism. In women, aging is related with and starts with menopause. Also, during menopause and postmenopausal period, the risk of various age-related diseases and complaints is higher. For this reason, researchers were pushed to find effective remedies that could promote healthy aging and extended lifespan. Apitherapy is a type of alternative medicine that uses natural products from honeybees, such as honey, propolis, royal jelly, etc. Royal jelly is a natural yellowish-white substance, secreted by both hypopharyngeal and mandibular glands of nurse bees, usually used to feed the queen bees and young worker larvae. Over the centuries, this natural product was considered a gold mine for traditional and natural medicine, due to its miraculous effects. Royal jelly has been used for a long time in commercial medical products. It has been demonstrated to possess a wide range of functional properties, such as: antibacterial, anti-inflammatory, vasodilatative, hypotensive, anticancer, estrogen-like, antihypercholesterolemic, and antioxidant activities. This product is usually used to supplement various diseases such as cardiovascular disease, Alzheimer’s disease, sexual dysfunctions, diabetes or cancer. The main objective of this study is to highlight the effectiveness of royal jelly supplementation in relieving menopause symptoms and aging-related diseases. We also aimed to review the most recent research advances regarding the composition of royal jelly for a better understanding of the effects on human health promotion.
Article
The present study was undertaken to investigate the safety of kaempferol (KEM) and biochanin-A (BCA) following subacute exposure in mice. KEM and BCA were administered in three different doses by oral administration for 28 days. Evaluation of general toxicity parameters by examining the clinical signs, body weight, organ weights, haematological, biochemical, oxidative stress parameters, and histopathology was done. Administration of KEM and BCA for 28 days did not show any clinical signs of toxicity, nor any treatment-related changes in body weight and organ weights in comparison to control. The haematological parameters such as red blood cell, white blood cell, platelets count, haemoglobin (Hb) level, haematocrit, mean corpuscular haemoglobin concentration, red cell distribution width, and platelet distribution width did not show any change in the treated groups and control. Furthermore, different biochemical parameters like markers of the liver (alanine aminotransferase and aspartate aminotransferase), kidney (creatinine and urea), and heart (creatinine kinase-myocardial band and lactate dehydrogenase) injury along with other biochemical parameters showed nonsignificant differences between treated groups and control. Results of oxidative stress parameters in treated groups showed insignificant variations with control. The level of antioxidant enzymes such as superoxide dismutase and catalase were markedly increased in the treated groups; however, these were nonsignificant in comparison to control. In histopathology, evaluation of all vital organs, such as liver, kidney, heart, and lungs, did not show any morphological abnormalities and lesions in treated groups and control. The present study suggests that KEM and BCA have no adverse effects on the general physiology in mice.
Chapter
Small-seeded leguminous plants are a very abundant group of which only some are used in various ways. Particular attention has been paid to the use of small-seeded leguminous for the protection of marginal lands and soils. This chapter describes the importance of this species for consumption, forage, and medical purposes. The biological value of active substances found in small-seeded legumes is not fully understood. A number of species such as lucerne (Medicago sativa L), red clover (Trifolium pratense L.), sweet clover (Melilotus officinalis L.), goat’s-rue (Galega officinalis L.), and fenugreek (Trigonella foenum-graecum L.) provide the important raw material for the pharmaceutical industry. Small-seeded legumes can also be an important nutritional product and are used to overcome chronic malnutrition in countries with high food deficits. Lucerne leaf protein (Extract Fleur Lucerne) has a high content of not only lysine but also tryptophan and threonine, making lucerne a valuable additional ingredient as a food supplement in animal diets based mainly on cereals protein. Lucerne can be a dietary supplement of choice to tackle malnutrition and the lack of balanced diets for almost two billion people in the world.
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The absorption of isoflavones varies substantially among individuals. It is unknown whether isoflavone absorption differs between those originating from soy and those from red clover, which contain different mixtures of isoflavones. Because both soy and red clover are increasingly used in foods and supplements, these issues were studied in 14 subjects in a single-blind, randomized, placebo-controlled, crossover trial. Soybean isoflavone glycosides and red clover isoflavone aglycones were incorporated into a breakfast cereal and eaten daily for 2 wk each, separated by a 2-wk control or washout period. The 24-h excretions of isoflavones in urine were measured; similar to25% of each isoflavone was recovered in urine, suggesting that similar amounts were absorbed irrespective of their glycoside/aglycone nature or the differing compositions of their sources (daidzein and genistein in soy and formononetin and biochanin in red clover). Although interindividual variability was high, there was less intraindividual variability; the amounts excreted when subjects consumed the two sources of isoflavone were correlated (r = 0.69; P = 0.007).
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We examined protein kinase C (PKC) in the regulation of breast cancer cells by estrogen. Estrogen receptor (ER)- positive () MCF-7 and ER-negative () HCC38 cells were treated with 17-estradiol (E2 )o r E2-BSA, which cannot enter the cell. E2 and E2-BSA rapidly increased PKC- in both cells via phosphatidylinositol-dependent phospholipase C and G protein, but not phospholipase A2 or arachidonic acid. In MCF-7 cells, E2 and E2-BSA had comparable effects, maximal at 90 min. In HCC38 cells, PKC was maximal at 9 min, with E2-BSA more than E2. Tamoxifen blocked estrogen-dependent PKC in MCF-7 cells and reduced it in HCC38 cells. ER-antag- onist ICI 182780, ER-agonist diethylstilbestrol, and antibodies to ER and ER had no effect. E2 stimulated ( 3 H)thymidine incorporation in MCF-7 only; E2-BSA had no effect. Tamoxifen did not alter E2-dependent increases in MCF-7 cells, whereas ICI 182780 reduced DNA synthesis in control and E2-treated cultures. PKC activity was positively correlated with tumor severity in 133 breast cancer specimens and was greater in ER() tumors. Tamoxifen treatment reduced recurrence, and recurrent tumors had higher PKC activity. This indicates that E2 rapidly increases PKC activity via membrane pathways not involving ER or ER and suggests that tamoxifen works by reducing PKC activity through non-ER/ER-dependent mechanisms. (Endocrinology 144: 1812-1824, 2003)
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Due to the estrogenic properties of soy-derived isoflavones, many postmenopausal women are using these compounds as a natural alternative to hormone replacement therapy (HRT). How isoflavones impact breast cancer in postmenopausal women is important, because a majority of breast cancer cases occur in this age group. Chemical induction of mammary tumors in female rats has been used to determine that exposure of the mammary gland to soy isoflavones prior to tumor induction is protective against tumor formation. Here we investigate the effect of dietary genistein on mammary tumors that have already formed. The study was designed to determine the action of dietary genistein in a low endogenous estrogen environment as is observed in postmenopausal women. Animals were ovariectomized (OVX) after mammary tumor development and were then placed into one of three treatment groups: positive-control (OVX+ estradiol implant), genistein (OVX+ 750 p.p.m. genistein) and negative-control (OVX alone). Tumors were distinguished as malignant or benign by histopathological examination and were further characterized as either estrogen-dependent or estrogen-independent using immunohistochemistry to identify the presence of both estrogen receptor (ER) alpha and the progesterone receptor (PR). Genistein at 750 p.p.m. increased the weight of estrogen-dependent adenocarcinomas in ovariectomized rats compared with the negative-control animals. Genistein treatment also resulted in a higher percentage of proliferative cells in tumors and increased uterine weights when compared with negative-control animals. Collectively, these effects are probably due to the estrogenic activity of genistein. Plasma genistein concentrations in animals fed the isoflavone-containing diet were at physiological levels relevant to human exposure. Estradiol concentrations in ovariectomized animals not receiving an estradiol supplement were similar to those observed in postmenopausal women. The data suggest that in an endogenous estrogen environment similar to that of a postmenopausal woman, dietary genistein can stimulate the growth of a mammary carcinogen MNU-induced estrogen-dependent mammary tumors.
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Based on the epidemiological evidence for a relationship between consumption of certain foods and decreased cancer incidence in humans, an assay was developed to screen and fractionate plant extracts for chemopreventive potential. This assay measures effects on the metabolism of [3H]benzo(a)pyrene [B(a)P] in hamster embryo cell cultures. Screening of several plant extracts has generated a number of activity leads. The 95% ethyl alcohol extract of one of these actives, Trifolium pratense L. Leguminosae, red clover, significantly inhibited the metabolism of B(a)P and decreased the level of binding of B(a)P to DNA by 30 to 40%. Using activity-directed fractionation by solvent partitioning and then silica gel chromatography, a major active compound was isolated and identified as the isoflavone, biochanin A. The pure compound decreased the metabolism of B(a)P by 54% in comparison to control cultures and decreased B(a)P-DNA binding by 37 to 50% at a dose of 25 micrograms/ml. These studies demonstrate that the hydrocarbon metabolism assay can detect and guide the fractionation of potential anticarcinogens from plants. The ability of the isoflavone biochanin A to inhibit carcinogen activation in cells in culture suggests that in vivo studies of this compound as a potential chemopreventive agent are warranted.
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In laboratory animals, the consumption of soy protein, rather than animal protein, decreases serum cholesterol concentrations, but studies in humans have been inconclusive. In this meta-analysis of 38 controlled clinical trials, we examined the relation between soy protein consumption and serum lipid concentrations in humans. We used a random-effects model to quantify the average effects of soy protein intake on serum lipids in the studies we examined and used hierarchical mixed-effects regression models to predict variation as a function of the characteristics of the studies. In most of the studies, the intake of energy, fat, saturated fat, and cholesterol was similar when the subjects ingested control and soy-containing diets; soy protein intake averaged 47 g per day. Ingestion of soy protein was associated with the following net changes in serum lipid concentrations from the concentrations reached with the control diet: total cholesterol, a decrease of 23.2 mg per deciliter (0.60 mmol per liter); 95 percent confidence interval, 13.5 to 32.9 mg per deciliter [0.35 to 0.85 mmol per liter]), or 9.3 percent; low-density lipoprotein (LDL) cholesterol, a decrease of 21.7 mg per deciliter (0.56 mmol per liter); 95 percent confidence interval, 11.2 to 31.7 mg per deciliter [0.30 to 0.82 mmol per liter]), or 12.9 percent; and triglycerides, a decrease of 13.3 mg per deciliter (0.15 mmol per liter; 95 percent confidence interval, 0.3 to 25.7 mg per deciliter [0.003 to 0.29 mmol per liter]), or 10.5 percent. The changes in serum cholesterol and LDL cholesterol concentrations were directly related to the initial serum cholesterol concentration (P < 0.001). The ingestion of soy protein was associated with a nonsignificant 2.4 percent increase in serum concentrations of high-density lipoprotein (HDL) cholesterol. In this meta-analysis we found that the consumption of soy protein rather than animal protein significantly decreased serum concentrations of total cholesterol, LDL cholesterol, and triglycerides without significantly affecting serum HDL cholesterol concentrations.
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Classically, 17beta-estradiol (E2) is thought to control homeostatic functions such as reproduction, stress responses, feeding, sleep cycles, temperature regulation, and motivated behaviors through transcriptional events. Although it is increasingly evident that E2 can also rapidly activate kinase pathways to have multiple downstream actions in CNS neurons, the receptor(s) and the signal transduction pathways involved have not been identified. We discovered that E2 can alter mu-opioid and GABA neurotransmission rapidly through nontranscriptional events in hypothalamic GABA, proopiomelanocortin (POMC), and dopamine neurons. Therefore, we examined the effects of E2 in these neurons using whole-cell recording techniques in ovariectomized female guinea pigs. E2 reduced rapidly the potency of the GABAB receptor agonist baclofen to activate G-protein-coupled, inwardly rectifying K+ channels in hypothalamic neurons. These effects were mimicked by the membrane impermeant E2-BSA and selective estrogen receptor modulators, including a new diphenylacrylamide compound, STX, that does not bind to intracellular estrogen receptors alpha or beta, suggesting that E2 acts through a unique membrane receptor. We characterized the coupling of this estrogen receptor to a Galpha(q)-mediated activation of phospholipase C, leading to the upregulation of protein kinase Cdelta and protein kinase A activity in these neurons. Moreover, using single-cell reverse transcription-PCR, we identified the critical transcripts, PKCdelta and its downstream target adenylyl cyclase VII, for rapid, novel signaling of E2 in GABA, POMC, and dopamine neurons. Therefore, this unique Gq-coupled estrogen receptor may be involved in rapid signaling in hypothalamic neurons that are critical for normal homeostatic functions.
Article
Substantial evidence indicates that diets high in plant-based foods may explain the epidemiologic variance of many hormone-dependent diseases that are a major cause of mortality and morbidity in Western populations. There is now an increased awareness that plants contain many phytoprotectants. Lignans and isoflavones represent two of the main classes of phytoestrogens of current interest in clinical nutrition. Although ubiquitous in their occurrence in the plant kingdom, these bioactive nonnutrients are found in particularly high concentrations in flaxseeds and soybeans and have been found to have a wide range of hormonal and nonhormonal activities that serve to provide plausible mechanisms for the potential health benefits of diets rich in phytoestrogens. Data from animal and in vitro studies provide convincing evidence for the potential of phytoestrogens in influencing hormone-dependent states; although the clinical application of diets rich in these estrogen mimics is in its infancy, data from preliminary studies suggest beneficial effects of importance to health. This review focuses on the more recent studies pertinent to this field and includes, where appropriate, the landmark and historical literature that has led to the exponential increase in interest in phytoestrogens from a clinical nutrition perspective.
Article
Background: Isoflavone phytoestrogen therapy has been proposed as a natural alternative to hormone replacement therapy (HRT). HRT has a beneficial effect on bone, but few trials in humans have investigated the effects of isoflavones on bone. Objective: The objective of the study was to determine the effect on bone density of a red clover-derived isoflavone supplement that provided a daily dose of 26 mg biochanin A, 16 mg formononetin, 1 mg genistein, and 0.5 mg daidzein for 1 y. Effects on biochemical markers of bone turnover and body composition were also studied. Design: Women aged 49-65 y (n = 205) were enrolled in a double-blind, randomized, placebo-controlled trial; 177 completed the trial. Bone density, body composition, bone turnover markers, and diet were measured at baseline and after 12 mo. Results: Loss of lumbar spine bone mineral content and bone mineral density was significantly (P = 0.04 and P = 0.03, respectively) lower in the women taking the isoflavone supplement than in those taking the placebo. There were no significant treatment effects on hip bone mineral content or bone mineral density, markers of bone resorption, or body composition, but bone formation markers were significantly increased (P = 0.04 and P = 0.01 for bone-specific alkaline phosphatase and N-propeptide of collagen type I, respectively) in the intervention group compared with placebo in postmenopausal women. Interactions between treatment group and menopausal status with respect to changes in other outcomes were not significant. Conclusion: These data suggest that, through attenuation of bone loss, isoflavones have a potentially protective effect on the lumbar spine in women.
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Background. Prostate cancer is the most prevalent nonskin cancer among men in the United States and is the second leading cause of cancer deaths in men, The cause of prostate cancer remains obscure. Recently it was hypothesized that low levels of vitamin D, a hormone with potent antitumor properties, may increase the risk for clinical prostate cancer.
Book
Reviewers. Preface. 1. Historical Perspective. 2. Biosystematics and Interspecific Hybridization. 3. Reproductive Biology, Genetics, and Evolution. 4. Morphology and Physiology. 5. Management, Utilization, Quality, and Antiquality. 6. Fungal Diseases. 7. Virus Diseases. 8. Nematode Diseases. 9. Insect Pests. 10. Persistence. 11. Rhizobium Relationships. 12. Breeding Methodology. 13. Tetraploid Red Clover. 14. Tissue Culture. 15. Seed Production. 16. Germplasm Acquisition, Maintenance and Evaluation. 17. Future Trends. About the Authors.
Article
Historically, research into vascular effects of hormone replacement therapy (HRT) poignantly highlights the difficulties in extrapolating from medical research into clinical practice. Original observational trials encouraged great enthusiasm that HRT was protective against coronary heart disease (CHID) in post-menopausal women. This was supported by a plethora of beneficial mechanistic effects of HRT on the vascular system. In contrast, recent controlled trials have shown that a specific combined oral HRT actually causes a small increased risk of cardiovascular and cerebrovascular events, compared to placebo. The absolute risks were small, yet were accompanied by increased venous thrombosis and breast cancer. Although many controversies still remain regarding the vascular effects of specific types of HRT, currently, in my opinion, all HRT should be considered as causing a small increased risk of vascular events, until proven otherwise. However, the apparent lack of HRT effectiveness in the prevention of vascular disease should not deter enthusiasm for the use of established preventive therapies, especially lifestyle measures.
Article
CONTEXT: Despite decades of accumulated observational evidence, the balance of risks and benefits for hormone use in healthy postmenopausal women remains uncertain. OBJECTIVE: To assess the major health benefits and risks of the most commonly used combined hormone preparation in the United States. DESIGN: Estrogen plus progestin component of the Women's Health Initiative, a randomized controlled primary prevention trial (planned duration, 8.5 years) in which 16608 postmenopausal women aged 50-79 years with an intact uterus at baseline were recruited by 40 US clinical centers in 1993-1998. INTERVENTIONS: Participants received conjugated equine estrogens, 0.625 mg/d, plus medroxyprogesterone acetate, 2.5 mg/d, in 1 tablet (n = 8506) or placebo (n = 8102). MAIN OUTCOMES MEASURES: The primary outcome was coronary heart disease (CHD) (nonfatal myocardial infarction and CHD death), with invasive breast cancer as the primary adverse outcome. A global index summarizing the balance of risks and benefits included the 2 primary outcomes plus stroke, pulmonary embolism (PE), endometrial cancer, colorectal cancer, hip fracture, and death due to other causes. RESULTS: On May 31, 2002, after a mean of 5.2 years of follow-up, the data and safety monitoring board recommended stopping the trial of estrogen plus progestin vs placebo because the test statistic for invasive breast cancer exceeded the stopping boundary for this adverse effect and the global index statistic supported risks exceeding benefits. This report includes data on the major clinical outcomes through April 30, 2002. Estimated hazard ratios (HRs) (nominal 95% confidence intervals [CIs]) were as follows: CHD, 1.29 (1.02-1.63) with 286 cases; breast cancer, 1.26 (1.00-1.59) with 290 cases; stroke, 1.41 (1.07-1.85) with 212 cases; PE, 2.13 (1.39-3.25) with 101 cases; colorectal cancer, 0.63 (0.43-0.92) with 112 cases; endometrial cancer, 0.83 (0.47-1.47) with 47 cases; hip fracture, 0.66 (0.45-0.98) with 106 cases; and death due to other causes, 0.92 (0.74-1.14) with 331 cases. Corresponding HRs (nominal 95% CIs) for composite outcomes were 1.22 (1.09-1.36) for total cardiovascular disease (arterial and venous disease), 1.03 (0.90-1.17) for total cancer, 0.76 (0.69-0.85) for combined fractures, 0.98 (0.82-1.18) for total mortality, and 1.15 (1.03-1.28) for the global index. Absolute excess risks per 10 000 person-years attributable to estrogen plus progestin were 7 more CHD events, 8 more strokes, 8 more PEs, and 8 more invasive breast cancers, while absolute risk reductions per 10 000 person-years were 6 fewer colorectal cancers and 5 fewer hip fractures. The absolute excess risk of events included in the global index was 19 per 10 000 person-years. CONCLUSIONS: Overall health risks exceeded benefits from use of combined estrogen plus progestin for an average 5.2-year follow-up among healthy postmenopausal US women. All-cause mortality was not affected during the trial. The risk-benefit profile found in this trial is not consistent with the requirements for a viable intervention for primary prevention of chronic diseases, and the results indicate that this regimen should not be initiated or continued for primary prevention of CHD.
Article
Many women will at some stage consider taking hormone replacement therapy, but uncertainty about the risks and benefits makes this decision difficult
Article
Epidemiological evidence suggests that isoflavone phytoestrogens may reduce the risk of cancer, osteoporosis, and heart disease, effects at least partially mediated by estrogen receptors α and β (ERα and ERβ). Because isoflavone dietary supplements are becoming increasingly popular and are frequently advertised as natural alternatives to estrogen replacement therapy, we have examined the effects of one of these supplements on estrogen-dependent behavior and ERα- and ERβ-dependent gene expression in the brain. In the adult female rat brain, 17β-estradiol treatment decreased ERβ messenger RNA signal in the paraventricular nucleus by 41%, but supplement treatment resulted in a 27% increase. The regulation of ERβ in the paraventricular nucleus is probably via an ERβ-dependent mechanism. Similarly, in the ventromedial nucleus of the hypothalamus, supplement treatment diminished the estrogen-dependent up-regulation of oxytocin receptor by 10.5%. The regulation of oxytocin receptor expression in the ventromedial nucleus of the hypothalamus is via an ERα-dependent mechanism. Supplement treatment also resulted in a significant decrease in receptive behavior in estrogen- and progesterone-primed females. The observed disruption of sexual receptivity by the isoflavone supplement is probably due to antiestrogenic effects observed in the brain. These results suggest that isoflavone phytoestrogens are antiestrogenic on both ERα- and ΕRβ-dependent gene expression in the brain and estrogen-dependent behavior.
Article
The mortality and incidence of breast cancer are high in Western industrialized countries and relatively low in developing countries in Asia and other parts of the world. In Japan the mortality of breast cancer has gradually been increasing, but is still much lower compared with those of Western countries. Within Japan the mortality of breast cancer is higher in urban areas than in non-urban areas. The future estimation of cancer incidence in Japan predicts that breast cancer will become a leading cancer in the near future. From the present review of epidemiology of breast cancer it is recommended that excess intake of fat and calories, especially of animal fat, be avoided.
Article
Background: Ovarian cancer is the fourth most common cancer in women in the UK, with about 6700 developing the malignancy and 4600 dying from it every year. However, there is limited information about the risk of ovarian cancer associated with the use of hormone replacement therapy (HRT). Methods: 948,576 postmenopausal women from the UK Million Women Study who did not have previous cancer or bilateral oophorectomy were followed-up for an average of 5.3 years for incident ovarian cancer and 6.9 years for death. Information on HRT use was obtained at recruitment and updated where possible. Relative risks for ovarian cancer were calculated, stratified by age and hysterectomy status, and adjusted by area of residence, socioeconomic group, time since menopause, parity, body-mass index, alcohol consumption, and use of oral contraceptives. Findings: When they last reported HRT use, 287,143 women (30%) were current users and 186 751 (20%) were past users. During follow-up, 2273 incident ovarian cancers and 1591 deaths from the malignancy were recorded. Current users were significantly more likely to develop and die from ovarian cancer than never users (relative risk 1.20 [95% CI 1.09-1.32; p=0.0002] for incident disease and 1.23 [1.09-1.38; p=0.0006] for death). For current users of HRT, incidence of ovarian cancer increased with increasing duration of use, but did not differ significantly by type of preparation used, its constituents, or mode of administration. Risks associated with HRT varied significantly according to tumour histology (p<0.0001), and in women with epithelial tumours the relative risk for current versus never use of HRT was greater for serous than for mucinous, endometroid, or clear cell tumours (1.53 [1.31-1.79], 0.72 [0.52-1.00], 1.05 [0.77-1.43], or 0.77 [0.48-1.23], respectively). Past users of HRT were not at an increased risk of ovarian cancer (0.98 [0.88-1.11] and 0.97 [0.84-1.11], respectively, for incident and fatal disease). Over 5 years, the standardised incidence rates for ovarian cancer in current and never users of HRT were 2.6 (2.4-2.9) and 2.2 (2.1-2.3) per 1000, respectively-ie, one extra ovarian cancer in roughly 2500 users; death rates were 1.6 (1.4-1.8) and 1.3 (1.2-1.4) per 1000, respectively-ie, one extra ovarian cancer death in roughly 3300 users. Interpretation: Women who use HRT are at an increased risk of both incident and fatal ovarian cancer. Since 1991, use of HRT has resulted in some 1300 additional ovarian cancers and 1000 additional deaths from the malignancy in the UK.
Article
We investigated the estrogenic activities of isoflavone derivatives in competition binding assays with human estrogen receptor (hER) alpha or hER beta protein, and in a gene expression assay using a yeast system. Coumestrol binds as strongly as 17 beta -estradiol to both hERs. Biochanin A, 5-OMe-genistein, formononetin, and tectorigenin bind well to hER beta, but significant binding to hER alpha is only observed with 5-OMe-genistein, formononetin and tectorigenin. The binding of 7-OMe-genistein and irisolidone is poor to both receptors. Among the glucosides, sissotorin binds both receptors and the binding is stronger than genistin. Coumestrol induces transcription as strongly as genistein. Tectorigenin also induces transcription with both hERs. Though biochanin A, 5-OMe-genistein, 7-OMe-genistein, irisolidone and formononetin slightly induce transcription with hER P, they act as antagonists in the induction of transcription by 17 beta -estradiol. The results show that methylation or glucoidation of isoflavones generally inhibits their phytoestrogenic activities.
Article
A novel estrogen receptor, estrogen receptor β (ERβ), has recently been cloned from a rat prostate cDNA library. In bone, which is an important target tissue of estrogen, ERα has been reported to be present preferentially in osteoblasts, but the mechanism of action of estrogen in bone is still not known. In the present study, we examined expression of ERβ mRNA in bone. Expression of ERβ mRNA was evident in primary osteoblastic cells isolated from 1-day-old rat calvaria and rat osteosarcoma cells (ROS 17/2.8), and its level was higher than that of ERα mRNA. When osteoblastic cells were cultured for 28 days to induce differentiation into mature osteoblasts capable of forming bone nodules, ERβ mRNA was constantly and highly expressed during the entire culture period. In contrast, the level of ERα mRNA was very low at the beginning of culture and it gradually increased during the differentiation of osteoblastic cells. Various tissues including bone were isolated from 8-week-old rats of both sexes, and total RNA was extracted to compare the tissue distribution of expression levels of ERβ mRNA. In cancellous bone of the distal femoral metaphysis and lumbar vertebra, expression of ERβ mRNA was obvious, and its level was equivalent to those in the uterus and testis, but lower than those in the ovary and prostate. The level of ERβ mRNA in femoral cortical bone was lower than that in cancellous bone. There was no appreciable differences between female and male rats in the distribution and expression levels of ERβ mRNA in bone. These results indicate that ERβ mRNA is highly expressed in osteoblasts in rat bone, suggesting that there is a distinct mechanism of estrogen action mediated by ERβ in bone.
Article
Dietary factors are considered important environmental risk determinants for Western diseases. Studies have revealed beneficial or protective effects of the consumption of legumes with regard to hypercholesterolaemia and coronary heart disease, obesity, diabetes mellitus, and menopause. During the last decade attention has been focused on soy and soybean products. Several constituents have been isolated: isoflavones, phytosterols, protease inhibitors, inositol hexaphosphate, and saponins. Our interest concentrates on hormone-like bisphenolic phytoestrogens of dietary origin, the lignans and isoflavonoids. Their glycosides, converted by gut bacteria to mammalian derivatives with weak estrogenic and antioxidative activity, originate in leguminous seeds. We developed an isotope dilution gas chromatography-mass spectrometry method for quantitative determination of the isoflavones, formononetin, biochanin A, daidzein, genistein and coumestrol, and the lignans secoisolariciresinol and matairesinol, in food samples. We measured the four isoflavonoids and coumestrol, and, for the first time, the two lignans in 52 leguminous seeds and found high concentrations of isoflavonoids (0–1853.35 mg/kg; 0–7.3 mmol/kg dw) but lower amount of lignans (0–15.85 mg/kg; 0.05 mmol/kg dw). The highest plasma levels of their metabolites are found in individuals living in countries or regions with low cancer and cardiovascular disease incidence and these are probably sufficient to influence intracellular enzymes, protein synthesis, growth factor action, malignant cell proliferation, differentiation, and angiogenesis. Leguminous seeds, therefore, in respect to their abundant concentrations of phytoestrogens, are strong candidates for a role as natural cancer-protective food.
Article
Historically, research into vascular effects of hormone replacement therapy (HRT) poignantly highlights the difficulties in extrapolating from medical research into clinical practice. Original observational trials encouraged great enthusiasm that HRT was protective against coronary heart disease (CHD) in post-menopausal women. This was supported by a plethora of beneficial mechanistic effects of HRT on the vascular system. In contrast, recent controlled trials have shown that a specific combined oral HRT actually causes a small increased risk of cardiovascular and cerebrovascular events, compared to placebo. The absolute risks were small, yet were accompanied by increased venous thrombosis and breast cancer.Although many controversies still remain regarding the vascular effects of specific types of HRT, currently, in my opinion, all HRT should be considered as causing a small increased risk of vascular events, until proven otherwise. However, the apparent lack of HRT effectiveness in the prevention of vascular disease should not deter enthusiasm for the use of established preventive therapies, especially lifestyle measures.
Article
We have cloned a novel member of the nuclear receptor superfamily. The cDNA of clone 29 was isolated from a rat prostate cDNA library and it encodes a protein of 485 amino acid residues with a calculated molecular weight of 54.2 kDa. Clone 29 protein is unique in that it is highly homologous to the rat estrogen receptor (ER) protein, particularly in the DNA-binding domain (95%) and in the C-terminal ligand-binding domain (55%). Expression of clone 29 in rat tissues was investigated by in situ hybridization and prominent expression was found in prostate and ovary. In the prostate clone 29 is expressed in the epithelial cells of the secretory alveoli, whereas in the ovary the granulosa cells in primary, secondary, and mature follicles showed expression of clone 29. Saturation ligand-binding analysis of in vitro synthesized clone 29 protein revealed a single binding component for 17beta -estradiol (E2) with high affinity (Kd = 0.6 nM). In ligand-competition experiments the binding affinity decreased in the order E2 > diethylstilbestrol > estriol > estrone > 5alpha -androstane-3beta ,17beta -diol >> testosterone = progesterone = corticosterone = 5alpha -androstane-3alpha ,17beta -diol. In cotransfection experiments of Chinese hamster ovary cells with a clone 29 expression vector and an estrogen-regulated reporter gene, maximal stimulation (about 3-fold) of reporter gene activity was found during incubation with 10 nM of E2. Neither progesterone, testosterone, dexamethasone, thyroid hormone, all-trans-retinoic acid, nor 5alpha -androstane-3alpha ,17beta -diol could stimulate reporter gene activity, whereas estrone and 5alpha -androstane-3beta ,17beta -diol did. We conclude that clone 29 cDNA encodes a novel rat ER, which we suggest be named rat ERbeta to distinguish it from the previously cloned ER (ERalpha ) from rat uterus.
Article
The determination of oestrogenic isoflavones and commestrol in clover (Trifolium pratense L. and Trifolium repens L.)Thirty-two varieties of white clover and twenty varieties of red clover were examined for five isoflavones —biochanin A, formononetin, pratensein, genistein, daidzein— and the coumarin derivate coumestrol known as oestrogenic. A method for the extraction and the cleaning by column and thin-layer chromatography is described. The quantitative analysis is carried out spectrophotometrically. The determination of small amounts of formononetin and coumestrol in white clover is effected semi-quantitatively. The problems with the preparation of the sample, the extraction, the column and thin-layer chromatography and the elution are discussed. The chemical examination of the six oestrogenic substances in clover allows a better judgement whether the reproductive abnormalities for animals must be attributed to these substances or to other causes.ZusammenfassungFür die Bestimmung der fünf als östrogen bekannten Isoflavone —Biochanin A. Formononetin, Pratensein, Genistein, Daidzein— und des Cumarinderivatives Cumöstrol in 32 Weissklee- und 20 Rotkleesorten wird eine Extraktionsmethode, sowie ein säulen- und dünnschichtchromatographisches Reinigungsverfahrën beschrieben. Die quantitative Untersuchung wird spektrophotometrisch durchgeführt. Für die Bestimmung geringer Mengen Formononetin und Cumöstrol in Weissklee wird eine halbquantitative Methode angegeben. Die bei der Probenaufbereitung, Extraktion, Säulen- und Dünnschichtchromatographie sowie bei der Elution auftretenden Probleme werden diskutiert. Die chemische Untersuchung der sechs östrogenen Stoffe in Klee ermöglicht eine bessere Beurteilung, ob eventuelle Fruchtbarkeitsstörungen in der Viehhaltung auf diese Substanzen oder andere Ursachen zurückzuführen sind.
Article
Estrogens used in hormone replacement therapy regimens may increase the risk of developing breast cancer. Paradoxically, high consumption of plant-derived phytoestrogens, particularly soybean isoflavones, is associated with a low incidence of breast cancer. To explore the molecular basis for these potential different clinical outcomes, we investigated whether soybean isoflavones elicit distinct transcriptional actions from estrogens. Our results demonstrate that the estrogen 17β-estradiol effectively triggers the transcriptional activation and repression pathways with both estrogen receptors (ERs) ERα and ERβ. In contrast, soybean isoflavones (genistein, daidzein, and biochanin A) are ERβ-selective agonists of transcriptional repression and activation at physiological levels. The molecular mechanism for ERβ selectivity by isoflavones involves their capacity to create an activation function-2 surface of ERβ that has a greater affinity for coregulators than ERα. Phytoestrogens may act as natural selective estrogen receptor modulators that elicit distinct clinical effects from estrogens used for hormone replacement by selectively recruiting coregulatory proteins to ERβ that trigger transcriptional pathways.
Article
The concentrations of the isoflavones biochanin A, genistein, formononetin, pratensein and daidzein in leaves of four varieties of red clover and two of subterranean clover have been measured by a paper chromatographic-spectrophotometric method. Biochanin A and formononetin are the predominant isoflavones in red clovers whereas biochanin A and genistein are the most abundant in subterranean clovers. For both species the isoflavone levels differed between varieties and between sampling dates. Higher levels were found during flowering for red clovers and before flowering for subterranean clovers. The isoflavone contents have been calculated to have oestrogenic activity equivalent to 2–17 μg. of stilboestrol (% dry wt.) for the red clover samples and 22–48 μg. of stilboestrol (% dry wt.) for the subterranean clover samples. The potency of some of these samples is much higher than potencies previously reported in the literature.
Article
The effects of isoflavones (genistein and daidzein) on endometrial carcinogenesis in mice were investigated in two experiments. In the short-term experiment (2 weeks), single subcutaneous (s.c.) administration of genistein [1 mg/30 g body weight (b.w.)] significantly decreased the levels of estradiol-l7β (E2) (5 ppm in diet)-induced expression of c-jun, interleukin-lα (IL-lα) and tumor necrosis factor-a (TNF-a) mRNAs in the uteri of ovariectomized mice (P<0.005, P<0.05 and P<0.01, respectively). Daidzein significantly inhibited E2-induced expression of c-fos and IL-lα (P<0.01, P<0.01 respectively). In the long-term experiment (30 weeks), 140 female ICR mice were given N-methyl-N-nitrosourea-containing solution (1 mg/100 g b.w.) and normal saline (as controls) into their left and right uterine corpora, respectively. They were divided into six groups; group 1 was given E2 (in diet) alone. Group 2 was given E2 and genistein (1 mg/30 g b.w., s.c., every four weeks). Group 3 was exposed to E2 and daidzein (1 mg/30 g b.w., s.c., every four weeks). Groups 4 and 5 respectively received genistein and daidzein, and were kept on the basal diet. Group 6 was kept on the basal diet and served as a control. At the termination of the experiment, incidences of endometrial adenocarcinoma and atypical endometrial hyperplasia of the group given E2 and genistein or daidzein were significantly lower than of the group with E2 alone (P<0.01 and P<0.05, respectively). It is suggested that both genistein and daidzein have an inhibitory effect on estrogen-related endometrial carcinogenesis in mice, possibly by suppressing expression of estrogen-induced estrogen-related genes c-fos and c-jun, and internal cytokines IL-lα and TNF-α through a cytokine and estrogen receptor-mediated pathway.
Article
Background: Prostate cancer is the most prevalent nonskin cancer among men in the United States and is the second leading cause of cancer deaths in men. The cause of prostate cancer remains obscure. Recently it was hypothesized that low levels of vitamin D, a hormone with potent antitumor properties, may increase the risk for clinical prostate cancer. Methods: Because the major source of vitamin D is casual exposure to ultraviolet (UV) radiation, the authors examined the geographic distributions of UV radiation and prostate cancer mortality in 3073 counties of the contiguous United States using linear regression and trend surface analyses. Results: The geographic distributions of UV radiation and prostate cancer mortality are correlated inversely (P < 0.0001). Prostate cancer mortality exhibits a significant north-south trend, with lower rates in the South. These geographic patterns are not readily explicable by other known risk factors for prostate cancer. Conclusions: These data lend support to the hypothesis that UV radiation may protect against clinical prostate cancer. Viewed in conjunction with other recent data, including those demonstrating a differentiating effect of vitamin D on human prostate cancer cells, these findings suggest that vitamin D may have an important role in the natural history of prostate cancer.
Article
BACKGROUND The authors previously reported an increased risk of breast carcinoma with longer duration of hormone replacement therapy (HRT) use. It is unclear if different types of HRT confer different risks.METHODS In this study, a population-based cohort of 29,508 women were interviewed during 1990–1992 to determine whether there are any differences in breast carcinoma risk according to different types and duration of HRT use.RESULTSAt the end of the follow-up period in December 2001, the cohort constituted 298,649 person-years. Slightly more breast carcinoma cases were seen (n = 556) than expected (n = 508.37; standardized morbidity ratio =1.09, 95% confidence interval [CI] = 1.00–1.19). Approximately 3663 women had ever used HRT. In Cox regression models, time to breast carcinoma in relation to duration and type of HRT use was analyzed, adjusting for age at menarche, age at first full-term pregnancy, parity, age at menopause, family history of breast carcinoma, and age at interview. In women with a natural menopause, a significantly higher risk was observed for longer duration of combined continuous HRT use compared with never users (hazard ratio [HR] = 4.60, 95% CI = 2.39–8.84). Nonsignificant elevated risks also were observed for longer combined sequential (HR = 2.23, 95% CI = 0.90–5.56), gestagen only (HR = 3.74,9 5% CI = 0.94–14.97), and estriol use (HR = 1.89, 95% CI = 0.81–4.39). No increased risk was seen in women after 5 years of nonuse. When studying women who ever used only one type of HRT, even more elevated HRs for gestagen-containing preparations were seen. The highest risks were associated with the combined continuous and gestagen-only therapy in women with ≥ 48 months of use. Use of estradiol without progestins did not increase breast carcinoma risk significantly. The authors estimated the cumulative risk of breast carcinoma in a 50-year-old woman with gestagen-containing therapies for ≥ 48 months, with a follow-up of 10 years, to be 7% (95% CI = 5.4–11.4%) compared with 2% (95% CI = 1.6%–2.9%) for never-users of HRT.CONCLUSIONS Longer use of HRT containing progestins significantly elevates breast carcinoma risk whereas estradiol use does not. Continued use of progestins rendered the highest risks. The yearly risk of breast carcinoma for long-term users of progestins is of the magnitude of 50% the risk of a BRCA1 mutation carrier. Cancer 2003;97:1387–92. © 2003 American Cancer Society.DOI 10.1002/cncr.11205