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The present investigation was undertaken to assess the role of Mucuna pruriens in infertile men who were under psychological stress. Study included 60 subjects who were undergoing infertility screening and were found to be suffering from psychological stress, assessed on the basis of a questionnaire and elevated serum cortisol levels. Age-matched 60 healthy men having normal semen parameters and who had previously initiated at least one pregnancy were included as controls. Infertile subjects were administered with M. pruriens seed powder (5 g day(-1)) orally. For carrying out morphological and biochemical analysis, semen samples were collected twice, first before starting treatment and second after 3 months of treatment. The results demonstrated decreased sperm count and motility in subjects who were under psychological stress. Moreover, serum cortisol and seminal plasma lipid peroxide levels were also found elevated along with decreased seminal plasma glutathione (GSH) and ascorbic acid contents and reduced superoxide dismutase (SOD) and catalase activity. Treatment with M. pruriens significantly ameliorated psychological stress and seminal plasma lipid peroxide levels along with improved sperm count and motility. Treatment also restored the levels of SOD, catalase, GSH and ascorbic acid in seminal plasma of infertile men. On the basis of results of the present study, it may be concluded that M. pruriens not only reactivates the anti-oxidant defense system of infertile men but it also helps in the management of stress and improves semen quality.
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Advance Access Publication 18 December 2007 eCAM 2010;7(1)137–144
Original Article
Mucuna pruriens Reduces Stress and Improves
the Quality of Semen in Infertile Men
Kamla Kant Shukla
, Abbas Ali Mahdi
, Mohammad Kaleem Ahmad
Shyam Pyari Jaiswar
, Satya Narain Shankwar
and Sarvada Chandra Tiwari
Department of Biochemistry,
Department of Obstetric & Gynaecology,
Department of Urology and
Department of Psychiatry, King George’s Medical University, Lucknow 226003, India
The present investigation was undertaken to assess the role of Mucuna pruriens in infertile men
who were under psychological stress. Study included 60 subjects who were undergoing infertility
screening and were found to be suffering from psychological stress, assessed on the basis of a
questionnaire and elevated serum cortisol levels. Age-matched 60 healthy men having normal
semen parameters and who had previously initiated at least one pregnancy were included as
controls. Infertile subjects were administered with M. pruriens seed powder (5 g day
) orally.
For carrying out morphological and biochemical analysis, semen samples were collected twice,
first before starting treatment and second after 3 months of treatment. The results
demonstrated decreased sperm count and motility in subjects who were under psychological
stress. Moreover, serum cortisol and seminal plasma lipid peroxide levels were also found
elevated along with decreased seminal plasma glutathione (GSH) and ascorbic acid contents
and reduced superoxide dismutase (SOD) and catalase activity. Treatment with M. pruriens
significantly ameliorated psychological stress and seminal plasma lipid peroxide levels along
with improved sperm count and motility. Treatment also restored the levels of SOD, catalase,
GSH and ascorbic acid in seminal plasma of infertile men. On the basis of results of the present
study, it may be concluded that M. pruriens not only reactivates the anti-oxidant defense system
of infertile men but it also helps in the management of stress and improves semen quality.
Keywords: antioxidants – lipid peroxides – male infertility – Mucuna pruriens – psychological
Infertility is a major public health concern and it is said to
be the manifestation of one or more pathological condi-
tions of either male or female origin. As many as 15% of
couples have difficulty in conceiving, and the male factor is
implicated as the cause in up to 50% of such cases.
Moreover, in 10–15% of infertile couples, no apparent
cause can be found and these cases are categorized under
‘unexplained infertility’ (1). The problem of infertility is
closely related to stress, as a couple, failing to achieve the
expected goal of reproduction, experiences the feelings of
frustration and disappointment. These feelings only
compound in couples experiencing infertility related
problems requiring prolonged efforts to conceive.
Previous studies have indicated that stress, especially
psychological stress, has a negative impact on various
parameters associated with semen quality, including sperm
concentration, motility and morphology. Other distur-
bances, such as impotence, sham ejaculation, retrograde
ejaculation and oligospermia, have also been reported
to be associated with psychological factors underlying
male infertility (2).
For reprints and all correspondence: Prof. Dr Abbas Ali Mahdi,
Department of Biochemistry, King George’s Medical University,
Lucknow 226003, India. Tel: +91-9839011192; Fax: +91-522-2257539;
ß2007 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (
licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original workis
properly cited.
Chronic exposure to psychological stress is also known
to cause a variety of patho-physiological changes in
neuroendrocrine system, resulting in altered steroidogen-
esis and spermatogenesis. Prolonged stress also leads to
increased blood cortisol level. An excess of this hormone,
in turn, markedly affects the spermatogenesis, resulting in
retention of cytoplasmic droplets and over-production of
immature spermatozoa, which are reported to be the
major source of reactive oxygen species (ROS) (3).
Moreover, there are also reports that elevated psycholog-
ical stress itself is associated with increased oxidant
production and long-term exposure to psychological
stressors may enhance the generation of ROS (4). Under
normal circumstances, the ROS scavenging potential of
the male reproductive tract and seminal fluid is maintained
by adequate levels of enzymatic and non-enzymatic
antioxidants, vitamins and minerals (5,6). On the other
hand, when the production of ROS is high, it may lead to
oxidative damage to spermatozoa. ROS is also reported to
have a negative effect on sperm functioning. The sperm
plasma membrane is very sensitive to ROS, since it
contains high levels of unsaturated fatty acids. The latter
provide fluidity, which is necessary for sperm motility and
acrosome reaction. Increased ROS level, as found during
prolonged psychological stress, may lead to an imbalance
between oxidant/anti-oxidant ratio, leading to increased
lipid peroxidation, resulting in sperm membrane damage
and its subsequent dysfunctioning (7).
Till date, appropriate treatment of idiopathic male
infertility has not been found. However, in the ancient
Indian system of medicine, the Ayurveda and Unani, as
also the Chinese system of traditional remedies, some
plants e.g. Mucuna pruriens,Tinospora cordiofolia,
Asparagus racemosus,Withania somnifera,Orchis latifolia,
Ocimum basilium and Tribulus terrestris etc. were used for
the improvement of endurance against stress, general
resistance against infections, retardation of the aging
process and improving male sexual disorders, like
psychogenic impotence and unexplained infertility (8–10).
Mucuna pruriens Linn. (Fabaceae), commonly known
as ‘cowhage plant’ or ‘kapikacho’ or ‘kevach’ in Hindi, is
the most popular drug in the Ayurvedic and Unani
system of medicine. Its different preparations (from the
seeds) are used for the management of several free
radical-mediated diseases, such as rheumatoid arthritis,
diabetes, atherosclerosis, nervous disorders and male
infertility (11). There are reports that seed powder of
M. pruriens helps in some way against stress, it increases
secretion of semen and it acts as a restorative and an
invigorating tonic or aphrodisiac in diseases characterized
by weakness or loss of sexual power (12). It is also used
in the management of Parkinsonism, as it is a good
source of L-3, 4 dihydroxyphenyl alanine (L-DOPA) (13).
Seeds of M. pruriens also possess antioxidant, hypogly-
cemic, lipid lowering and neuroprotective activities (14). Its
seeds contain the alkaloids, mucunine, mucunadine,
mucunadinine, prurienidine and nicotine, besides
b-sitosterol, glutathione, lecithin, vernolic acid and gallic
acid. They posses a number of other bioactive substances,
including tryptamine, alkylamines, steroids, flavonoids,
coumarins, cardenolides and metals like magnesium,
copper, zinc, manganese and iron (15). The present study
was planned to investigate the effect of M. pruriens on
semen quality and the seminal plasma levels of lipid
peroxides and antioxidants in infertile men, who were
under psychological stress.
Materials and Methods
Nitroblue tetrazolium Cat N-5514 (NBT), thiobarbituric
acid Cat T-5500 (TBA), phenazine methosulphate Cat
N-9625 (PMS), nicotinamide adenine dinucleotide Cat
N-6754 (NADH), 5.50-dithio bis 2- nitrobenzoic acid
Cat D-5420 (DTNB), 4,5 methyl thiazol-2-yl 2,5 diphenyl
tetra zolium bromide Cat M-2128 (MTT), nicotinamide
adenine dinucleotide phosphate Cat N-7785 (NADPH)
tricholoroacetic acid Cat T-8657 (TCA) and reduced
glutathione Cat G-4251 (GSH) were purchased from
Sigma Chemical Co., St Louis, MO, USA. Radioimmu-
noassay kit for estimation of serum cortisol was obtained
from Diagnostic Product Corporation, Los Angeles, CA,
USA. This kit with trade name Code-A-Count Cortisol,
contains cortisol antibody coated polypropylene tubes
(TC 01), a solution of iodinated cortisol
I cortisol
(TC 02) as well as cortisol calibrators (TC 03-8) containing
standard cortisol. All other reagents used were of high
quality and analytical grade.
Plant Material
The seeds of plant M. pruriens were purchased from an
authorized dealer in Lucknow, India. They were identified
and authenticated by Dr M.M.A.A Khan, Senior Lecturer,
Department of Botany, Shia P.G. College, Lucknow, India
(Herbarium No. M-113 dated October 17, 2005). The seeds
were dried under shade and made to a fine powder using
laboratory mill.
Study Protocol
The study protocol was approved by the Ethical
Committee of the King George’s Medical University,
Lucknow, (vide communication No.126/R-Cell-04, dated:
May 14, 2004). Before enrolment in the study, written
informed consent from each subject was obtained in
response to a fully written and verbal explanation of
the nature of the study. The potential participants, each
with infertility persisting longer than a year, were
clinically examined before being included in the study.
138 Mucuna pruriens in infertile men
Complete medical history of subjects and their female
partners was also recorded. Subjects having diabetes,
hypertension, arthritis, malignancy, tuberculosis, human
immunodeficiency virus positive, infections, endocrinal
disorders and on drugs and conditions known to influence
oxidative stress and serum cortisol level were excluded.
A total of 120 men, aged 30–38 years, belonging to the
same socioeconomic and ethnic status (Indo Aryan) and
body mass index ranging from 19 to 24 kg m
, were
selected from the couples attending the Infertility Clinic
of the Department of Obstetrics & Gynecology and the
Out Patient Department of Urology, King George’s
Medical University, Lucknow. All subjects were
instructed not to take any nutritional supplement or
vitamins and not to change their dietary habits during the
course of study. This study was undertaken from January
2006 to December 2006.
Sample Collection
Semen samples were collected from subjects with 3–4
days of sexual abstinence. Semen analysis was carried out
according to the World Health Organization guidelines
(16). Venous blood samples were also withdrawn and
serum separated for assessment of cortisol levels.
The prospective study included four parallel groups of
subjects: The control group comprised of 60 age-matched
healthy men who had previously initiated at least one
pregnancy and exhibited a normal semen profile
(>20 10
spermatozoa ml
; >40% motility and
>40% normal morphology). Moreover, control subjects
were also not under stress, as evidenced by the study
questionnaire and normal serum cortisol level.
The study group comprised of 60 subjects who were
under psychological stress (as assessed by the study
questionnaire and on the basis of elevated serum cortisol
levels). Study subjects were further divided into three
subgroups on the basis of semen profiles and in each
subgroup a minimum of 20 patients were included. These
groups were (i) normozoospermic infertile men, defined
as control group (ii), oligozoospermic infertile men
(<20 10
, spermatozoa ml
, motility >40% and
>40% normal morphology) and (iii) asthenozoospermic
infertile men (>20 10
spermatozoa ml
, <40% moti-
lity and >40% normal morphology).
Stress Questionnaire
Psychological stress was assessed as per the protocol
known as State Anxiety Inventory (17). All participants
were asked to complete the questionnaire of the State
Anxiety Inventory, as validated by Oner and Le Comple
(18). In the questionnaire, subjects were asked to describe
how they feel ‘right now’ by responding to 20 questions
with a 4-point response format from ‘not at all’ (score 1)
to ‘extremely’ (score 4). Total scores ranged from 20 to
80, with higher scores indicating greater anxiety. This
measure has been shown to have high reliability and
good contrast validity.
The infertile men were prescribed M. pruriens seed
powder (5 g day
), orally, in a single dose for 3
months with a cup of skimmed milk (19). Semen samples
were collected twice, i.e. first before administrating the
medicine and second, after 3 months of treatment. All the
semen morphological profiles were assessed within an
hour of sample collection and biochemical profiles were
evaluated within 2 days. During the course of study the
patients were also monitored, on monthly basis, for liver
Preparation of Seminal Plasma
Semen samples were collected by masturbation after 3–4
days of abstinence into sterile plastic containers for
analysis. The semen volume was recorded and an aliquot
was taken to assess sperm motility after at least 30 min
given for liquefaction. There after, the semen samples
were centrifuged at 1200 gin cold (4C) for 20 min
for the separation of seminal plasma. The supernatant
(seminal plasma) was again centrifuged at 10 000 gin
cold (4C) for 30 min to eliminate all possible contam-
inating cells and stored at 20C until analyzed.
All biochemical estimations were carried out in seminal
Biochemical Assay
Estimation of Lipid Peroxide Levels
Seminal plasma (0.2 ml) was mixed with 0.5 ml glacial
acetic acid. Subsequently, 0.5 ml of 8% sodium dodecyl
sulfate was added to the above reaction mixture. After
mixing well 1.5 ml of 0.8% TBA solution was added.
The reaction mixture was kept in boiling water bath for 1 h.
After cooling to room temperature, 3.0 ml of n-butanol
was mixed, the reaction mixture was then centrifuged
at 10 000 gfor 15 min. The absorbance of clear butanol
fraction obtained after centrifugation was read at
532 nm by UV-VIS double beam spectrophotometer
(model No. 2203, serial No. 053, Systronics Instrumental
Co. Hyderabad, India, year of purchase 2006). An
appropriate standard made up of 2.5 nmol malondialde-
hyde was run simultaneously (20).
Assay of Catalase Activity
Diluted hydrogen peroxide (0.2 ml of 20-fold dilution, 30%
w/v) and 2.5 ml of 50 mM phosphate buffer (pH 8.2) was
eCAM 2010;7(1) 139
added in a cuvatte. To the resultant, 0.02 ml of seminal
plasma, as enzyme source was added and mixed thor-
oughly. The decrease in the absorbance of reaction mixture
due to catabolism of hydrogen peroxide was recorded at
240 nm after every 30 s by spectrophotometer (21).
Assay of Superoxide Dismutase Activity
Two reaction setups were run in parallel for superoxide
dismutase (SOD) estimation. The tubes in the first setup
(experimental) received 0.2 ml (320 mM) NBT, 0.2 ml
(10 mM) phenazine methosulfate, 2.0 ml (0.16 mM) pyro-
phosphate buffer pH 9.2, 0.02 of seminal plasma as enzyme
source. The tubes in the second setup (reference) received
the entire above reagents except the enzyme source. The
reaction was started simultaneously in both sets by the
addition of 0.2 ml (160 mM) of NADH. After an interval of
90 s, 1 ml of glacial acidic acid was added to each reaction
tube. The reference tubes were then added with the same
amount of enzyme source, and absorbance was read at
560 nm against a blank on spectrophotometer (22).
Estimation of Reduced Glutathione
Seminal plasma 0.2 ml was mixed with 3.0 ml of 5% (w/v)
TCA reagent and allowed to stand for 5 min, proteins
were precipitated and filtered out. Later, 2.0 ml of filtrate
was added to 4.0 ml of 0.3 M phosphate buffer pH 7.4
and 1 ml of DTNB (1% w/v aqueous sodium citrate).
A blank was prepared in a similar manner using distilled
water in place of the filtrate. An appropriate standard
solution of 0.1 ml GSH (10 mmol) was also run simulta-
neous. The pale yellow color developed was read
immediately at 412 nm by spectrophotometer (23).
Estimation of Ascorbic Acid
Phosphate citrate buffer 1.5 ml (pH 8.2), 0.2 ml PMS,
0.2 ml MTT and 0.1 ml of seminal plasma were added in
a test tube and incubated at 37C for 15 min. The
reaction was stopped by adding 0.5 ml acetic acid. An
appropriate standard solution of ascorbic acid (0.1 ml)
was also run simultaneously. Brown color was developed
and measured by spectrophotometer at 578 nm (24).
Estimation of Serum Cortisol Levels
Serum cortisol levels were assessed by radioimmunoassay
method (PC-RIA-MAS, Gamma Counter, Stratec-
Germany, serial No. 2486000031 year of purchase-
2002) (25).
Statistical Analysis
Normal healthy fertile men and infertile groups were
compared using one-way ANOVA analysis of variance
followed by Dunnett test. Infertile groups, before and
after treatment, were compared with paired t-test. A
probability P-value of <0.05 (P< 0.05) was considered
statistically significant. The statistical analysis was
performed on commercial software INSTAT 3.0, a
demo version (Graph Pad Software, San Diego, CA).
Effect of M. pruriens on Semen Parameters
in Infertile Men
General semen characteristics of different groups of
subject, before and after treatment with M. pruriens, are
depicted in Fig. 1. In normal healthy fertile men (non-
stress, control group), the mean sperm concentration was
57.0 8.4 10
, motility was 57.0 5.4% and lique-
faction time was 21.5 2.5 min. The sperm concentration
and motility in all under stress infertile groups as compared
with controls were found decreased. The maximum
decrease in sperm concentration was observed in oligo-
zoospermic group (87%; P< 0.001) and motility of
spermatozoa in asthenozoospermic patients (78%;
P< 0.001). Treatment of these infertile males with
M. pruriens seed powder (5 g/day) for 3 months showed
significant reversal of the above findings. Our results show
that sperm concentration was most significantly increased
in oligozoospermic patients (+688%; P< 0.001) and
sperm motility was significantly increased in asthenozoos-
permic patients (+32%; P< 0.05).
Effect of M. pruriens on Stress Parameters
in Infertile Men
Stress scores, elaborated on the basis of the questionnaire,
were found significantly high in infertile subjects (Table 1).
Similarly, the morning serum cortisol levels were found
elevated in normozoospermic (+38%; P< 0.01), oligo-
zoospermic (+110%; P< 0.01) and asthenozoospermic
(+171%; P< 0.01) infertile men as compared with control
subjects (10.2 0.2 mgdl
). After treatment with
M. pruriens, serum cortisol levels were decreased in
normozoospermic (25%; P< 0.001), oligozoospermic
(81%; P< 0.001) and asthenozoospermic (55%;
P< 0.001) patients. Moreover, the mean serum cortisol
levels observed at 1600 h, in control group were
5.5 0.6 g dl
. But these levels in normozoospermic
(99%; P< 0.01), oligozoospermic (162%; P< 0.01) and
asthenozoospermic (231%; P< 0.01) were found to be
increased. After treatment with M. pruriens, significant
reversal of serum cortisol levels in normozoospermic
(40%; P< 0.001), oligozoospermic (46%; P< 0.001) and
asthenozoospermic (49%; P< 0.001) was observed.
The intra and inter assay variances of serum cortisol were
<6% and <10%, respectively, which was found to be
highly significant.
140 Mucuna pruriens in infertile men
Effect of M. pruriens on Lipid Peroxide Levels and
Antioxidant Parameters in Infertile Men
The lipid peroxide level in seminal plasma of control
healthy men was 2.2 0.3 nmol MDA ml
. On the other
hand, it was found increased in ‘under stress’ normozoos-
permic (+55%; P< 0.01), oligozoospermic (+48%;
P< 0.01) and asthenozoospermic (+49%; P< 0.01)
subjects (Table 2). After treatment with M. pruriens, the
levels of lipid peroxides were reversed significantly in
normozoospermic (34%: P< 0.001), oligozoospermic
(27%; P< 0.001) and asthenozoospermic men (28%;
P< 0.001). We also observed that SOD activity in seminal
plasma of control group was 8.1 0.5 unitmg
However, this enzyme was found significantly suppressed
CON Pre Post Pre Post Pre Post
Normo Oligo Astheno
Semen volume
Sperm motility
75 Semen lique faction time
*** ***
Sperm concentrations
CON Pre Post Pre Post Pre Post
Normo Oligo Astheno
CON Pre Post Pre Post Pre Post
Normo Oligo Astheno
CON Pre Post Pre Post Pre Post
Normo Oligo Astheno
Figure 1. Semen profiles of ‘under stress’ infertile men treated with M. pruriens. CON = Control; Pre = Pre-treatment; Post = Post-treatment;
Normo = Normozoospermic; Oligo = Oligozoospermic; Astheno = Asthenozoospermic. Each bar represents mean SD. Significance:
a = P < 0.001, b = P < 0.01, c = P < 0.05; compared with controls, *** = P < 0.001; compared with pre treatment subjects, NS = Not significant.
Table 1. State anxiety score and serum cortisol levels in infertile men before and after treatment with M. pruriens
Stress parameter Non stress (Control) Psychological stress
Normozoospermic Oligozoospermic Asthenozoospermic
State anxiety score 41.0 8.0 50.1 11.8
58.6 5.6
62.2 10.4
Serum cortisol (mg/dl) (0800 h) 10.2 0.2 Pre 14.1 3.0
Pre 21.5 0.7
Pre 28.0 1.0
Post 11.6 0.4*** Post 10.4 0.5*** Post 12.6 2.0***
Serum cortisol (mg/dl) (1600 h) 5.0 0.6 Pre 10.1 1.6
Pre 13.3 4.6
Pre 16.8 1.3
Post 6.0 0.3*** Post 7.3 2.1*** Post 8.5 0.8***
Values are expressed as mean SD.
Significance: State anxiety score:
P< 0.01, as compared with the control group.
P< 0.01, as compared with the control group; ***P< 0.001, as compared with the pre treatment group.
Pre = Pre-treatment; Post = Post-treatment.
eCAM 2010;7(1) 141
in different groups of infertile men, who were under stress;
such as normozoospermic (19%; P< 0.001), oligozoos-
permic (33%; P< 0.001) and asthenozoospermic
(29%; P< 0.001). Treatment with M. pruriens increased
the activity of SOD in normozoospermic (+3%; P< 0.01),
oligozoospermic (+33%; P< 0.01) and asthenozoosper-
mic (+18%; P< 0.01) men. Similarly, catalase activity
in seminal plasma of ‘under stress’ asthenozoospermic
men was found significantly reduced (30%; P< 0.05),
as compared with healthy fertile men. Treatment with
M. pruriens enhanced the activity of the aforementioned
enzyme in all the infertile men.
Ascorbic acid levels in seminal plasma of control group
were 2.3 0.2 mg dl
. On the other hand, these levels were
found decreased in different groups of infertile men who
were under stress, i.e-normozoospermic (9%; P< 0.01),
oligozoospermic (23%; P< 0.01) and asthenozoosper-
mic (36%; P< 0.01). After treatment with M. pruriens
the levels of ascorbic acid were found increased in
normozoospermic (+11%; P< 0.5), oligozoospermic
(+33%; P< 0.01) and asthenozoospermic (+36%;
P< 0.01) men (Table 2). Similarly, GSH content in
seminal plasma of under stress normozoospermic (13%;
P> 0.05), oligozoospermic (20%; P> 0.05) and asthe-
nozoospermic (47%; P< 0.05) infertile men was found
decreased as compared with control group. Treatment with
the drug restored the levels of GSH in normozoospermic
(+19%; P< 0.05), oligozoospermic (+18%; P< 0.05)
and asthenozoospermic (+22%; P< 0.05) men.
We observed that oral administration of M. pruriens to
infertile men for 3 months not only resulted in general
improvement in sperm count and motility but it also led to
significant reduction in the level of psychological stress, as
assessed by a questionnaire and serum cortisol levels.
Male fertility and reproduction are known to be affected
by various kinds of stressful conditions, including
psychological stress (26). The autonomic nervous system
and adrenal hormones participate in stress response, which
also affects steroidogenesis and spermatogenesis (27). We
report elevated serum cortisol levels in infertile men, who
were under psychological stress. The latter causes stimula-
tion of hypothalamic-pituitary-adrenal axis (HPA) leading
to the release of the corticotropin releasing hormone
(CRF), adrenocorticotropin hormone (ACTH) and corti-
sol. Chronically increased cortisol level, as seen during
prolonged stress, may reduce the functional activity of
leuteinizing hormone—release hormone (LHRH) pulse
generator, which may lead to decrease in gonadotropin
and testosterone levels (26,28). Moreover, long-term
psychological stress may also decrease the concentration
of catchecholamines, like dopamine, noradrenalin,
5,6,dihydroxy phenyl acetic acid (DOPAC) and homo-
vanillic acid (HVA) in brain. Decrease in the activity of
dopaminergic neurons is also known to affect the fertility,
sperm count and motility. There are reports that
M. pruriens is also rich in L-DOPA, besides having several
other alkaloids and flavonoids. Therefore, our results
demonstrating reduction in psychological stress following
administration of M. pruriens may be linked to high
L-DOPA content of this herb (29).
Our results also demonstrate that lipid peroxide levels
were significantly high in the seminal plasma of subjects
who were under stress, which may be due to increased
oxidative stress. Psychological stress is known to be
associated with increased oxidant production and long-
term exposure to stress may lead to peroxidation of
polyunsaturated fatty acids of sperm membrane, resulting
in unfavorable alterations in sperm structure and func-
tion (6,7). Moreover, we also observed that in infertile
men who were under psychological stress, there were low
seminal plasma SOD and catalase activities and reduced
levels of glutathione and ascorbic acid. But there was
improvement in these enzymes and molecule levels
following treatment with M. pruriens. The improvement
in anti-oxidant content following treatment may be due
Table 2. Biochemical parameters of under stress infertile men before and after treatment with M. pruriens
Biochemical Parameter Non-stress
Psychological stress
Normozoospermic Oligozoospermic Asthenozoospermic
Pre-treatment Post-treatment Pre-treatment Post-treatment Pre-treatment Post-treatment
Lipid Peroxide (nmole MDA ml
) 2.2 0.2 3.4 0.3
2.3 0.3*** 3.3 0.3
2.4 0.4*** 3.3 0.3
2.4 0.3***
Superoxide Dismutase
(Unit mg
8.1 0.7 6.5 0.5
6.7 0.5
5.4 0.4
7.2 0.7*** 5.7 0.5
6.8 0.7***
Catalase (Unitmg
protein) 9.2 0.8 8.60.7
9.3 0.8** 8.8 0.6
9.2 0.3* 6.5 0.9
7.2 0.7***
Ascorbic acid (mgdl
) 2.3 0.2 2.0 0.2
2.3 0.2*** 1.7 0.3
2.3 0.4*** 1.5 0.3
1.2 0.3***
Glutathione (mg dl
) 1.6 0.5 1.3 0.2
1.6 0.3** 1.3 0.2
1.6 0.5** 1.3 0.3
1.6 0.3**
Fructose (mg ml
) 2.6 0.4 2.2 0.2
2.6 0.4
1.6 0.4
2.4 0.3** 1.1 0.3
1.3 0.3**
Values are expressed as mean SD.
P< 0.001,
P< 0.01,
P< 0.05 as compared with the control group.
***P< 0.001, **P< 0.01, *P< 0.05 as compared with the pre treatment group,
Not significant.
142 Mucuna pruriens in infertile men
to the reduction of oxidative stress. As stated earlier,
M. pruriens is reported to contain many bioactive
constituents, including alkaloids, coumarins, flavonoids
and alkylamines etc. which play an important role in
increasing the antioxidant capacity of treated men (15).
Furthermore, reduced stress and reactivation of antiox-
idants might have in turn lead to reduction in seminal
plasma lipid peroxide content. Our results are also in
concurrence with earlier reports that M. pruriens is a
known adaptogen and its alcoholic extract reduces lipid
peroxidation and maintains the levels of glutathione and
SOD activity (30,31).
M. pruriens seeds are rich source of L-DOPA and its
metabolites, which include epinephrine and norepineph-
rine. Therefore, an increase in dopamine level in the brain
following M. pruriens treatment may not only induce the
activation of sexual behavior but it may also increase
plasma testosterone level. It has been reported recently
that L-DOPA and its metabolite dopamine stimulate the
hypothalamus and forebrain to secrete gonadotropin-
releasing hormone (GnRH) (32). This, in turn, upregu-
lates the anterior pituitary gland to secrete follicle
stimulating hormone (FSH) and luteinizing hormone
(LH) causing increased synthesis of testosterone by
Leydig cells of the testis (33,34). Furthermore, sperma-
togenesis is controlled by the hypothalamus and anterior
pituitary working together. On the basis of the afore-
stated facts, it may be proposed that increased dopamine
level in the brain may not only optimize the release of
hormones, including testosterone, leading to increased
sexual drive and improved performance, but it may also
accomplish reduction of psychological stress. Moreover,
treatment with M. pruriens may also contribute to proper
functioning of male genital system and facilitate sperm
transport, contraction of seminal vesicles and inhibition
of lipid peroxidation of spermatozoa (35).
On the basis of results of the present study and as
gleaned by earlier reports, it may be concluded that
M. pruriens not only helps in reducing psychological
stress, but also improves semen quality as it restores
antioxidant levels and reduces lipid peroxide content.
Authors acknowledge with thanks the financial support
from Indian Council of Medical Research, New Delhi in
the form of Ad-hoc research scheme No. 5/10/8/2004-
RHN. Authors acknowledge with thanks the help and
assistance of Prof. Mahdi Hasan (Department of
Anatomy), Prof. R.K. Singh (HOD, Biochemistry) and
Dr Ramesh Chander (Department of Biochemistry).
Moreover, thanks are also due to Dr Farzana Mahdi,
Director Academics, Era’s Lucknow Medical College and
Hospital, Lucknow, for providing facilities for some of
the laboratory work. We would also like to thank
Mr M.P.S. Negi, Biometry and Statistics Division,
CDRI, Lucknow for providing assistance in statistical
analysis of data and preparation of graphs.
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Received April 3, 2007; accepted October 28, 2007
144 Mucuna pruriens in infertile men
... From the present results, it can be hypothesized that some phytochemicals probably protect the cells against sudden variation of external cues through modulation of certain transcription factors associated with expression of several enzymes and amend the initial response for resilience. Mucuna seeds were earlier reported to possess antioxidant, hypoglycemic, anti inflammatory, lipid metabolism and neuroprotective activities (Shukla et al., 2010;Ulu et al., 2018). It contains dodecanoic acid and lauric acid, which are reported for their antioxidant activity. ...
... 9,12 octadecanoic acid methyl ester in Mucuna seeds is reported to have hepatoprotective role and is also an anti-inflammatory compound (Saikarthik et al., 2017). It also possesses a number of other bioactive substances, including tryptamine, alkylamines, steroids, flavonoids, coumarins, cardenolides and metals like magnesium, copper, zinc, manganese and iron (Shukla et al., 2010). Dietary administration of Mucuna extract was also reported to improve resistance against osmolarity and salinity fluctuations and therefore speculated to have significant contribution under global climate change conditions (Moniruzzaman et al., 2018). ...
Increases in ambient temperature affect the biochemical status of fish, and dietary supplementation with bioactive phytoconstituents may promote resilience against environmental stress. This study evaluated the impact of three plant extracts on the biochemical status of a cold stream fish Botia rostrata (Günther, 1868) under high temperatures. After 1 month dietary supplementation separately with Mucuna pruriens methanol extract (0.25g/kg feed), Tribulus terrestris ethanol extract (0.5g/kg feed) and Basella alba ethanol extract (1.0g/kg feed), juvenile fish (Wt. 4.3 ± 0.5g) were exposed to different sublethal heat stress [28 ± 0.5 °C (T1), 32 ± 0.5 °C (T2), 36 ± 0.5 °C (T3)]. Control fish were fed a diet without any plant extract and maintained at 24 ± 0.5 °C. Serum and muscle tissues were collected to measure different biochemical parameters, muscle metabolic enzymes and molecular chaperons before and after heat stress. Before stress, the group fed the Mucuna diet showed significant (P < 0.05) increases in serum glucose [+10.92%], protein [+18.93%], muscle heat shock protein (HSP) 90 [+8.6%] compared to the control group. No significant change (P > 0.05) of stress parameters was observed between control, Tribulus and Basella fed fish. The control group exposed to T3 showed significant differences (P < 0.05) in protein [-26.19%], lactate dehydrogenase [+93.69%], fructose 1,6 bisphosphate [-35.19%], phosphorylase ‘a’ [+35.72%], HSP60 [+69.54%], HSP70 [+84.85%], HSP90 [+92.07%], heat shock factor (HSF) 1 [+88.48%] suggesting susceptibility of Botia to this temperature. Among the three plant extracts, Mucuna methanol extract was effective to enhance resistance against temperature-induced biochemical alterations. After exposure to T3, only the fish fed Mucuna diet showed no mortality. Fish fed Mucuna diet exposed to 36 ± 0.5 °C showed significantly higher (P < 0.05) glucose [+42.82%], protein [+11.98%], citrate synthase [+59.81%], phosphorylase ‘a’ [+14.96%], Glucose 6 phosphate dehydrogenase [+60.87%], HSP60 [+34.13%], HSP70 [+41.42%], HSP90 [+65.91%], HSF1 [+61.32%] compared to those in Mucuna fed fish maintained at 24 ± 0.5 °C. These results highlight temperature-induced biochemical alterations in Botia and point towards the potential use of Mucuna in overcoming such adverse high thermal stress.
... 1e3 In men, clinical studies have reported that chronic stress significantly increased serum cortisol level and reduced the sexual performances, androgen levels, and semen quality parameters. 2,4,5 To explore more mechanism of male infertility caused by stress, the chronic unpredictable mild stress (CUMS) model has been widely used in experimental animals by randomly exposure to many unpredictable stressors to mimic the physiological symptoms of human depression. 6,7 Previous studies reported that CUMS could impair male sexual behaviors and damaged the seminiferous and epididymal tissues in rodents. ...
... 17e19 Moreover, M. pruriens seed has been proven to improve semen quality and antioxidant enzymes in seminal plasma of infertile men. 5 In rodents, M. pruriens could reduce the damages of sperm DNA and testicular mitochondria. 20,21 Mucuna pruriens (L.) DC. var. ...
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Background and aims Chronic stress is a major common cause of male infertility. Many species of velvet beans are shown to be rich in l-DOPA. In Thai folklore medicine, seeds of Mucuna pruriens (L.) DC. var. pruriens (Thai Mhamui or T-MP) have been used for treating erectile dysfunction. This study aimed to determine l-DOPA levels in T-MP seed extract and investigate its preventive on sexual behaviors and reproductive parameter damages including essential proteins in chronic unpredictable mild stress (CUMS) mice. Experimental procedure Mice were divided into 4 groups: (I) control, (II) CUMS, (III) T-MP300 + CUMS, and (IV) T-MP600 + CUMS. Groups I and II received DW while groups III and IV were pretreated with the seed extracts (300 and 600 mg/kg BW) for 14 consecutive days before co-treatment with a randomly different CUMS/day (from 12 mild stressors) for 43 days. Results and conclusion T-MP seed extract contained l-DOPA approximately 10% of total dried weight. A dose of 600 mg/kg improved sexual performances and degenerative seminiferous epithelium in CUMS mice. Sperm qualities and testosterone level were elevated while corticosterone was decreased in co-treatment groups. T-MP-CUMS cotreated groups also improved expressions of AKAP4, AR, and TyrPho proteins in testis, epididymis, and sperm. T-MP increased StAR and CYP11A1 expressions in testis. It also suppressed testicular apoptosis via decreased expressions of Hsp70, caspases 3, and 9. T-MP seeds containing l-DOPA could improve sexual behaviors and essential reproductive proteins caused by CUMS. Section Natural Products; Taxonomy (classification by evise) Traditional Herbal Medicine; Animal Model; Histopathology.
... Infertile subjects were administered with M. pruriens seed powder (5g per day) orally. According to the results, it may be concluded that M. pruriens not only reactivates the antioxidant defense system of infertile men but it also helps in the management of stress and improves sperm concentration and motility 17 . As proven by Sharma, T. et al the black seeds of M. pruriens comparatively give more percentage improve on all subjective parameters i.e. ...
... There are also reports that the methanol extract of M. pruriens seeds has strong antioxidant activity, because it inhibits 1,1diphenyl-2-picryl-hydrazyl and hydroxyl radical, and that it also has nitric oxide and superoxide anion scavenging and hydrogen peroxide decomposing and reducing power 14,26 . It also proved that M. pruriens not only reactivates the anti-oxidant defense system of infertile men but it also helps in the management of stress and improves semen quality 17 . Furthermore, M. pruriens seeds contain magnesium and zinc (8.74-19.38 mg and 0.25-0.54 ...
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In Ayurveda Mucuna pruriens (M. pruriens) is considered as a medicine for male subfertility. There are in vitro, in vivo and literature reviews to demonstrate beneficial medicinal effects of M. pruriens in worldwide. This study aims to systematically review the scientific literature and provide a comprehensive summary on the effect of M. pruriens on sperm parameters with idiopathic infertility. A comprehensive systematic review was conducted in the following databases: PubMed, Google scholar and Cochrane library for research articles published in between 2000 and 2020. The keywords that were used in combination to search for articles included, "Mucuna pruriens","idio-pathic infertility" and "sperm parameters". The literature search done by using keywords and identified following number of articles in the respective databases; PubMed (n=13), Google scholar (n= 116), and Cochrane library (n=4). After removing duplications, the total number of articles included in the present review is 124. 114 studies were omitted due to exclusion criteria and use of combine M. pruriens with other medicinal herbs, due to the impossibility of determining the net effect of M. pruriens. Included articles were screened in the final stage by reading the full article. There were 10 different studies evaluating the in vitro effect of M. pruriens on sperm parameters with idiopathic infertility. The results of this study showed that consumption of M. pruriens significantly improves
... Sexual potency and behavior, libido, sperm parameters, and endocrine levels all improved significantly when M. pruriens seed extract was used (Suresh and Prakash 2012). Lipids, triglycerides, cholesterol, phospholipids vitamin A, C, and E levels have all been improved by M. pruriens and restored the levels of SOD, catalase, GSH, and ascorbic acid in seminal plasma after fructose nullified oxidative stress-induced lipid peroxidation in seminal vesicles Shukla et al. 2010). The Mucuna pruriens increases testosterone, LH, dopamine, adrenaline, and noradrenaline levels whilst declining FSH and PRL levels in infertile men, according to neuroendocrine studies. ...
Fertility of male animals is influenced by various factors such as neurohormonal imbalances, reproductive organ tissue changes, seminal attributes, libido and sexual behavior. The global decline in male reproductive health is a big worry, and modern therapeutic options to prevent male infertility are costly, less accessible, require long-term treatment, and have a variety of adverse effects. Herbal remedies, on the other hand, are better suited to providing more comprehensive approaches to improve the male reproductive health. There is a particular set of herbs known as vajikarana or aphrodisiacs in Ayurvedic pharmacology that nourishes and stimulates the sexual tissues. This review focuses on the Ayurvedic approach to improving male reproductive health, referring to some of the most important scientifically tested herbs that have been found to boost male fertility by having stimulating or nourishing effects on the male reproductive organs.
... The deficiency of vitamin C may produce oxidative damage induced by reactive oxygen species (ROS); an increase in ROS was observed in the semen of infertile men [74]. In a human trial, a decrease in vitamin C levels was associated with an increase in seminal plasma lipid peroxidation [75]; it may also lead to abnormal sperm parameters [76]. Low concentration of vitamin C produced marked degenerative changes in the testes, epididymis, and vas deferens of scorbutic guinea pigs [77], also produced degeneration of the spermatogenic epithelium, steroidogenesis, and decline in plasma testosterone levels [78]. ...
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Vitamin C is associated with history of the cause of the ancient hemorrhagic disease scurvy. Vitamin C is an essential nutrient with important antioxidant properties. It is required by the body for normal physiological function. The body cannot synthesize vitamin C, it is present in nature through foods and other natural sources and it exists as a nutritional food supplement. The antioxidant activity of vitamin C protects the body from free radical damage. Vitamin C is essential for the development and maintenance of connective tissues. It is used as therapeutic agent in many diseases and disorders. Vitamin C plays an important role in several metabolic functions, as the conversion of the amino acid, tryptophan, to the neurotransmitter, serotonin, and the conversion of cholesterol to bile acids. Vitamin C supplementation resulted in a significant increase in vitamin C levels in populations; its high intake is associated with positive effects on cardiovascular risk factors. Vitamin C protects the immune system, reduces the severity of allergic reactions and helps to fight infections. It has an important role in bone formation, wound healing and the maintenance of healthy gums. There is profound beneficial effect of vitamin C in respect to human diseases as cancer, atherosclerosis, diabetes, neurodegenerative disease and many metal toxicities. Several vitamin C analogs have been produced as anticancer and antioxidant activity. Vitamin C is useful if it is used as adjuvant therapy for several chronic diseases. Thus, this review summarizes the importance of vitamin C in the body’s physiology and biochemistry, in addition, the different mechanisms that vitamin C is implicated to treat different acute and chronic diseases. Future exploration should pay attention to chronic disease management by vitamin C.
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Ayurveda is an ancient science of life that mainly deals with 'Ayu' where the Ayu is combination of Sharira, Indriya, Satva & Atma. Ayurveda is Upaveda of Atharveda. It is the science of life based mainly upon Tridosha Siddhant, Ayurveda has been divided in to eight branches, out of which Shalakya Tantra is one of them. Shalakya Tantra is the science of diseases and their management specific to Urdhvangas. शाल􀃈यम ् नामो􀃚वज
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Purpose: Testosterone replacement and associated pharmacologic agents are effective strategies to treat male hypogonadism; however, nutraceutical agents and lifestyle modification approaches have gained medical interest. The purpose of this scoping review is to highlight the evidence (or lack thereof) of nutraceuticals and lifestyle modification approaches in the management of testosterone levels and sperm parameters. Methods: A scoping review of nonpharmacologic interventions (supplements, herbal medicines, diets, sleep, and exercise) with the potential to improve male health was undertaken to elucidate changes in testosterone levels and sperm parameters in men with hypogonadism or infertility compared with healthy patients. Findings: A multitude of nutraceuticals and functional nutrients are purported to stimulate testosterone production; however, only a select few have had promising results, such as zinc, vitamin D (in case of hypovitaminosis D), l-arginine, mucuna, and ashwagandha, based on well-controlled randomized clinical trials of men with low testosterone levels and related problems. Except for l-arginine, these natural agents, as well as tribulus and ω3 fatty acids, can improve some degree of sperm parameters in infertile men. Before implementing these nutraceutical agents, adequate sleep, exercise, and weight loss in patients with obesity are imperative. The effects of nonpharmacologic interventions on testosterone levels are modest and hence do not directly translate into clinical benefits. Correspondingly, androgen receptor content, but not endogenous androgens, has been regarded as the principal factor in muscle hypertrophy. Implications: A limited number of supplements and herbal medicines can be considered as adjunctive approaches in the management of testosterone levels and sperm parameters, primarily in men with low testosterone levels and infertility, whereas most nonpharmacologic supplements appear to lack evidence. Although proper physical exercise, sleep, and diet are indisputable approaches because of the general benefits to health, the use of nutraceuticals, if considered, must be personalized by physicians and/or registered dietitians.
Infertility is a significant cause of anxiety, depression, and social stigma among couples and families. In such cases, male reproductive factors contribute widely to the extent of 20-70%. Male infertility is a multifactorial disease with several complications contributing to its diagnosis. Although its management encompasses both modern and traditional medicine arenas, the first line of treatment, adopted by most males, focuses on the reasonably successful medicinal plant-based conventional therapies. Phyto-therapeutics, which relies on active ingredients from traditionally known herbs, influences sexual behavior and male fertility factors. The potency of these phyto-actives depends on their preparation methods and forms of consumption, including decoctions, extracts, semi-purified compounds, etc., as inferred from in vitro and in vivo (laboratory animal models and human) studies. The mechanisms of action therein involve the testosterone pathway for stimulation of spermatogenesis, reduction of oxidative stress, inhibition of inflammation, activation of signaling pathways in the testes [extracellular-regulated kinase (ERK)/protein kinase B(PKB)/transformation of growth factor-beta 1(TGF-β1)/nuclear factor kappa-light-chain-enhancer of activated B cells NF-kB signaling pathways] and mediation of sexual behavior. This review critically focuses on the medicinal plants and their potent actives, along with the biochemical and molecular mechanisms that modulate vital pathways associated with the successful management of male infertility. Such intrinsic knowledge will significantly further studies on medicinal plants that improve male reproductive health.
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Abstract Ethnopharmacological relevance Thai Mucuna pruriens (L.) DC. var. pruriens (T-MP) has been traditionally used in treating depressive disorders, dysuria and enhancing male sexual desire. Although T-MP seed is demonstrated to have antioxidant capacity, its aphrodisiac and protective tissue damage properties have never been documented. Recently, ethanol (Eth) is known to cause sexual behavior dysfunction and damage reproductive system. This study aimed to investigate the protective effects of T-MP seed extract on sexual behavior dysfunction and reproductive damages in male rats admisted with Eth. Materials and methods T-MP possessing antioxidant activity was determined for L-DOPA content using NMR analysis. Thirty-six male rats were divided into four groups (9 animals/group). Control rats received DW and the ethanol (Eth) group was given with Eth (3 g/kgBW; 40%v/v). In preventive groups (T-MP150 + Eth and MP300 + Eth groups), animals were treated with T-MP extract at a dose of 150 and 300 mg/kgBW before Eth administration for consecutive 56 days. Sexual behaviors including mounting frequency (MF), intromission frequency (IF), mounting latency (ML), intromission latency (IL), ejaculation latency (EL), post-ejaculatory interval (PEI), and ejaculation frequency (EF) were evaluated. Epididymal sperm quality and daily sperm production (DSP) were examined. Testicular histology was observed using Masson's trichrome staining. The malondialdehyde (MDA) levels and expressions of androgen receptor (AR), heat shock protein 70 (HSP70), steroidogenic acute regulatory (StAR), and tyrosine-phosphorylated (TyrPho) proteins in testis were also determined. Results T-MP extract contained L-DOPA and improved sexual behaviors including increased MF and IF and decreased ML and IL in Eth treated rats. Significantly, sperm quality, DSP, and testicular histopathology observed in Eth group were improved after T-MP treatment. T-MP also decreased the testicular MDA levels. Additionally, T-MP could correct testicular functional proteins of AR and StAR except HSP70 expression in Eth group. Expressions of TyrPho proteins in testicular and sperm lysates were improved in co-administered groups. Conclusions T-MP seed extract possessing L-DOPA could enhance the sexual behaviors and protect reproductive damages via improvement of testicular functional proteins.
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An enzyme which catalyzes the dismutation of superoxide radicals (O2·⁻ + O2·⁻ + 2H⁺ → O2 + H2O2) has been purified by a simple procedure from bovine erythrocytes. This enzyme, called superoxide dismutase, contains 2 eq of copper per mole of enzyme. The copper may be reversibly removed, and it is required for activity. Superoxide dismutase has been shown to be identical with the previously described copper-containing erythrocuprein (human) and hemocuprein (bovine). Stable solutions of the superoxide radical were generated by the electrolytic reduction of O2 in an aprotic solvent, dimethylformamide. Slow infusion of such solutions into buffered aqueous media permitted the demonstration that O2·⁻ can reduce ferricytochrome c and tetranitromethane, and that superoxide dismutase, by competing for the superoxide radicals, can markedly inhibit these reactions. Superoxide dismutase was used to show that the oxidation of epinephrine to adrenochrome by milk xanthine oxidase is mediated by the superoxide radical. An assay of several tissues indicates that superoxide dismutase is widely distributed within mammalian organisms.
We undertook this study to investigate lipid peroxide levels and the activities of two enzymes, superoxide dismutase and catalase, in fertile (n = 66) and normozoospermic infertile, origozoospermic, asthenozoospermic and azoospermic males (n = 92). The results show significantly high seminal plasma lipid peroxide levels in normozoospermic infertile and asthenozoospermic men. Moreover, we found significantly reduced activity of the enzyme superoxide dismutase in all groups, except for those having oligozoospermia. Catalase activity was significantly lower in all infertile males. Elevated lipid peroxide levels along with depressed antioxidant enzyme activities highlight the role of free radicals in male infertility.
Mucuna pruriens seed powder when administered in a dose of 75 mg/kg body weight daily as an aqueous suspension, increased the sexual activity of male albino rats considerably. The different components of copulatory behaviour vis. mount frequency, mount latency, intromission frequency and intromission latency were found to be influenced by the test drug. L-dopa, one of the constituents of the title plant was also reported to arouse sexual desire in patients suffering from Parkinson's disease. At 100 mg/kg body weight, though L-dopa was found to arouse sexual activity, the low content of this constituent (0.69%), could not account for the activity shown by the seed powder in the dose employed.
Testosterone is the main androgen in the male, mainly synthesized by the Leydig cells in the interstitial tissues of the testes. It is a pro-hormone for biologically active 5-areduced androgen and estrogens and is produced by the action of luiteinizing hormone (LH) through cyclic adenosine monophosphate (cAMP) pathway. This classical pathway of steroidogenesis is a major but not sole player in testosterone synthesis in the Leydig cells. Other intracellular signaling systems also play important role in Leydig cells via which several endogenous bio-chemicals have been found to actively participate either in stimulation or inhibition of testosterone production. One of the factors that have been poorly investigated for their autocrine and paracrine action in the testosterone production by the Ledyig cells are the neurotransmitters. In this experiment we investigate some putative neurotransmitters (NTs) namely dopamine (DA), 3, 4-dihydrophenylacetic acid (DOPAC), epinephrine (E), norepinephrine (NE), 5-hydroxytryptamine (5-HT), homovanillic acid (HVA), 3-methoxytyramine (3-MTA), 5-hydrooxyindoleacetic acid (5-HIAA) for their direct effect on the testosterone production by isolated interstitial cells from mice. The finding reveal that DA, 5-HT at lower doses can acutely stimulate while DOPAC, E, 3-MTA inhibits testosterone production significantly over the basal level (p<0.05). At longer duration of incubation the stimulation and inhibition of either NTs is not significantly different then the basal except for high dose of mg/ml 5-HT which showed significant inhibition both at 4 and 24 hours of incubation over the basal level. The exact mechanism how some of these putative NTs can acutely stimulate testosterone production while others can inhibit is a matter of further investigation. It is postulated that enzymatic metabolization of 5-HT to 5-HIAA and DA to 3-MTA is partly responsible for the significant inhibition of testosterone production acutely and at longer duration of incubation as seen with higher dose. Further investigation is necessary before we can come to any conclusive decision regarding their exact mechanism but NT=s local role in testosterone production from Leydig cells is an crucial clue to unfold the unique mystery of testosterone synthesis.
The effects of different drugs influencing brain neurotransmitter contents have been tested on the pituitary-testicular function in male rats. L-dopa (200 mg/kg body weight, i.p.) increased the dopamine and noradrenaline contents of the hypothalamus, amygdala, striatum and mesencephalon, but it was ineffective as regards the 5-hydroxytryptamine contents of the same brain areas, and increased the plasma testosterone level. alpha-Methyl-p-tyrosine (250 mg/kg b.w., i.p.) decreased the dopamine and noradrenaline contents of these brain areas, but it was ineffective to 5-hydroxytryptamine, and decreased the plasma testosterone level. Diethyldithiocarbamate (400 mg/kg b.w., i.p. twice a day) increased the dopamine levels in the hypothalamus, amygdala, striatum and mesencephalon, decreased the noradrenaline contents in the same brain regions but had no effect on the 5-hydroxytryptamine contents of these brain areas or on the testosterone level in the peripheral blood. p-Chlorophenylalanine (300 mg/kg b.w., i.p.) decreased the 5-hydroxytryptamine contents of the different brain areas, while it had no effect on the dopamine and noradrenaline levels or on the plasma testosterone level. 5-Hydroxytryptophan (200 mg/kg b.w., i.p.) increased the 5-hydroxytryptamine contents of all brain areas studied, but was without effect on the dopamine and noradrenaline contents or the plasma testosterone level. The data suggest that both dopamine and noradrenaline may be involved in the regulation of the pituitary-testicular function, and the ratio of the two transmitters might be more important that their actual levels in definite brain areas.