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Ameliorative Potential of Methanolic Extract of Citrullus lanatus (Watermelon) Seeds on the Sperm Parameters, Testosterone Level and Testicular Cytoarchitecture of Male Albino Rats Induced with Lead-Acetate

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*Corresponding author: Email: nkwilly@gmail.com;
British Journal of Pharmaceutical Research
6(1): 35-43, 2015, Article no.BJPR.2015.047
ISSN: 2231-2919
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www.sciencedomain.org
Ameliorative Potential of Methanolic Extract of
Citrullus lanatus (Watermelon) Seeds on the Sperm
Parameters, Testosterone Level and Testicular
Cytoarchitecture of Male Albino Rats Induced with
Lead-Acetate
Onyeso Godspower
1
, Nkpaa Kpobari Williams
2*
and Nwaka Elochukwu
1
1
Department of Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences,
Madonna University, Elele, Rivers State, Nigeria.
2
Department of Biochemistry (Toxicology Unit), Faculty of Chemical Science, College of Natural and
Applied Science, University of Port Harcourt, P.M.B. 5323, Choba, Rivers State, Nigeria.
Authors’ contributions
This work was carried out in collaboration between all authors. Author OG designed the study, wrote
the protocol, carried out the statistical analysis and wrote the first draft of the manuscript. Author NKW
managed the literature searches and experimental analysis and author NE also managed the
experimental process. All authors wrote, read and approved the final manuscript.
Article Information
DOI:10.9734/BJPR/2015/15358
Editor(s):
(1)
Wenbin Zeng, School of Pharmaceutical Sciences, Central South University, Hunan, China.
Reviewers:
(1)
Armando Zarrelli, Department of Chemical Sciences, University Federico II, Complesso Universitario Monte S. Angelo, Italy.
(2)
Ahmed E. Abdel Moneim, Department of Zoology, Helwan University, Egypt.
Complete Peer review History:
http://www.sciencedomain.org/review-history.php?iid=982&id=14&aid=8104
Received 22
nd
November 2014
Accepted 31
st
December 2014
Published 10
th
February 2015
ABSTRACT
The study aimed to evaluate the ameliorative potential of methanolic extract of Citrullus lanatus (C.
lanatus; watermelon) seeds on lead-acetate induced testicular toxicity considering the sperm
parameters, testosterone level and testicular cytoarchitecture on adult male albino rats. The results
showed statistically significant (p<0.05) decrease in serum level of testosterone and a deleterious
effect on the sperm motility, count, morphology, viabilityand seminiferous tubular derangement on
lead-acetate treated rats when compared with the control group. The methanolic extract of C.
lanatus seed from the results showed a corrective effect as against the lead-acetate treated group
in relative to the control group. In conclusion it was discovered that methanolic extract of C. lanatus
Original Research Article
Godspower et al.; BJPR, 6(1): 35-43, 2015; Article no.BJPR.2015.047
36
seed has ameliorative potentials to correct the deleterious effect of lead-acetate on male
reproductive system.
Keywords: Citrullus lanatus seeds; lead-acetate; testicular toxicity; sperm parameters; testosterone
level; testicular cytoarchitecture.
1. INTRODUCTION
In recent years, more and more attention is being
paid to the regulation of spermatogenesis and to
the possibilities of influencing therapeutically this
somatic function in a positive or negative way.
However, human fecundity appears to be on the
decline [1], which cannot solely be attributable to
an increase in contraception. Rather, a body of
data suggests that poor semen quality and low
sperm count is markedly increasing and is likely
to be a contributing factor [1]. Although many
people still think of infertility as a problem that
affect only women, but in about 40% to 50% of
infertile couples, the man is the sole cause or a
contributing cause of the inability to conceive [2].
Exposure of heavy metals during pregnancy has
been associated with adverse effects on
development of gonads. These substances may
act as testicular toxicants and correspond to
different compounds, which are related to social
habits, life conditions, working hazards or use of
drugs and medicines [3,4]. Although, many
studies have reported the toxic and carcinogenic
effects of metals in human and animals, it is also
well known that these metals form a crucial part
in normal biological functioning of cells [3].
Lead represents a significant ecological and
public health concern due to its toxicity and its
ability to accumulate in living organisms. Earlier
studies have demonstrated that lead can pass
through the blood testis barrier, accumulate in
the testis and/or epididymis and seriously affect
the spermatogonia, primary spermatocytes,
spermatids or spermatozoa (germinal cells
different levels of differentiation) [5,6]. Several
studies assessed the genotoxic effect of lead
acetate (LA) by means of chromosomal
aberrations and micronucleus test. Regarding the
induction of chromosomal aberrations, LA
induced significant increase of aberrant cells and
numerical distortion in bone marrow cells of
Wistar rats [7,8]. Additionally, Aboul-Ela [9]
detected a significant increase of structural
chromosomal aberrations in bone marrow cells
and primary spermatocytes of albino mice
treated with LA. In addition, LA proved to be a
potent micronuclei inducer in vivo and in vitro test
systems: LA induced micronuclei in kidney cells
of Sprague–Dawley albino rats [10]; in human
melanoma cell [11]; in Chinese hamster V79
cells [12,13]; in Wistar rats’ leukocytes,
reticulocytes and erythrocytes [14,15]; in rats’
erythrocytes [16-18]; in peripheral blood
erythrocytes tissues, gill and fin epithelial cells of
Carassius auratus [19] and in bone marrow cells
of Algerian mice [20]. More so, the results of the
studies indicated that occupational exposure to
Pb has adverse effects on sperm parameters
(decreased sperm counts, lower and a lesser
motility and altered sperm morphology), studies
showed that exposure to inorganic lead greatly
impaired male reproductive functional activities
by decreasing sperm count or distorting sperm
motility and morphology [21].
The fruits of C. lanatus (watermelon) help in
Boosting Antioxidant Levels because it is
exceptionally rich in carotenoids such as
lycopene, lutein and β carotene [22]. A regular
watermelon juice consumption result in
significant increases in blood plasma
concentrations of lycopene and β carotene [23].
Also, lycopene is known to have over 40
potential health benefits and β carotene with
equally plentiful health benefits, which make this
finding very plausible. Moreover, watermelon-
induced is also known to increase plasma
antioxidant levels and may explain why
epidemiological studies of the Chinese found
greater watermelon intake to be associated with
a lower risk of cancer [24]. A study found that 6
weeks of treatment with a watermelon extract
containing 6 grams of L-citrulline and L-arginine
daily on middle-aged obese subjects with
prehypertension or stage 1 hypertension
experienced reduced ankle blood pressure and
altered carotid wave reflection, an indication of
improved arterial function of the individuals [25].
If watermelon can cure or ameliorate this
process, it would certainly provide a
breakthrough to many of the drugs used in the
market for primary prevention, such as the
cholesterol-lowering statin drug class, whose
side effects are numerous [26].
It is very important to note that all parts of the
watermelon have something to offer. For
Godspower et al.; BJPR, 6(1): 35-43, 2015; Article no.BJPR.2015.047
37
example, the seeds are excellent source of
protein. The good nutritional and functional
properties of watermelon seed meal proteins
suggest their potential use in food formulations
and diets [27]. C. lanatus possesses numerous
bioactivities from natural source which is of better
advantage than conventional therapies. This
study aimed to evaluate the ameliorative
potential of methanolic extract of C. lanatus
(watermelon) seeds on Lead-acetate induced
testicular toxicity considering the sperm
parameters, testosterone level and testicular
cytoarchitecture on adult male albino rats.
2. MATERIALS AND METHODS
2.1 Experimental Animals
A total number of 20 male Albino rats weighing
between 155 to 328 grams were used for the
study. The animals were bought from the animal
house of the Department of Pharmacology and
Toxicology, located in Niger Delta University
Wilberforce Island. It was kept in the animal
house of Department of Human Physiology
Madonna University, Elele Campus for 3 weeks
to acclimatize. The animals were kept under
normal room condition of temperature 25±2°C,
humidity of 50±5% and 12 hour light and dark
cycles. The animals were randomized into
experimental and control groups and housed in
sanitized wooden cages containing saw-dusts as
bedding. They were also fed with standard rat
chow pellet as diet and clean water ad-libitum
was supplied.
2.2 Seeds Collection and Preparation
Ripe watermelon pods were obtained from the
local market in Elele (Eke-Onuma Market),
Rivers State, Nigeria (May, 2013). The seeds
extracted from the pods, only healthy looking
seeds were collected. The collected seeds were
oven-dried at 35°C, to a constant weight. The
dried seeds were reduced into fine powder using
a Laboratory grinding hand mill. The powder was
weighed and kept away from light before
extraction.
2.3 Seeds Extraction and Concentration
Extraction was by maceration over a 72 hours
period. 500 g of the powdered seeds material
was extracted with 1.5 liters of Methanol in 3
successive extractions (500 ml every 24 hours).
The jar was tightly closed and thoroughly shaken
intermittently. After 72 hours, the different
portions were combined and filtered using filter
paper. The filtrate was collected in a glass jar.
The extract was concentrated using a Rotary
Evaporator. The concentrated methanolic extract
of Citrullus lanatus seeds were then transferred
into bottles covered with aluminium foil and
stored in a refrigerator at 4°C before use. This is
to prevent it from losing its potency.
2.4 Experimental Procedure
On commencement of the experiment, the
animals were divided into 4 groups of 5 animals
each. The first group served as the control group
while the last 3 groups served as the
experimental groups.
2.4.1 Group 1
This group was the control group fed normal rat
chow and water.
2.4.2 Group 2
This group was fed normal rat chow and water
and received 2.25 mg/kg of Lead-acetate + 100
mg/kg (Low dose) of methanolic extract of C.
lanatus seeds.
2.4.3 Group 3
This group was fed normal rat chow and water
and received 2.25 mg/kg of Lead-acetate + 200
mg/kg (High dose) of methanolic extract of C.
lanatus seeds.
2.4.4 Group 4
This group was fed normal rat chow and water
ad-libitum and they received 2.25 mg/kg Lead-
acetate only.
2.5 Analytical Procedure
After 30 days of treatment, the animals were
fasted for 24 hours prior to sacrifice. The animals
were anaesthetized using chloroform and then
sacrificed. Thus, blood was obtained via cardiac
puncture and put in labeled EDTA bottles for
testosterone assay. The animals were then
dissected; the testes were removed along with
the caudal epididymis. The caudal epididymis
was separated from the testes and lacerated to
collect the semen with a microscope glass slide
for analysis of sperm characteristic and the
Godspower et al.; BJPR, 6(1): 35-43, 2015; Article no.BJPR.2015.047
38
testes for testicular cytoarchitecture
(histopathology).
2.5.1 Estimation of serum testosterone
Quantitative measurement of serum total
testosterone was carried out adopting ELISA
technique using kits specific for rats purchased
from Department of Pharmacology and
Toxicology, Niger Delta University Wilberforce
Island, Nigeria according to the protocol provided
with kit.
2.5.2 Histological examination
The testes tissues were collected and
immediately fixed in 1 ml PBS (pH 7.4) and
embedded in paraffin. Sections (5-µm) were
prepared and then stained with hematoxylin and
eosin (H & E) stain for photomicroscopic
observations.
2.6 Statistics
The result were expressed as mean ± standard
error of mean (S.E.M). The statistical evaluation
of data was performed by using a one-way
ANOVA (analysis of variance). The data were
considered significant at p˂0.05.
3. RESULTS
As shown in Table 3.1, the level of active sperm
cells in Lead-acetate + HDE of C. lanatus seed
(65.00±2.88%) was significantly higher (p<0.05)
than the Lead-acetate only (11.66±7.26%). The
level of dead cells increased significantly
(p<0.05) in Lead-acetate only rats
(81.67±10.14%) but showed no significant
difference (p<0.05) in the sluggish sperm cell
levelsthough it increased (18.33±1.66%) in the
Lead-acetate + LDE administered with C. lanatus
seed.
The total sperm count was significantly (p<0.05)
higher in the Lead-acetate + HDE administered
rats (33.67±2.96X10
6
/ml) compared to the Lead-
acetate administered rats only (7.00±2.08
X10
6
/ml). The level of Sperm morphology (%) in
both Head, Mid-piece and Tail defect increased
significantly (p<0.05) in the Lead-acetate only
administered rats (compared 5.33 ± 2.40%, 2.33
±0.33% and 3.00±0.00% respectively) compared
to the Lead-acetate + HDE administered rats
(which had 0% Mid-piece defect, 0.66±0.33%
and 1.00±0.58% for Head and Tail defect
respectively).
Sperm viability (% viable) showed no significant
difference in either the Lead-acetate + HDE rats
Lead-acetate only rats or control rats. Though
the Sperm viability (% non-viable) of the Lead-
acetate only rats (10.67±2.33%) was significant
higher (p<0.05) as compared to the Lead-acetate
+ HDE albino male rats (1.67±.0.67%). The
Testosterone level of the Lead-acetate + HDE
rats (5.23±0.23ng/ml) was significantly higher
(p<0.05) compared to the Lead-acetate only rats
(0.73±0.12ng/ml).
Histological examination of rat testis in group 1
(control) as shown in Fig. 1 showed the
seminiferous tubules lined with stratified
epithelium composed of two major cells, which
are the sertoli cells and spermatogenic cells.
Seminiferous tubules depict normal architecture
with adequate cellularity; on the other hand,
histological examination in group 4 as shown in
Fig. 2 showed degeneration of the
spermatogenic cells, occlusion of the lumen and
hypertrophied seminiferous tubules. By contrast,
the lead acetate alone group depict shrunken
seminiferous tubules and reduced cellularity,
while group 2 shows increase in the
spermatogenic cells of the rats treated compared
with the control. This group showed a mild
thickening of the basement membrane as noted
in certain seminiferous tubules, but it appeared
generally healthy (Fig. 3). Finally, histological
examination of group 3 showed matured
spermatozoa in the seminiferous tubules of rats
treated. This group has a healthy testicular tissue
simila to the control, but with a slight increase in
the interstitial tissues. This shows that
methanolic extract of C. lanatus seed
administered as a treatment option may be
medicinally beneficial (Fig. 4).
4. DISCUSSION
Several studies show that alkaloids and terpenes
are widely spread in the genus citrullus [28].
These secondary metabolites are responsible for
the pharmacological activities such as antiulcer,
antimicrobial, antioxidant, analgesic, aphrodisiac
and many other ethno-medicinal uses [29].
Godspower et al.; BJPR, 6(1): 35-43, 2015; Article no.BJPR.2015.047
39
Table 3.1. Ameliorative potential of methanolic extracts of C. lanatus (watermelon) seeds on sperm parameters and testosterone level of lead-acetate induced male albino rats
Group
Sperm count (X10
6
/ml)
Sperm morphology (%)
Sperm viability (%)
Testosterone
(ng/ml)
AC
SC
DC
Sperm Count
HD
MPD
TD
VC
NVC
Testosterone
Control(1)
82.50±2.50 7.50±2.50 10.00±0.00 43. 50±1.50 0.50±0.50 0.00±0.00 0.50±0.50 99.00±1.00 1.00±1.00 2.75±1.45
Lead-acetate + LDE(2)
43.33±6.00 18.33±1.66* 38.33±4.41 20.00±1.55 2.33±1.20 1.00±0.58 1.00±0.58 95.67±0.33 4.33±0.33 1.77±0.52
Lead-acetate + HDE(3)
65.00±2.88* 15.00±5.00 20.0±2.88 33.67±2.96* 0.66±0.33 0.00±0.00 1.00±0.58 98.33±0.67 1.67±0.67 5.23±0.23*
Lead-acetate only(4)
11.66±7.26
6.66±3.33 81.66±10.14* 7.00±2.08 5.33±2.40* 2.33±0.33* 3.00±0.00* 89.33±2.33 10.67±2.33* 0.73±0.12
Data represented as Mean ± SEM; (*) p<0.05 significant difference; LDE=Low dose extract of C. lanatus seeds, HDE=High dose extract of C. lanatus seed; AC: Active cells; SC: Sluggish cells; Dead cells; HD: Head
defect; MPD: Mid-piece defect; TD: Tail defect; VC: Viable cells; NVC: Non-viable cells.
Godspower et al.; BJPR, 6(1): 35-43, 2015; Article no.BJPR.2015.047
40
Fig. 1. Photomicrograph of albino rat testis in group 1
Fig. 2. Photomicrograph of albino rat testis in group 4
Fig. 3. Photomicrograph of albino rat testis in group 2
Godspower et al.; BJPR, 6(1): 35-43, 2015; Article no.BJPR.2015.047
41
Fig. 4. Photomicrograph of albino rat testis in group 3
This study indicates that even moderate
exposure to Lead-acetate can significantly
reduce semen parameters, reduce testosterone
level and cause toxicity to the testis. The
changes observed in the above agree with the
previous reports, which demonstrated that lead
acetate suppressed testosterone, follicle-
stimulating hormone (FSH) and luteinizing
hormone (LH) levels along with testicular
spermatogenesis, showing that lead acts at all
levels of reproduction [30]. The extract
administered resulted to improvement of sperm
characteristics; this may be as a result of the
presence of flavonoids.The presence of steroid in
the phytochemical screening of the crude extract
could be the resultant improved testosterone
level in group 3. The mechanisms by which
methanolic extract of C. lanatus seeds protect
against experimentally induced testicular toxicity
may be as a result of the rich source of vitamin
C, thiamine and including riboflavin which
contains a high level of polyphenolic compounds
present in the plant. High concentration of
vitamin C in C. lanatus seeds provides highly
effective anti-oxidants, reversing the negative
effect caused by the lead-acetate following the
administration of the extracts to the experimental
animal as seen in the group III of the different
parameters when compared to the group 4. This
effect may be influenced by the presence of
flavonoids in the extract which contains
antioxidants. Saponin that was found to be
present in the extract functions majorly at
stimulating an increase in the body’s natural
endogenous testosterone levels which helps to
maintain testosterone levels. C. lanatus seeds
are a rich source of flavonoids and phenol, with
ability to scavenge free radical and inhibit
hydrolytic and oxidative enzymes and anti-
inflammatory action [31]. However, findings from
these results are supported by the fact that
methanolic extracts of C. lanatus seeds contains
a high amount of anti-oxidant activity [32],
bringing about the promising result obtained in
this study.
5. CONCLUSION
Methanolic extracts of C. lanatus seeds has
shown to be a beneficial treatment option against
lead-acetate induced oxidative stress and toxicity
in testicular tissue. It was also deduced from the
study that among the treatment options,
administration of methanolic extracts of C.
lanatus seeds at a high dose on exposure to lead
result to the most beneficial result such as
increased sperm motility, well defined cellularity
of the testis, increased sperm viability, decreased
sperm morphological alterations, increased
sperm count, increased testosterone level. There
is need for further investigations because
methanolic extracts of C. lanatus seeds can be a
potential complimentary agent in treating lead
induced testicular toxicity.
CONSENT
It is not applicable.
ETHICAL APPROVAL
All authors hereby declare that principle of
laboratory animal care was dully followed, as well
as specific national laws where applicable in this
study.
Godspower et al.; BJPR, 6(1): 35-43, 2015; Article no.BJPR.2015.047
42
COMPETING INTERESTS
Authors have declared that no competing
interests exist.
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... A regular watermelon juice consumption result in significant increases in blood plasma concentrations of lycopene and b carotene [8][9][10]. ...
... A six weeks study found that treatment with a C. lanatus extract containing 6 g of L-citrulline and L-arginine daily on middle-aged obese subjects with prehypertension or stage one hypertension experienced reduced ankle blood pressure and altered carotid wave reflection, an indication of improved arterial function of the individuals [10][11][12] . It is very important to note that all parts of the watermelon have something to offer. ...
... After 30 days of treatment the animals were fasted for 24 h prior to sacrifice. The animals were anesthetized using chloroform and then sacrificed [10]. Thus, blood collected via cardiac punctured and put in a labeled Ethylenediaminetetraacetic acid bottle (EDTA) for testosterone hormonal assay and later centrifuged at 7 000 rpm for ten (10) minutes. ...
... The protective effect of water melon seeds extract against MSG-induced testicular injury in male rats, reported in this study agree with that reported by [54], who concluded that, the extract of water melon seeds has ameliorative potentials on male reproductive system by increased sperm motility, well defined cellularity of the testis, increased sperm viability, decreased sperm morphological alterations, increased sperm count and increased testosterone level. Furthermore, the results of Khaki et al. [55] showed increase in sperm viability, motility and population of male rats received water melon seeds extract for 4 weeks so they concluded that it has a positive effect on male infertility. ...
... Moreover, Saponin that was found to be present in the seed extracts functions majorly at stimulating an increase in the body's natural endogenous testosterone levels which helps to maintain testosterone levels [54]. Also, Astaxanthin, a very potent antioxidant was reported to have positive effects on the reproductive system and particularly on infertility. ...
Article
Aims: Monosodium glutamate (MSG) is extensively used as food additive and flavor enhancer, there is a growing concern that this may affect the male reproductive system and fertility. The objective of this study is to investigate the effect of MSG on fertility and testes of mature male rats and the ameliorative role of water melon and cantaloupe (seeds extract and juices). Study Design: Thirty-six male Sprague - Dawely rats (150-180 g) were randomly assigned into six groups (n=6). Group (1): orally administered with distilled water. Group (2): orally administered with 60 mg/kg of MSG. Groups (3 and 4): orally administered with 60 mg/kg of MSG + 200 mg/kg of water melon seeds extract and juice respectively. Groups (5 and 6): orally administered with 60 mg/kg of MSG + 200 mg/kg of cantaloupe seeds extract and juice respectively. Results: Results showed that administration of MSG for 6 weeks caused abnormalities of semen characteristics, increased DNA damage and up-regulation of caspase3 expression in the testes tissue. Also, the levels of plasma sex hormones were decreased and the oxidant-antioxidant status was disturbed, moreover, MSG caused alteration in the histopathological structures of testicular tissue. Administration of seeds extract or juices of water melon and cantaloupe almost corrected the biochemical and histopathological alteration produced by MSG. Conclusion: This study concluded that water melon and cantaloupe seeds and juice extracts have an ameliorative role against MSG-induced testicular damage and infertility in rats.
... After 14 days of treatment, the experimental animals were fasted for 24 hours prior to sacrifice. The animals were anaesthetized using chloroform and then sacrificed [33]. ...
... Thus, blood collected via cardiac punctured and put in a labeled Ethylenediamminetetraaetic acid (EDTA) bottle for lipid profile and liver enzymes and later centrifuged at 7000 rpm for ten minutes [33]. The serum was then collected and stored at -15 0 C. The animals were then dissected; the pancreases were removed. ...
... Others are: antimicrobial activities, antigiardial activity and anti-inflammatory activity [30,31]. Methanolic extracts of Citrullus lanatus seeds has proven to be a treatment option against lead-acetate induced oxidative stress and toxicity in testicular tissue [32]. ...
... The degenerated interstitial space and congested seminiferous tubules seen in Group B was ameliorated in Group C possibly due to Citrullus lanatus administered which contains lycopene. Earlier findings supported this observation [32,17]. ...
... isms. Earlier studies have demonstrated that lead can pass through the blood testes barrier, accumulate in the testis and/or epididymis and seriously affect the spermatogonia, primary spermatocytes, spermatids or spermatozoa (germinal cells different levels of differentiation) [14]. ...
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Residents of the oil rich city of Port Harcourt and its environs have been inundated since November 2016 by a fresh wave of pollution plumes of thick black soot in the air capable of impacting health and wellbeing of the people. The current study investigated the relationship between exposure to environmental black soot and some reproductive and testicular function parameters in male Wistar rats. The study consisted of four groups of rats that were exposed to drinking water alone, while the remaining three groups were exposed to soot and sacrificed after four weeks, eight weeks and twelve weeks. Additionally, the control was also sacrificed at the end of weeks four, eight and twelve respectively. Results obtained indicated a statistically significant decrease (p < 0.05) in some sperm parameters and male sex hormones (follicle stimulating hormone, luteinizing hormone and testosterone) especially between the control and the group exposed to black soot for up to twelve weeks. Taken together, environmental exposure to black soot may alter hormonal profile and subsequently lead to infertility in exposed male rats.
... [27][28][29] The extract of CL pulp has shown ameliorative effects against monosodium glutamate-induced alterations in sperm motility, viability, and count, as well as altered sperm morphology in the rat. [30,31] Further, the beneficial role of CL rind methanolic extract on altered semen parameters and reproductive hormones against nicotine, [32] phenyl hydrazine, [33] and lead acetate [34] -induced toxicity has been reported in the rat. Another study suggested that fruit extract of CL provides protection to the sperm cells against arsenic-induced oxidative stress in the rat. ...
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Synthetic antidepressants are used in the treatment of mental illness. However, due to their side effects, people are orienting towards the herbal product as an alternative therapy. The present study, therefore, explores the role of Citrullus lanatus (CL) seed extract in evaluating the fluoxetine-induced impaired sperm functions and fertility of depressed mice. Adult Swiss strain male mice were separated into six groups. Every group contains six animals. Group I received vehicle-treated negative control while groups II, III, IV, V and VI were administered reserpine (RES) at a dose of 0.75 mg/kgBW/day, for 14 days to induce depression, fluoxetine (FLX) at the dose of 40 mg/kgBW/day in depressed mice (positive control) for 28 days, CL seed extract (300 mg/kgBW/day) only, in normal mice, and co-administrations of FLX (40 mg/kgBW/day) with low and high doses of seed extract of CL (150 and 300 mg/kg BW/day), respectively, in the depressed mice for the same duration. Mice of Gr. II and Gr. IV did not alter the epididymal sperm parameters and fertility. However, significant alterations were observed in such parameters in the mice of Gr. III. Partial to complete restorations were noticed in such parameters in Gr. V and VI, as compared to the positive control of Gr. III. The findings suggest that CL seed extract may emerge as a potential herbal remedy for impaired fertility, which could be of therapeutic drug in the future.
... The ameliorative potential of methanolic extract of CL seeds on lead acetateinduced testicular toxicity has been reported in the adult albino rat [80]. These authors have reported that administration of methanolic extract of CL seeds at a dose of 200mg/kg for 4 weeks in lead acetate-induced rat resulted in well-defined cellularity of the testis, an increased sperm motility, viability, and count, as well as the testosterone level, with reduced altered sperm morphology. ...
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In perspective of lifestyle that we are thriving in, has given rise to uncountable forms of diseases that seek medical assistance via utilization of synthetic drugs, resulting in certain side effects. These effects can be vanquished by using natural products as medicine. There exists a diverse range of flora that retains medicinal values and is majorly used by us in the form of food. One of such significant fruit crop is Citrullus lanatus which is known for its immense therapeutic and pharmacological properties. The present chapter provides the outstanding properties of every part of Citrullus lanatus that have been accomplished previously. The perception of using natural products over synthetic ones is due to their less or no side effects, cost-effectiveness and easy availability. Presence of several phytoconstituents such as alkaloids, flavonoids, tannins, amino acids, carbohydrates, glycosides, terpenoids, steroids, carotenoids, oils and fats can be effectively utilized in the treatment of various types of diseases such as cardiovascular, neuro-degenerative, diabetes, infertility, ulcer etc. Further, the nutrients present in various parts of Citrullus lanatus vitamin C, minerals, proteins, glycoprotein, and carbohydrates may be used as supplement against various health related issues. The objective of the present chapter is therefore to provide consolidated and authentic informations regarding the pharmacological properties of Citrullus lanatus based on experimental studies reported so far. This chapter has been formulated after going through several research articles, reviews, book chapters etc. that are indexed in platforms namely google scholar, scopus, DOI and Elsevier. Keywords: Citrullus lanatus; natural products; pharmacological properties.
... Male infertility account for 20-30% of infertility cases and it is characterised by semen quality deterioration, which culminates into low sperm cell counts, abnormal sperm morphology as well as hormonal imbalance (Godspower et al., 2015;Dissanayake et al., 2019). Since the rabbit is an excellent human model (Esteves et al., 2018), it is suggestive that soursop juice enhances spermatozoa quality and mitigates lipid peroxidation by improving antioxidant capacity in male animals. ...
Article
Heat induced stress associated with dry climatic conditions in the tropics does have adverse effects reproduction in rabbits, and this in-turn impacts negatively on the income of rabbit farmers. However, natural products might prove to be a reliable, safe and cheap remedy for ameliorating reproductive such anomalies in rabbits. The potential of soursop for mitigating heat induced reproductive deficiency in rabbit bucks was investigated during the peak of dry climatic conditions in southwestern Nigeria. Sixty mixed breed (New Zealand white x Chinchilla) adult rabbit bucks were allotted to five treatments of four replicates (3 bucks per replicate) each in a completely randomized design. Soursop (Annona muricata) fruit was processed into juice using standard procedures and was designated as soursop juice. The juice was administered orally daily per kg body weight (BW), 0.55ml/kgBW distilled water (control), 0.55 ml/kgBW soursop juice, 1.11 ml/kgBW soursop juice, 1.67 ml/kgBW soursop juice and 2.22 ml/kgBW soursop juice to designated treatments 1 to 5, respectively for 56 days. Semen samples were collected with an improvised artificial vagina on the 28th and 56th day of the study, semen quality and seminal oxidative status were evaluated using standard procedures. Results showed that rabbit buck exposed to heat stress had lower semen quality, seminal antioxidants and increased seminal lipid peroxidation. However, the consumption of soursop juice lowered lipid peroxidation and enhanced (p < 0.05) antioxidant production in the seminal fluid of heat-stressed bucks than bucks on control group. Bucks' semen quality and antioxidant status peaks in heat-stressed bucks gavaged 2.22 ml/kgBW soursop juice and gives 100% recovery from the effects of heat induced stress. It can be concluded that 2.22 ml/kgBW soursop juice administered for 56 days did enhance spermatozoa quality and mitigated lipid peroxidation by improving antioxidant capacity of male rabbits in a dose dependent manner on extremely dry climatic conditions.
... Also, decrease levels of IL 6 were observed in mice with adipose tissue inflammation when treated with lycopene (Gouranton et al. 2011). The mechanisms by which W J protect against -irradiation induced testicular oxidative damage may be as a result of the rich source of vitamin C, thiamine and including riboflavin which contains a high level of polyphenolic compounds present in this fruit (Godspower et al. 2015). W J is an excellent source of lycopene, having about 40% higher lycopene content than raw tomatoes (Seif, 2014). ...
Article
Background: The present study was designed to evaluate the expected beneficial effect of watermelon juice (WJ) on γ-radiation induced inflammation and testes tissue damage in rats.Methods: The values for total phenolic contents, antioxidant activity and 2,2-diphenyl-1-picrylhydrazyl for WJ were detected. 28 Male rats were randomly divided into 4 groups (seven rats in each group) as follows; Control group, WJ group (received WJ by oral gavage 100 mg/kg B.Wt. / day/ 6 weeks), group 3 and 4: Irradiated group only (6Gy; the 1st week) and irradiated and WJ group, respectively. At the end of the experiment, blood and tissue samples were collected for biochemical analysis and histopathological examination.Result: It has been found that γ-irradiation led to testicular oxidative stress accompanied by hormonal disturbance (elevation of follicle stimulating hormone (FSH) and reduction of testosterone and luteinizing hormone (LH) levels) and elevation of inflammatory factors (serum tumor necrotic factor-alpha (TNF-α), interleukin-6 (IL-6) ). Rats received WJ after exposure to γ-rays showed significantly less severe damage and remarkable improvement in all of the measured parameters and also restored the histological changes in the testis when compared to irradiated rats.
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Lead acetate, a prominent heavy metal, is released into the environment by various industries like paint, ceramics, lead-containing pipes, and plastics. Exposure to lead acetate has adverse effects on numerous organs in the body, with a particular impact on the testes due to its distinctive vascular system. The present study investigated the potential protective effects of biotin on lead acetate-induced testicular damage in Wistar rats. Twenty (20) male Wistar rats were divided into 4 groups: control (animal feed daily with water); negative control (lead acetate 60 mg/kg); Positive control group( high-dose Biotin (80mg/kg) and the treatment group Lead acetate (60mg/kg)/high-dose Biotin (80mg/kg). After 28 days of administration, blood samples were collected for hormonal assay, and semen from the epididymis for semen profile. Testicular samples were also collected for histopathological studies. Results showed that lead acetate administration significantly decreased the sperm count, motility, viability, and altered histology of the testis (testicular damage, necrosis of seminiferous tubules, and loss of spermatid) compared to the negative control. However, the treatment group showed significantly improved histology of the testis, and increased sperm count, motility, and viability. From the results of this study, it could be concluded that biotin supplementation could provide a promising ameliorative effect against lead acetate-induced testicular toxicity.
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Ficus religiosa is being used in Ayurvedic and Malay traditional medicine for the treatment of various diseases including gastric ulcer. Considering the above claims, the present work was undertaken to validate the anti-ulcer potential of the ethanol extract of stem bark of F. religiosa against in vivo indomethacin- and cold restrained stress-induced gastric ulcer, and pylorus ligation assays. The extract (100, 200 and 400 mg/kg) significantly (P<0.05) reduced the ulcer index in all assays used. The extract also significantly (P<0.05) and increased the pH of gastric acid while at the same time reduced the volume of gastric juice and, free and total acidities. In conclusion, the present study provide preliminary data on the antiulcer potential of F. religiosa stem bark and support the traditional uses of the plant for the treatment of gastric ulcer.
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The carotenoid content and in vitro accessibility of the 'Sugar baby' variety of watermelon and the 'Horana red' variety of guava from Sri Lanka was determined using high-performance liquid chromatography. The high-performance liquid chromatography chromatogram showed that the Guava 'Horana red' variety contained almost exclusively lycopene (45.3+/-8.0microg/g fresh weight (FW)), with a small amount of lutein (2.1+/-0.6microg/g FW), beta-carotene (2.0+/-0.2microg/g FW) and beta-cryptoxanthin. As far as carotenoids in the sugar baby variety of watermelon are concerned, it contained lycopene, lutein and beta-carotene of 37.2+/-4.0microg/g FW, 2.1+/-0.6microg/g FW and 0.3+/-1microg/g FW, respectively. The studies showed that guava contains more lycopene (45.3+/-8.0microg/g FW) than watermelon (37.2+/-4.0microg/g FW), and that the in vitro accessibility of lycopene in guava (73%) is more than that in watermelon (25.8%). Therefore it can be concluded that guava can be used as a better lycopene source than watermelon.
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Background: Medical nutrition therapy is recognized as an important treatment option in type 2 diabetes. Most guidelines recommend eating a diet with a high intake of fiber-rich food including fruit. This is based on the many positive effects of fruit on human health. However some health professionals have concerns that fruit intake has a negative impact on glycemic control and therefore recommend restricting the fruit intake. We found no studies addressing this important clinical question. The objective was to investigate whether an advice to reduce the intake of fruit to patients with type 2 diabetes affects HbA1c, bodyweight, waist circumference and fruit intake. Methods: This was an open randomized controlled trial with two parallel groups. The primary outcome was a change in HbA1c during 12 weeks of intervention. Participants were randomized to one of two interventions; medical nutrition therapy + advice to consume at least two pieces of fruit a day (high-fruit) or medical nutrition therapy + advice to consume no more than two pieces of fruit a day (low-fruit). All participants had two consultations with a registered dietitian. Fruit intake was self-reported using 3-day fruit records and dietary recalls. All assessments were made by the "intention to treat" principle. Results: The study population consisted of 63 men and women with newly diagnosed type 2 diabetes. All patients completed the trial. The high-fruit group increased fruit intake with 125 grams (CI 95%; 78 to 172) and the low-fruit group reduced intake with 51 grams (CI 95%; -18 to -83). HbA1c decreased in both groups with no difference between the groups (diff.: 0.19%, CI 95%; -0.23 to 0.62). Both groups reduced body weight and waist circumference, however there was no difference between the groups. Conclusions: A recommendation to reduce fruit intake as part of standard medical nutrition therapy in overweight patients with newly diagnosed type 2 diabetes resulted in eating less fruit. It had however no effect on HbA1c, weight loss or waist circumference. We recommend that the intake of fruit should not be restricted in patients with type 2 diabetes. Trial registration: http://www.clinicaltrials.gov; Identifier: NCT01010594.
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The mutagenic and carcinogenic potentials of lead are still being investigated. The aim of this study was to evaluate the genotoxic effect of lead acetate in the early period of life, when the organism is extremely sensitive to toxic effects of lead. Six-day-old suckling Wistar rats were exposed to lead (as acetate) either orally for 9 days (daily dose 2 mg Pb/kg b. wt., 18 mg/kg b.wt. total dose) or by a single intraperitoneal injection (5 mg Pb/kg b. wt.). DNA damage was investigated using the comet assay and in vivo micronucleus test. The results of the comet assay showed statistically significant differences between the control (unexposed) animals and the two groups of exposed animals by ANOVA weighted for unequal variance (heterogeneity of variances was found by Levene's test), followed by Tukey's post hoc test at the level of significance of p < 0.05. The two groups of lead-exposed animals were also significantly different from each other. Orally lead-exposed animals showed a significant increase of micronuclei frequencies in reticulocytes and erythrocytes compared to unexposed animals (ANOVA, p < 0.05).
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Total phenolic content (TPC) and ascorbic acid equivalent antioxidant capacity (AEAC) of leaves of 26 ginger species belonging to nine genera and three tribes were screened. For 14 species, TPC and AEAC of rhizomes were also assessed. Ferrous ion-chelating (FIC) abilities of leaves and rhizomes of eight species were compared. Leaves of five species of Etlingera were analysed for tyrosinase inhibition activity. Of the 26 species, leaves of Etlingera species had the highest TPC and AEAC. Eleven of the 14 species had significantly higher TPC and/or AEAC in leaves than in rhizomes. Values of leaves of Etlingera elatior and Etlingera maingayi were seven to eight times higher than those of rhizomes. In terms of FIC ability, six of the eight species clearly showed higher values in leaves than in rhizomes. The most outstanding was the FIC value of Alpinia galanga leaves which was more than 20 times higher than that of rhizomes. Of the five species of Etlingera, leaves of E. elatior displayed the strongest tyrosinase inhibition activity, followed by leaves of Etlingera fulgens and E. maingayi. Values of their inhibition activity were significantly higher than or comparable to the positive control. Besides promising tyrosinase inhibition ability, leaves of these three Etlingera species also have high antioxidant activity and antibacterial properties.
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Ankle-brachial index (ABI) and ankle blood pressure (BP) are associated with increased carotid wave reflection (augmentation index, AIx). Oral L-citrulline and L-arginine from synthetic or watermelon sources have reduced brachial BP, aortic BP, and aortic AIx. A directly measured carotid AIx (cAIx) rather than aortic AIx has been proposed as a better measurement of central AIx. We evaluated the effects of watermelon extract on ankle BP and cAIx in individuals with normal ABI and prehypertension or stage 1 hypertension. Ankle and brachial systolic BP (SBP), diastolic BP (DBP), mean arterial pressure (MAP), cAIx, ABI, and heart rate (HR) were evaluated in the supine position in 14 adults (11 women/3 men, age 58 ± 1 years) with prehypertension or stage 1 hypertension (153 ± 4 mm Hg). Subjects were randomly assigned to 6 weeks of watermelon extract supplementation (L-citrulline/L-arginine, 6 g daily) or placebo followed by a 2-week washout period and then crossover. Ankle and brachial SBP (-11.5 ± 3.8 and -15.1 ± 2.8 mm Hg), DBP (-7.8 ± 2.3 and -7.6 ± 1.8 mm Hg), and MAP (-9.8 ± 2.6 and -7.3 ± 1.8 mm Hg), and cAIx (-8.8 ± 2.6 %) decreased significantly (P < 0.05) after watermelon supplementation compared to placebo. Watermelon supplementation had no significant effect (P > 0.05) on ABI and HR. This study shows that watermelon extract supplementation reduces ankle BP, brachial BP, and carotid wave reflection in obese middle-aged adults with prehypertension or stage 1 hypertension and normal ABI, which may reflect improved arterial function.
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Adverse effects of chemical pollutants on spermatogenesis have become a relevant issue in modern andrology, since environmental chemical contamination increases everyday world-wide. Agropesticides are one important group of such chemicals. Their relevance is due to their widespread use, mostly in the Third World, and to the various ways they interfere with male reproduction. Most organochlorine pesticides behave as xenoestrogens and though they have been banned in most countries many years ago, they have long lasting effect in the biosphere and are still illegally used. They have been largely replaced by organophosphorous pesticides (OP) which since then have been demonstrated as hazardous to human reproduction. This article provides a brief review on agropesticides, their use in Latin America, and occupational and environmental risks. Some of the clinical and biological effects of an OP pesticide parathion are presented. Animal experiments showed that OP chemicals damage germ and somatic cells of the testis, both at the onset of spermatogenesis and in the adults. Effects are not only genotoxic, but also teratogenic. The present knowledge advices prompt replacement of chemical pest control by environmentally safer methods.
Article
Adverse effects of chemical pollutants on spermatogenesis have become a relevant issue in modern andrology, since environmental chemical contamination increases everyday world‐wide. Agropesticides are one important group of such chemicals. Their relevance is due to their widespread use, mostly in the Third World, and to the various ways they interfere with male reproduction. Most organochlorine pesticides behave as xenoestrogens and though they have been banned in most countries many years ago, they have long lasting effect in the biosphere and are still illegally used. They have been largely replaced by organophosphorous pesticides (OP) which since then have been demonstrated as hazardous to human reproduction. This article provides a brief review on agropesticides, their use in Latin America, and occupational and environmental risks. Some of the clinical and biological effects of an OP pesticide parathion are presented. Animal experiments showed that OP chemicals damage germ and somatic cells of the testis, both at the onset of spermatogenesis and in the adults. Effects are not only genotoxic, but also teratogenic. The present knowledge advices prompt replacement of chemical pest control by environmentally safer methods.
Article
People in developing countries depend largely on non-conventional protein sources to augment the availability of proteins in their diets. Watermelon seed meal is reported to contain an adequate amount of nutritional proteins that could be extracted for use as nutritional ingredients in food products. Osborne classification showed that globulin was the major protein (≥500 g kg (-1)) present in watermelon seed meal, followed by albumin and glutelin. Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that the polypeptides had low molecular weights ranging from 35 to 47 kDa. Isoelectric focusing revealed that the isoelectric point of most proteins was in the acidic range 4-6. These proteins are rich in aspartic acid, glutamic acid and serine. An increase in pH (5-9) significantly (P < 0.05) decreased the denaturation enthalpy of these proteins. Among functional properties, albumin exhibited a much higher dispersibility index (810.3-869.6 g kg(-1)) than globulin (227.8-245.4 g kg(-1)), glutelin (182.1-187.7 g kg(-1)) and prolamin (162.3-177.7 g kg(-1)). Digestibility was in the ranges 760.6-910.0 and 765.5-888.5 g kg(-1) for Mateera and Sugar Baby watermelon protein fractions respectively, while surface hydrophobicity ranged from 126.4 to 173.2 and from 125.8 to 169.3 respectively. The foaming and emulsifying properties of albumin were better than those of the other proteins studied. The good nutritional and functional properties of watermelon seed meal proteins suggest their potential use in food formulations.
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Lead is a ubiquitous toxic heavy metal with unique physical and chemical properties that make it suitable for a great variety of applications. Because of its high persistence in the environment and its use since ancient times for many industrial activities, lead is a common environmental and occupational contaminant widely distributed around the world. Even though the toxic effects of lead and its compounds have been investigated for many years in a variety of systems, the data existing with regard to its mutagenic, clastogenic and carcinogenic properties are still contradictory. The International Agency for Research on Cancer has classified lead as possible human carcinogen (group 2B) and its inorganic compounds as probable human carcinogens (group 2A). Furthermore, although the biochemical and molecular mechanisms of action of lead remain still unclear, there are some studies that point out indirect mechanisms of genotoxicity such as inhibition of DNA repair or production of free radicals. This article reviews the works listed in the literature that use different parameters to evaluate the genotoxic effects of lead in vitro, in vivo and in epidemiological studies.