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Bangladesh Journal of Medical Science Vol.10 No.2 Apr’11 

1. *OE Yama,  
2. FIO Duru,  
3. AA Oremosu,  
4. CC Noronha 
Department of Anatomy, College of Medicine, University of Lagos, Idi-Araba, Lagos, Nigeria.  
*CorrespondS to: Dr. Yama Oshiozokhai Eboetse, Department of Anatomy, College of medicine of the 
University of Lagos, P.M.B. 12003, Lagos, Nigeria, Email: dro_yama@yahoo.com. 

Original article:  

Testicular oxidative stress in Sprague-Dawley rats treated with bitter melon 
(Momordica charantia): the effect of antioxidant supplementation 

 
OE Yama1, FIO Duru2, AA Oremosu3, CC Noronha4 

 
Abstract 

Objective: An important mediator of testicular injury is oxidative stress; the implicating pathway 
has been pointed at a free radical mechanism by researchers. This article, investigates the effect of 
bitter melon (Momordica charantia) (MC) seed extract and antioxidant supplementation in the 
testes of Sprague-Dawley (S-D) rat. Methodology: Ninety male S-D rats, weighing between 110-
214 g, were assigned randomly into six main Groups A to F. Group A was administered 50 mg/100 
g of MC extract orally, between 6 to 16 weeks. Group B were pre-treated with ascorbic acid (AA) 
0.01mg/kg, three days/week, α-tocopherol (AT) 20 mg/kg, five days/week and both test solutions 
(TS) i.e. AA and AT; 0.01 and 20 mg/kg, three and five days/week for 8 weeks. This was followed 
by administration of the extract at dose and duration as in A. Group C received the extract for 8 
weeks and afterwards post-treated for another 8 weeks with AA, AT and both TS (as above). 
Group D in addition to the extract administration were treated with AA, AT and both TS in dose 
and duration similar to B above. Group E had AA, AT and both TS alone for 8 weeks. Group F 
served as the control subjects. The animals testicular tissues were processed for malondialdehyde 
(MDA) and AA concentrations. Serum testosterone (TT) assay was done from left ventricular 
blood. Results: The extract administered for 6, 8 and 16 weeks produced significantly (p < 0.05) 
increased testicular MDA (1.74 ± 1.15, 1.84 ± 0.38 and 2.38 ± 0.40) compared to control (0.38 ± 
0.02, 0.38 ± 0.03 and 0.35 ± 0.02) and decreased AA (0.01± 0.02, 0.01± 0.01 and 0.00± 0.01) 
compared to control (0.15 ± 0.02, 0.12 ± 0.02 and 0.13 ± 0.02). There was also an associated 
significant decrease (p < 0.05) in peripheral TT levels compared to control. The extract produced 
responses that showed no prophylactic rather modulatory effect with TS. Conclusion: These 
findings suggest that the extract resulted in changes in the testicular oxidative status. This may 
play a role in testicular dysfunction that may compromise fertility.  
 
Key words: Momordica charantia, malondialdehyde, ascorbic acid, testosterone. 

Introduction 

Over seven decades ago, α-tocopherol (AT) 
was recognized as a powerful lipophilic 
antioxidant that is absolutely vital for the 
maintenance of mammalian spermatogenesis 
[1]. Ascorbic acid (AA) contributes to the 
support of spermatogenesis at least in part 
through its capacity to reduce AT and maintain 
this antioxidant in an active state [2]. Free 
oxygen radicals are known to possess ability to 
react with cellular macromolecules such as 
nucleic acids, lipids, proteins and carbohydrates 
to produce a destructive effect [3]. For example 

oxygen radicals have a destructive effect on 
lipids (lipid peroxidation) of all membranes. The 
end product of this phenomenon is called 
malondialdehyde (MDA). It is a reliable and 
generally accepted indicator of lipid 
peroxidation [3]. The levels can increase to the 
extent that it cumulates into a situation known 
as oxidative stress [3, 4, 5]. Free oxygen radicals 
or reactive oxygen species (ROS) such as 
superoxide anions, hydrogen peroxide, and the 
hydroxyl ion are molecules that contain an 
oxygen atom. A free radical is any chemical 



Yama OE, Duru FIO, Oremosu AA, Noronha CC 

105 

species capable of independent existence and 
contains one or more unpaired electrons. They 
are highly reactive due to the presence of 
unpaired valence shell electrons. The cellular 
structures of membranes are prevented from 
the damaging effect of ROS by systems that 
scavenge the free radicals from the cellular 
environment [4].  
 
There is normally an intricate balance between 
these amount of free radicals generated and 
scavenged by a cell with damage occurring 
when the equilibrium is disturbed [6]. Thus 
when there is increased production of ROS, 
cellular structures are vulnerable to the effects 
of oxidative stress [4]. Spermatozoa are rich in 
polyunsaturated fatty acids and this makes 
them susceptible to attack by ROS or 
membrane lipid peroxide ions. The equilibrium 
between the amount of ROS produced and 
scavenged is related to the stability and 
damage of the gamete cell[4]. Free radicals are 
implicated to have detrimental effects on 
sperm functions, which depend on its nature 
and concentration [7]. Antioxidants are 
substances that inhibit the destructive effects of 
oxidation by the ROS in the body [8]. 
Numerous antioxidants include ascorbic acid, 
alpha tocopherol, beta carotene and melatonin. 
Other known antioxidants include enzymes 
such as superoxide dismutase catalase and 
gluthatione peroxidase are credited to ROS 
detoxification [9]. The antioxidant properties of 
Momordica charantia has been pointed out 
previously by researchers [10] but its pro-
oxidant effect on the testes is yet to be 
described. Research has proven that at a high 
dose antioxidant could act as pro-oxidant 
releasing free radicals [11]. This present study 
was thus designed to determine the possible 
role of oxidative stress of Momordica 
charantia on the testes as a mode of 
contraception in male S-D rats previously 
described.  
 
Materials and methods  

Test solutions 
The Test solutions (TS) used were the 
antioxidants α-tocopherol (AT) and ascorbic 
acid (AA) at doses 20 mg/kg [12] and 0.01 

mg/kg [13] respectively.The doses were 
calculated based on the animal’s individual 
weekly body weights and aliquots 
approximated to the nearest numeral 
administered. It was done using insulin syringe 
(100 IU equivalent to 1ml) via intramuscular 
(i.m.) route.  
 

Collection and identification 
The ripe fruits of MC harvested in June were 
purchased from the local market in Lagos. It 
was authenticated by Professor J. 
Olowokudejo, a taxonomist in the Department 
of Botany, University of Lagos, where the 
voucher specimen was deposited (ascension 
number FHI 108422). 
 

Preparation of seed extract 
The seeds were dried in an oven (temperature 
of between 30-40oC) for a week. The dried 
seeds were weighed and Soxhlet extraction 
done using alcohol and water as solvents at the 
Pharmacognosy department of College of 
Medicine, University of Lagos. The percentage 
yield was 23.0% w/w, from which a dose of 50 
mg/100 g of body weight was calculated and 
administered orally.  
 

Sources and maintenance of Rats  
Ninety male S-D rats 6-8 weeks old were used 
in this study. The rats were procured from the 
Laboratory Animal Centre of the College of 
Medicine of the University of Lagos and were 
authenticated by a taxonomist [14] in the 
Department of Zoology of the University of 
Lagos. They were kept in plastic cages in the 
Animal Room of the Department of Anatomy 
and allowed to acclimatize for two weeks 
under standard laboratory conditions of 
temperature 27-30oC. Lighting was by natural 
daylight such that the rats were exposed to 
approximately 12:12 light–dark cycle. They 
were fed with commercially available rat chow 
(Livestock feeds Plc, Ikeja, Lagos, Nigeria) 
and had unrestricted access to water.  
 

Experimental protocol 
The animals were randomly allocated into 6 
main groups A to F. Which were further sub-
divided into 3 sub-groups (A1 to A3; B1 to B3; 
C1 to C3; D1 to D3; E1 to E3 and F1 to F3) of 5 
rats. Subgroups A1 to A3 indicate different 



Testicular oxidative stress in Sprague-Dawley rats administered Momordica charantia 

106 

treatment durations of 6, 8 and 16 weeks 
administered a single oral dose of 50 mg/100 g 
of MC extract. Group B pre-treated with TS 
(BI: 0.01 mg/kg of AA, three times a week 
Mondays, Wednesdays and Fridays; B2: 20 
mg/kg of AT, five days a week Mondays, 
Tuesdays, Wednesdays, Thursdays and Fridays 
while B3: both TS) for 8 weeks and then fed 
the extract at dose and duration as in A. 
Subgroups C1 to C3 received the extract for 8 
weeks (as in Group A) and afterwards post-
treated for another 8 weeks with AA, AT and 
both TS (as in Group B). Group D comprise 
rats receiving the extract and TS 
simultaneously for a duration of 8 weeks. 
Therefore in addition to the extract D1 to D3 
were treated with AA, AT and both TS in dose 
and duration similar to B above. Subgroups E1 
to E3 had AA, AT and both TS. Finally, Group 
F animals were used to compare events in the 
other groups was administered distilled water 
throughout the experiment.  
 
At the end of the different experimental 
durations, the animals were sacrificed. The 
testes were assessed for malondialdehyde and 
ascorbic acid concentration, cauda epididymal 
fluids were processed for sperm count and 
motility. Testicular morphometry (weight and 
volume) was also assessed. The protocol was 
approved by ethical committee of the institute.  
 
Testicular malondialdehyde concentration 
Testicular tissue samples weighing 0.25 g were 
homogenized in 2.5 ml of 0.15 M potassium 
chloride. The supernatant from the homogenate 
was collected and stored at 200C. The MDA 
levels were determined as described by Buege 
and Aust (1978) [15]. A 2ml aliquot of 0.375% 
Trichloroacetic acid- Thiobarbituric acid- 
Hydrochloric acid (TCA-TBA-HCL) was 
added to 1.0ml of the supernatant of testicular 
tissue homogenate. This was mixed vigorously 
and heated for 15minutes in a water bath at 80-
900C. The sample was cooled in ice cold water 
again for 15 minutes at 1500 g and the tubes 
were placed in the photometer and absorbance 
taken at 535nm against the reagent blank. 
Concenration was calculated using the molar 
absorptivity of malondialdehyde which is 1.56 
x 105 M-1cm-1.  

Testicular ascorbic acid concentration  
Testicular AA concentrations were determined 
as by the Association of Official Analytical 
Chemist (1990) [16]. This method is based on 
the oxidation of ascorbic acid to dehydro 
ascorbic acid, which when heated with 
dinitrophenylhydrazine forms a coloured 
complex with absorption maxima at 520 nm. 
Briefly, 0.5 g testicular tissue sample was 
homogenized in 12.5 ml of 0.5% oxalic acid 
for 10 minutes. The homogenate was 
centrifuged at 1000 g and the supernatant 
collected. 1.5 ml of 4% trichloroacetate and 1.0 
ml of 2, 4-dinitrophenylhydrazine were added 
to 0.5 ml of supernatant in test tubes. The tubes 
were then incubated at 50oC for 1 hour. With 
the tubes in ice-bath, 1.25 ml of 85% sulphuric 
acid was added drop-wise with mixing after 
each drop. The tubes were removed from the 
ice-bath and left at room temperature for 30 
minutes. The absorbance was then read at 520 
nm after setting the spectrophotometer to zero 
with the blank. The concentration of ascorbic 
acid was calculated using the formula: 
Concentration of ascorbic acid = Abs(test) X 
Con(std)/Abs(std) Where, Abs(test) is the absorbance 
for the sample, Con(std), concentration of 
standard ascorbic acid and Abs(std), absorbance 
of standard ascorbic acid (0.086), derived from 
Beer-Lambert law (Absorbance proportion to 
Concentration) (Association of Official 
Analytical Chemists, 1990).  
 
Testosterone assay 
Serum TT was assayed from blood obtained 
from a left ventricular puncture. The samples 
were spun at 3000 g for 10 min in an angle 
head centrifuge at 25 oC. The samples were 
assayed in batches from a standardized curve 
using the Enzyme Linked Immunosorbent 
Assay (ELISA) method [17]. The microwell kits 
used were from Syntro Bioresearch Inc., 
California USA. Using 10 µl of the standard, 
the samples and control were dispensed into 
coated wells. 100 µl TT conjugate reagent was 
added and then 50 µl of anti-TT reagent. The 
contents of the microwell were thoroughly 
mixed and then incubated for 90 minutes at 
room temperature. The mixture was washed in 
distilled water and further incubated for 20 
minutes. The reaction was stopped with 100 µl 



Yama OE, Duru FIO, Oremosu AA, Noronha CC 

107 

of 1N hydrochloric acid. Absorbance was 
measured with an automatic spectrophotometer 
at 450 nm. A standard curve was obtained by 

plotting the concentration of the standard 
versus the absorbance and TT concentration 
was determined from the standard curve. 

 
Table I: Malondialdehyde and Ascorbic acid concentration in experimental and control Sprague-Dawley rats 

Groups  
(n = 90) 

 Testicular MDA (nmol/g 
of testis x 10-7)  

Testicular AA (mg/100 
m3 of testis) 

A1 
A2 
A3 

Wk 6 
Wk 8 
Wk 16 

1.74 ± 1.15b 
1.84 ± 0.38b 
2.38 ± 0.40b 

0.01± 0.02b 
0.01± 0.01b 
0.00± 0.01b 

B1 
B2 
B3 

AA8wks – MC8wks 
AT8wks – MC8wks 
AA8wks,AT8wks – MC8wks 

2.12 ± 0.08b 
2.35 ± 0.80b 
1.73 ± 0.34b 

0.02 ± 0.02b 
0.04 ± 0.06b 
0.02 ± 0.02b 

C1 
C2 
C3 

MC8wks – AA8wks 
MC8wks – AT8wks 
MC8wks – AA8wks,AT8wks 

0.31 ± 0.08 
0.69 ± 0.45 
0.45 ± 0.20 

0.10 ± 0.02 
0.08 ± 0.04 
0.12 ± 0.02 

D1 
D2 
D3 

MC8wks + AA8wks 
MC8wks + AT8wks 
MC8wks+ AA8wks,AT8wks 

0.37 ± 0.05 
0.59 ± 0.22 

0.30 ± 0.009 

0.19 ± 0.02 
0.16 ± 0.02 
0.21 ± 0.09 

E1 
E2

 

E3 

AA8wks  
AT8wks  
AA8wks,AT8wks 

0.34 ± 0.08 
0.39 ± 0.18 
0.36 ± 0.12 

0.14 ± 0.05 
0.12 ± 0.05 
0.11 ± 0.06 

F1 
F2 
F3 

Distilled water6wks 
Distilled water8wks 
Distilled water16wks          

0.38 ± 0.02 
0.32 ± 0.03 
0.35 ± 0.02 

0.15 ± 0.02 
0.12 ± 0.02 
0.13 ± 0.02 

Values expressed as Mean ± standard deviation; bp < 0.05; Distilled water given for 6 to 8 weeks; 
MDA: Malondialdehyde; MC8wks: 50 mg/100 g of Momordica charantia extract fed for 8wks; 
AA8wks: 0.01 of ascorbic acid treated for 8wk; AT8wks: 20 mg/kg of α-tocopherol treated for 8wk; 
AA8wks, AT8wks: α-tocopherol & ascorbic acid at doses 0.01 and 20 mg/kg administered 
simultaneously for 8wk; wk: weeks 
 
Statistical analysis 
Results were expressed as mean±SD. Analysis 
was carried out using analysis of variance 
(ANOVA) with Scheffe’s post hoc test. The 
level of significance was considered at p < 
0.05. 
 
Results 

Testicular malondialdehyde levels 
There was a marked duration dependent 
statistically significantly (p < 0.05) increase in 
testicular MDA concentration compared to 
control (Table-I). The mean values for animals 
fed distilled water and MC extract for 6, 8 and 
16 weeks were 0.38 ± 0.02, 0.32 ± 0.03, 0.35 ± 
0.02 vs1.74 ± 1.15, 1.84 ± 0.38, 2.38 ± 0.40 
respectively. This increase is also same for those 
pre-treated with AA, AT, TS for 8 weeks and 

later post-treated with the extract for another 8 
weeks, mean values were 2.12 ± 0.08, 2.35 ± 
0.80, 1.73 ± 0.34. Animals fed the extract for 8 
weeks followed by treatment with AA, AT and 
both TS for 8 week were 0.31 ± 0.08, 0.69 ± 
0.45, 0.45 ± 0.20 respectively, when compared 
to control (0.38 ± 0.02, 0.32± 0.03, 0.35 ± 0.02) 
showed no significant difference (p < 0.05; 
Table-I). The mean MDA for animals post 
treated with AT was observed to be slightly 
higher values were not statistically significant. 
Also showing no significant difference from 
control (0.38 ± 0.02) were the groups in which 
the extract was administered concurrently 
with AA, AT, TS for 8 weeks viz 0.37 ± 0.05, 
0.59 ± 0.22, 0.30 ± 0.01 (Table-I). Finally MDA 
values for rats treated with AA, AT, and TS for 8 
weeks alone were similar to control. 



Testicular oxidative stress in Sprague-Dawley rats administered Momordica charantia 

108 

Testicular ascorbic acid concentration levels 
The mean AA values followed an inverse 
relationship to MDA concentration. It showed 
a significant (p < 0.05) duration dependent 
decrease in testicular AA form a control of 0.15 
± 0.02, 0.12 ± 0.02, 0.13 ± 0.02 to 0.01± 0.02, 
0.01± 0.01 and 0.00 ± 0.01 for animals fed the 
extract for 6, 8 and 16 weeks. Also those pre-
treated with AA, AT, TS for 8 weeks and later 
post-treated with the extract for another 8 weeks 
(0.02±0.02, 0.04±0.06, 0.02±0.02) compared to 
control. The testicular AA showed substantial 

recovery to base line control value in rats fed the 
extract for 8 weeks thereafter treated with AA, 
AT and both TS for 8 weeks (0.10 ± 0.02, 0.08 
± 0.04, 0.12 ± 0.02; p < 0.05; Table-I).  
 
The group in which the extract was 
administered concurrently with AA, AT and 
TS for 8 weeks, showed an appreciable 
modulation (Table-I). These showed no 
significant difference. Lastly, AA values for 
rats treated with AA, AT, and TS for 8 weeks 
alone were similar to control. 

 
Table II: Serum Testosterone levels in experimental and control Sprague-Dawley rats  

Groups (n =75)  Testosterone (ng/ml) 

B1 
B2 
B3 

AA8wks – MC 8wks 
AT8wks–MC8wks 
AA8wks,AT8wks –MC8wks 

0.13 ± 0.02b 
0.12 ± 0.07b 
0.17 ± 0.09b 

C1 
C2 
C3 

MC8wks–AA8wks 
MC 8wks –AT8wks 
MC 8wks –AA8wks,AT8wks 

0.32 ± 0.35 
0.35 ± 0.24 
0.34 ± 0.21 

D1 
D2 
D3 

MC8wks+AA8wks 
MC8wks +AT8wks 
MC8wks+AA8wks,AT8wks 

0.31 ± 0.08 
0.34 ± 0.20 
0.37 ± 0.24 

E1 
E2 
E3 

AA8wks 
AT8wks  
AA8wks,AT8wks 

0.38 ± 0.06 
0.33 ± 0.15 
0.37 ± 010 

F1 
F2 
F3 

Distilled water6wks 
Distilled water8wks 
Distilled water16wks          

0.36 ± 0.02 
0.31 ± 0.09 
0.29 ± 0.01 

Values expressed as Mean ± standard deviation; bp < 0.05; Distilled water given for 6 to 8 weeks; 
MC8wks: 50 mg/100 g of Momordica charantia seed extract fed for 8wks; AA8wks: 0.01 of ascorbic 
acid treated for 8wk; AT8wks: 20 mg/kg of α-tocopherol treated for 8wk; AA8wks, AT8wks: α-
tocopherol & ascorbic acid at doses 0.01 and 20 mg/kg administered simultaneously for 8wk; wk: 
weeks. 
 

Testosterone concentration 
The serum TT level following administration 
of the extract for 6, 8 and 16 weeks were 
observed to diminish markedly. It decreased 
from 0.36 ± 0.02 (control) to 0.15 ± 0.09 (after 
8 weeks) and 0.05 ± 0.02 ng/ml (after 16 
weeks; Figure 1). These values only become 
significant (p < 0.05) after the 8 weeks. A 
similar reduction in serum TT levels was also 
observed in rats pre-treated prophylactically 
with AA, AT and both TS for 8 weeks 
followed by the extract for another 8 weeks 
(Table-II). There was no significant (p < 0.05) 
difference observed in serum TT in rats 

administered the extract and antioxidant 
concurrent and with TS treatment alone 
compared to control (Table-II). 
 
Discussion 

It has been shown previously from research 
that administration of AA to normal animals 
stimulates both sperm production and TT 
secretion [18]. It is also known that AA 
counteracts the testicular oxidative stress 
induced by exposure to pro-oxidant substances 
such as arsenic, cadmium, endosulfan and 
alcohol [19, 20]. In this present study, animals fed 



Yama OE, Duru FIO, Oremosu AA, Noronha CC 

109 

the extract alone (Group A) and those pre-treated 
with TS before administering the extract (Group 
B) had both showed a high level of testicular 
lipid peroxidation. This means the extract 
triggered oxidative stress via the release of free 
radicals in the testes, as evidenced by the 
elevation of testicular MDA and also a decrease 
in testicular AA in these groups. The treatment 
with AT has also been shown to suppress lipid 
peroxidation in testes [21] and reverse 
detrimental effects of oxidative stress on 
testicular function [22, 23]. This finding is in 
concert with our results, where the extract 
resulted in a decreased in testicular AA level. 
There is a high possibility that the extract may 
have acted via production of oxidative stress, in 
view of the fact that co-administration of AA, 
AT or both TS with MC (Group D) were 
found to protect against the elevation of 
MDA levels as values were identical to 
control. It was also observed that both TS and 
AA offered a better protection against the 
oxidative stress from MC than AT when 
concurrently administered with the extract 
compared to their control counterpart. The 
reason for these differential actions of these two 
vitamins cannot be fully explained. However, it 
is possible for AA to be acting in somewhat 
unexplained mechanism(s) in addition to its 
action as an antioxidant. It is known that AA is 
necessary for steroidogenesis [24] and has a co-

enzymatic function in the biosynthesis of 
steroid hormones, such as testosterone [21]. Our 
finding of a decreased serum TT level in 
Group A rats fed the extract may not be 
unconnected with the effect of MC. Studies 
have shown that the serum TT level correlates 
positively with sperm concentration and 
motility [25, 26]. It is also known that sperm 
production cannot proceed to an optimal 
completion without a continuous TT supply 27]. 
Our result demonstrated that MC produced a 
significant reduction in serum TT level and 
therefore, could be linked to a cessation of 
spermatogenesis. A resulting decreased 
spermatogenesis and increased sperm damage 
secondary to the oxidative stress induced by 
MC at the level of testicular micro-
environment could therefore be correctly 
extrapolated. Also the MDA resulting from the 
membrane damage can also induced further 
sperm damage have shown in previous studies 
[28]. Thus animals in these groups were 
expected to show a diminished fertility.  
 
In Conclusion, the present study showed that 
MC exerted its effect via generation of free 
radicals with accompanying decrease in serum 
TT, when administered at an oral dose of 
50mg/100 g body weight of rat. This effect was 
also observed to be dose dependent.  

Serum Testosterone level in Sprague-Dawley rats treated with 
Momordica charantia  extract compared to control

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

ControlWk 6 Wk 8 Wk 16

S
er

u
m

 T
es

to
st

er
o
n

e 
(n

g
/m

l)

 

 

Figure 1: Serum testosterone level in Sprague-Dawley rats treated with Momordica charantia 
extract compared to control 



Testicular oxidative stress in Sprague-Dawley rats administered Momordica charantia 

110 

Acknowledgement 

We wish to acknowledge Mr. Adeleke of the 
Department of Pharmacognosy, Faculty of 

Pharmacy, University of Lagos, Nigeria for his 
help in preparation of the herbal decoction and 
encouragement to do this work.  

 
______________ 

 
References 

1. Johnson FC. The antioxidant vitamins CRC. Crit Rev 
Food Sci Nutr 1979; 11: 217-309. 
doi:10.1080/10408397909527264.  

2. Paolicchi A, Pezzini A, Saviozzi M. Localization of a 
GSH-dependent dehydroascorbate reductase in rat 
tissues and subcellular fractions. Arch Biochem 
Biophys 1996; 333: 489-95. 
doi:10.1006/abbi.1996.0419. PMid:8809091.  

3. Ozturk A, Ballad AK, Mogulkoc IT, Ozturk B. The 
effect of prophylactic melatonin administration on 
reperfusion damage in experimental testis ischemia- 
reperfusion. Neuro Endocrinol Lett 2003; 24: 3-4.  

4. Alvarez JG, Storey BT. Lipid peroxidation and the 
reactions of superoxide and hydrogen peroxide in 
mouse spermatozoa. Biol Reprod 1984; 30: 833-41. 
doi:10.1095/biolreprod30.4.833. PMid:6329333.  

5. Sies H. Oxidative stress, Oxidants and antioxidant. 
Vol. II. London: Academic Press; 1985. p. ?.  

6. Agarwal A, Gupta S, Sharma RK. Role of oxidative 
stress in female reproduction. Reprod Biol 
Endocrinol 2005; 3: 28. doi:10.1186/1477-7827-3-28. 
PMid:16018814. PMCid:1215514.  

7. Cummins JM, Jequier AM, Kan R. Molecular biology 
of human male infertility: links with aging, 
mitochondrial genetics and oxidative stress? Mol 
Reprod Dev 1994; 37: 345-62. 
doi:10.1002/mrd.1080370314. PMid:8185940.  

8. Rao M, Narayana K, Benjamin S, Bairy KL. L-
ascorbic acid ameliorates postnatal endosulfan 
induced testicular damage in rats. Indian J Physiol 
Pharmacol 2005; 49: 331-6. PMid:16440852.  

9. Imlay JA. Redox pioneer: Professor Irwin Fridovich. 
Antioxid Redox Signal 2010. [Epub ahead of print].  

10. Technical data report for bitter melon (Momordica 
charantia). 2002; p. 5, 53-83.  

11. Kontush A, Finckh B, Karten B, Kohlschutter A, 
Beisiegel U. Antioxidant and pro- oxidant activity of 
alpha-tocopherol in human plasma and low density 
lipoprotein. J Lipid Res 37: 1436-48. PMid:8827516.  

12. Helzlsouer KJ, Huang HY, Alberg AJ, Hoffman S, 
Burke A, Norkus EP, et al. Association between α-
tocopherol, γ-tocopherol, selenium, and subsequent 
prostate cancer. J Natl Cancer Inst 2000; 92: 2018-23. 
doi:10.1093/jnci/92.24.2018. PMid:11121464.  

13. Mishra M, Acharya UR. Protective action of vitamins 
on the spermatogenesis in lead- treated Swiss mice. J 
Trace Elem Med Boil 2004; 18: 173–178. 
doi:10.1016/j.jtemb.2004.03.007. PMid:15646264.  

14. Malaka SLO. [Personal Communication]. Lagos 
2005; Department of Zoology, University of Lagos, 
Nigeria.  

15. Buege JA, Aust SD. Microsomal lipid peroxidation. 
Methods Enzymol 1978; 52: 302-10. 
doi:10.1016/S0076-6879(78)52032-6.  

16. Association of Official Analytical Chemists. 
Determination of Vitamin C. In: Holowits W. ed. 
Official methods of analysis 1990; 16: 140.  

17. Tietz NW. Clinical guide to laboratory tests. 3rd ed. 
Philadelphia: WB Saunders; 1995. p. 578-80.  

18. Sönmez M, Türk G, Yüce A. The effect of ascorbic 
acid supplementation on sperm quality, lipid 
peroxidation and testosterone levels of male Wistar 
rats. Theriogenology 2005; 63(7): 2063-72. 
doi:10.1016/j.theriogenology.2004.10.003. 
PMid:15823361.  

19. Senthil kumar J, Banudevi S, Sharmila M. Effects of 
Vitamin C and E on PCB (Aroclor 1254) induced 
oxidative stress, androgen binding protein and lactate 
in rat Sertoli cells. Reprod Toxicol 2004; 19: 201-8. 
doi:10.1016/j.reprotox.2004.08.001. PMid:15501385.  

20. Maneesh M, Jayalakshmi H, Dutta S. Experimental 
therapeutic intervention with ascorbic acid in ethanol 
induced testicular injuries in rats. Indian J Exp Biol 
2005; 43: 172-6. PMid:15782819.  

21. Lucesoli F, Fraga CG. Oxidative stress in testes of 
rats subjected to chronic iron intoxication and alpha-
tocopherol supplementation. Toxicology 1999; 132: 
179-86. doi:10.1016/S0300-483X(98)00152-8.  



Yama OE, Duru FIO, Oremosu AA, Noronha CC 

111 

22. Sen Gupta R, Sen Gupta E. Vitamin C and vitamin E 
protect the rat testes from cadmium-induced reactive 
oxygen species. Mol Cells 2004; 17: 132-9. 
PMid:15055539.  

23. Verma RJ, Nair A. Ameliorative effect of vitamin E 
on aflatoxin-induced lipid peroxidation in the testis of 
mice. Asian J Androl 2001; 3: 217-21. 
PMid:11561193.  

24. Das KK, Dasgupta S. Effect of Nickel sulfate on 
testicular steroidogenesis in rats during protein 
restriction. Environ Health Perspect 2002; 110: 923-
6. doi:10.1289/ehp.02110923. PMid:12204828. 
PMCid:1240993.  

25. Caroppo E, Niederberger C, Lacovazzi PA, 
Palaguano A, D’Amato G. Human chorionic 
gonadotropin free beta-subunit in the human seminal 
plasma: a new marker for spermatogenesis? Eur J 

Obstet Gynecol Reprod Biol 2003; 106(2): 165-9. 
doi:10.1016/S0301-2115(02)00231-2.  

26. Osinubi AA, Adeyemi A, Banmeke A, Ajayi G. The 
relationship between testosterone concentration 
sperm count and motility in fertile Nigerian males. 
Afr J Endocrinol Metab 2003; 1: 43-5.  

27. Mohri H, Suter DA, Brown-Woodman PD, White IG, 
Ridley DD. Identification of the biochemical lesion 
produced by alpha-chlorohydrin in spermatozoa. 
Nature 1975; 255 (5503): 75-7. 
doi:10.1038/255075a0. PMid:1128672.  

28. Kodama H, Yamaguchi R, Fukuda J, Kasai H, 
Tanaka T. Increased oxidative deoxyribonucleic acid 
damage in the spermatozoa of infertile male patients. 
Fertil Steril 1997; 68: 519-24. doi:10.1016/S0015-
0282(97)00236-7.  

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