ArticlePDF Available

Protective effect of combined pumpkin seed and ginger extracts on sperm characteristics, biochemical parameters and epididymal histology in adult male rats treated with cyclophosphamide



Reproductive toxicity is one of the side effects of cyclophosphamide (CP) in cancer treatment. Pumpkin seeds and Zingiber officinale are natural sources of antioxidants. We investigated the possible protective effect of combined pumpkin seed and Zingiber officinale extracts on sperm characteristics, epididymal histology and biochemical parameters of CP-treated rats. Male adult Wistar rats were divided randomly into six groups. Group 1, as a control, received an isotonic saline solution injection intraperitoneally (IP). Group 2 were injected IP with a single dose of CP (100 mg/kg) once. Groups 3 and 4 received CP plus 300 and 600 mg/kg combined pumpkin seed and Zingiber officinale extract (50:50). Groups 5 and 6 received only 300 and 600 mg/kg combined pumpkin seed and Zingiber officinale extract. Six weeks after treatment, sperm characteristics, histopathological changes and biochemical parameters were assessed. In CP-treated rats, motile spermatozoa were decreased, and abnormal or dead spermatozoa increased significantly (P < 0.001) but administration of the mixed extract improved sperm parameters. Epididymal epithelium and fibromascular thickness were also improved in extract-treated rats compared to control or CP groups. Biochemical analysis showed that the administration of combined extracts could increase the total antioxidant capacity (TAC) level significantly in groups 3, 4, 5 and 6. Interestingly, the mixed extract could decrease most of the side effects of CP such as vacuolization and separation of epididymal tissue. Our findings indicated that the combined extracts might be used as a protective agent against CP-induced reproductive toxicity.
February 2014, Volume 11, Number 1
Combined Effect of Ginger and Pumpkin Seed Extracts
on Rat Testis and Serum Biochemical Parameters after
Cyclophosphamide Treatment
Forouzan Mohammadi 1, Hossein Nikzad 1, 2 *, Aliakbar Taherian 2, Mohsen Taghizadeh 3, Abolfazl Azami-Tameh 1, Homayon Naderian 1,
Mohammad Ali Atlasi 1
1. Anatomical Sciences Research Center, Kashan University of Medical Sciences, Kashan, Iran.
2. Gametogenesis Research Center, Kashan University of Medical Sciences, Kashan, Iran.
3. Research Center for Biochemistry and Nutrition in Metabolic Disorders. Kashan University of Medical Sciences, Kashan, Iran.
* Corresponding Author:
Hossein Nikzad, PhD
Gametogenesis Research Center, Kashan University of Medical Sciences, Kashan, Iran.
Tel/Fax: +98 (31) 55621158
Introduction: Cyclophosphamide is a chemotherapy drug with several side effects on various
organs such as male reproductive system that can cause infertility. In this study, we assessed
combined effect of ginger and pumpkin extractson rat testis after CP injection.
Methods: Forty adult male rats were randomly divided into 4 groups: The control group
received intraperitoneal injection of isotonic saline solution. The cyclophosphamide (CP) group
received a single dose of cyclophosphamide (100 mg kg_1BW) intraperitoneally. Combined
extracts (ginger+ pumpkin) group received orally 300 mg combined extracts and combined
extracts(ginger+ pumpkin) +CP groups received orally 300 mg combined extracts for a period
of 6 weeks after CP injection.
Results: Our results showed that although ginger extract could not change testis weight,
testosterone, malondialdehyde (MDA) and ROS, antioxidant level in serum was increased
signicantly. Epithelium thickness and tube diameter were decreased in combined groups with
or without CP in comparison to control group. The combined extract could improve histological
changes in both combined extract and combined extract+ CP compared to CP group, which
could be attributed to the higher serum level of antioxidants.
Conclusion: The administration of combined extracts can increase the serum antioxidant level
and decrease the side effects of CP on testis.
Article info:
Received: 14 Aug 2013
Accepted: 24 Dec 2013
Key Words:
Biochemical parameters.
Hossein Nikzad is an academic member (Professor) in Kashan University of Medical Sciences and teaches anatomy and
embryology courses for medical and paramedical students for 20 years. His interest research eld is clinical embryology,
IVF, reproducon and androloy. Unl now, he published more than 40 manuscript at the naonal and internaonal jour-
nals. Now, he is head of gametogenesis research center in Kashan University of Medical Sciences.
February 2014, Volume 11, Number 1
1. Introduction
n inability to conceive after 12 months of
sexual practice without using any contra-
ception is dened as infertility[1].World
Health Organization reported that 10–
15% of young couples are faced with in-
fertility and each gender shows 50% of the related caus-
ing factors[2]. It has been reported that environmental
factors such as pesticides, exogenous oestrogens, heavy
metals and chemotherapy are the reasons for declin-
ing male sperm count which may have a negative im-
pact on male fertility[3]. cyclophosphamide (N, N-bis
(2-chloroethyl) tetrahydro-2H-1, 3, 2-oxazaphospho-
rin-2-amine2-oxide), a cytotoxic alkylating agent is
a nitrogenous mustard belonging to the group of cy-
totoxic or cytostatic drugs[4]. Studies have shown
that generation of free radicals and reactive oxygen
species isassociated with CP treatment as well. It is
known that CP disrupts the redox balance of tissues
resulting in oxidative stress. It has been reported that
oxidative DNA damage is caused by hydroperoxide
derivatives of CP through generation of H2O2.Also,
acrolein, another component of CP, has been found to
interfere with the tissue antioxidant defense system
and produces highly reactive oxygen free radicals,
which are mutagenic to mammalian cells[5].Lipid per-
oxidation has been suggested to be closely related to
CP-induced testicular damage, and malondialdehyde
(MDA) is a good indicator of lipidperoxidation that
could induce sperm abnormality[4].CP treatment is
associated with oligozoospermia and azoospermia, as
well as biochemical and histological alterations in the
testis and epididymis of human and rats [6, 7].Further-
more, disturbance in gonadotropin secretion, testicular
damage, and decreased plasma testosterone levels are
found in patients enduring treatment with CP[8]. Me-
dicinal herbs have been popular among people from
ancient times, and in recent years, a new interest has
emerged to use medicines with natural and especially
herbal origin like pumpkin and ginger [9]. Medicinal
plants contain phytochemicals and numerous chemi-
cal compounds, which can be implemented in phar-
macology by isolating the active compounds to gen-
erate new medicines and provide alternative healing
methods[10]. In most cases, herbal medicine offers
less invasive and less costly physical and emotional
treatment compared with other procedures.
Ginger rhizome (ZingiberofcinaleR., family:
Zingiberaceae) is used worldwide as a spice. Both
antioxidative and androgenic activity of ginger have
been reported in animal models[11].It contains sev-
eral compounds including acids, resins, vitamin C
compounds, folic acid, inositol, choline, pantothenic
acid,gingerol, sesquiterpene, vitamin B3 and B6 vol-
atile oils and bio-trace elements like Ca, Mg, P and
K[12]. This plant has been considered a safe herbal
medicine with few side effects[9].Ginger has been pre-
viously shown to stimulate spermatogenesis[12]. Mor-
akinyo et al. (2008)suggested that ginger extract (500
and 1000 mg kg_1B.W doses)has a benecial effect
on male reproductive functions in rats, which is con-
rmed by other studies showing an increase in sperm
count, motility,testosterone, antioxidant enzymes, su-
peroxide dismutase (SOD), catalase, glutathione per-
oxidase (GSH) and a decrease in malonhydialdehyde
lipid peroxidation levels [13, 14].We recently showed
that ginger extract at doses of 300 and 600 mg/ kg BW
has a positive effect on recovery of spermatogenesis
in adult rats after cyclophosphamide (CP) treatment.
Also, we found that co-administration of this extract
with CP can counterbalance the negative effect of CP
on testis parameters demonstrated in our study[1].
Pumpkin (cucurbitapepo var. styriaca) family cucur-
bitacea is an important leaf and seedvegetable tropical
vine grown and highly reputed in traditional medicine
and largelyconsumed in many countries such as Iran
[15]. The seeds are a rich natural source of proteins,
phytosterols, polyunsaturated fatty acids, phytochemi-
cals, sterols, antioxidant vitamins such as carotenoids
and tocopherol and trace elements such aszinc and
selenium[4].It has been demonstrated that pumpkin
seeds and daily rich diet of zinc can decrease the un-
desirable side effect of lead contaminants and improve
the sexual health status[16].Pumpkin seeds improve
sexual stimulation and intromission and ejaculatory
latency[17]. Pumpkin causes a signicant reduction in
sperm count with primary and secondary abnormali-
ties by producing further zinc and protein. Therefore,
pumpkin is proposed for both the prevention and treat-
ment of infertility in male animals[18].The ndings of
our recent study indicated that pumpkin seed extract
could recover the side effects of CP, epididymis histol-
ogy and sperm parameters through preventing oxida-
tive stress[4].
Although it is now broadly accepted that ginger and
pumpkin seeds have a positive effect on fertility, this
study is the rst study that evaluates the effects of the
combined extracts of ginger and pumpkin seeds on
CP-treated rat testis. Therefore, the present study was
designed to investigate any possible protective effects
February 2014, Volume 11, Number 1
of the combined extracts of ginger and pumpkin on
biochemical parameters and testicular histology of
CP-treated male rats.
2. Materials and Methods
Cyclophosphamide was purchased from Baxter On-
cology GmbH, Frankfurt, Germany. Pumpkin seeds
(Cucurbita Pepo var. Styriaca) were purchased from
local Iranian markets. Ginger was obtained from Nat-
ural Remedies Company in India that has been stan-
dardized as ‘total Gingerol 5%’.MDA, ROS, antioxi-
dant and testosterone kits were purchased from Glory
Science Co. Ltd, China.
Animal experiment :In this study, forty healthy adult
male Wistar rats (8–10 weeks old, 300–350 g) were
obtained from Kashan University of Medical Sciences.
Rats were in wire-wooden cages under controlled light
schedule (12 h light and 12 h darkness). The animals
were allowed to acclimatize for a period of 7 days be-
fore starting the experiment. During the treatment pe-
riod (Six weeks) they were fed with the supplied food
pellets and had free access to water. All experiments
were implemented in accordance with the guidelines
and were approved by the Local Committee on Animal
Research in Kashan University of Medical Sciences.
Study design and treatment: The rats were randomly
divided into four groups of ten. Control group received
a single intraperitoneally injection of isotonic saline
solution (1 ml). CP Group received a single dose of
cyclophosphamide (100 mg kg_1body weight)intra-
peritoneally[19].Group ginger +pumpkin 300received
300 mg/kg BW of ginger extract plus pumpkin seed
extract. Group CP+ginger +pumpkin 300 received CP
plus300 mg/kg BW ginger and pumpkin seed extract
orally for a period of 6 weeks after CP injection.
Food regimens: The synthetic diet was purchased
from Behparvar Company in Iran. Pumpkin seed with-
out oil (cold pressed oil) was obtained from Barij Es-
sence Company in Kashan, Iran. The pumpkin seed
without oil was mixed with 70% alcohol in a blender
and incubated for 72 hours at room temperature. Then
ethanol liquid was separated with a lter paper. The
extract was standardized to contain amino acid, total
avonoid and total phenolic compound concentra-
tion[20, 21]. One kg of the synthetic diet was mixed
with 512cc of pumpkin seed extract (2.6%) for 300
mg/kg body weight dosage. Also, ginger extract was
dissolved in 50% ethanol and with pumpkin seed ex-
tract, was added to powdered food pellets of rats. The
mixture was dried in oven under 50°C and stored at
4°C. The animals were fed based on a daily consump-
tion of 15-17 gr of dried diet/rat. Before starting the
experiment, we measured the food intake in control
and CP-treated rats. In case of CP-injected rats, the
food intake was decreased to about 8 grams in the rst
5 days after injection. So, the ginger+ pumpkin content
of the food was adjusted to give the right dose during
the experiment. Fresh diet was prepared weekly.
Sample collection: Since spermatogenic period in
rats is 48 days[22], in our study, rats were weighed and
killed under anaesthesi a 42 days after CP injection.
Blood samples were collected from the left ventricle,
and plasma was immediately separated for biochemi-
cal examinations. After weighing, testis was xed with
Bouin’s solution for histology.
Biochemical studies: The level of rat testosterone, to-
tal antioxidant capacity (T-AOC), MDA and reactive
Parameters Control CP Combined Extract 300 CP+300 Combined Extract
Tube diameter(µm) 286.20±13.74 270.80±19.81 253.18±35.6* 245.57±18.72*
Epithelium thickness (µm) 152.79±7.21 131.16±6.60† 114.60±6.50† 111.42±2.8†
Lumen diameter(µm) 133.41±7.93 139.65±18.21 139.99±31.02 134.14±18.20
*P < 0.05 compared with control group.
†P < 0.001 compared with control group.
Table 1. Administration effect of the combined extracts of ginger and pumpkin seeds on histological parameters in rat
seminiferous tubes.
February 2014, Volume 11, Number 1
oxygen species were determined using an enzyme-
linked immunosorbent assay (ELISA) method. Blood
was collected from left ventricle of anaesthetized rats
in a tube and centrifuged Immediately at 3000 rpm for
10 min. The separated serum was deposited in -20 °C
freezers for the biochemical tests. The biochemical
tests were carried out according to the kit instruction.
Briey, a series of standard solutions were prepared.
First, the primary antibody was added to the sample
wells of a 96-well plate (not to control or standard
wells). 40 ul of samples or 50 ul of standard solutions
were added to the designated wells. After adding the
secondary antibody (labelled with Strep-tavidin-HRP)
to the standard and sample wells, the plate was incubat-
ed at 37 °C for 60 min. Plate was washed;chromogenic
solutions (A, B) were added and incubated at 37 °C in
the dark for 10 min. To the control wells, only chro-
mogenic and stop solutions were added. After adding
the stop solutions to the wells, the absorbance of wells
at450 nm wavelengths was read by an Elisa Reader
(ModelStatFax 2100). A standard curve was extrapo-
lated based on the readings of the standard wells, and
linear regression equation was calculated. The concen-
tration levels of samples were calculated based on OD
readings of the samples and the standard curve.
Histology and light microscopy: The left testis was
carefully dissected, trimmed of all fats and blotted dry
to remove any blood. The testis was divided into three
parts, and the middle section was xed in Bouin’s uid
for 48 h. The xed samples were dehydrated in graded
levels of ethanol, cleared in xylene and embedded in
parafn wax for sectioning. Five μm thick sections
were prepared and stained with Hematoxylin and Eo-
sin (H&E) and observed under a light microscope.
Morphometric study:An optical microscope (Zeiss/
German) with an objective lens ×40 was used for cell
counting. Spermatogonia, primary spermatocyte, sper-
matid, spermatozoa and Sertoli cells were counted in
10 seminiferous tubules in stages VII or VIII in each
animal. The diameter of the round or nearly round
seminiferous tubules was estimated as the average of
two perpendicular longer and shorter diameters. Epi-
thelium thickness was obtained by the same method.
The size of the lumen was calculated by subtracting
epithelium thickness from the diameter. A mean value
of each of these parameters was calculated for each
group and was compared with the other groups.
Statistical analyses: The data are presented as mean
± SEM (standard error of mean). Statistical analyses
were carried out using ANOVA test and a P value of P
<0.05 was considered as statistically signicant.
3. Results
Morphometry: Testicular weight did not change sig-
nicantly between the control and other groups. The
general effects of CP were some hair loss and a de-
creased appetite in the rst5 days after CP injection
only seen in CP group. CP treatment decreased the
number of germ cells as well as epithelium thickness of
seminiferous tubules (Tables 1 and 2). Other features
Parameters Control CP Combined Extract 300 CP+300 Combined Extract
Spermatogonia(n) 63.15±8.15 51.55±3.51† 64.17±4.8†† 61.50±5.04††
Spermatocytes(n) 74.20±11.16 57.24±2.15* 71.03±6.5†† 71.41±4.5††
Spermad(n) 230.0±38.27 196±4.5 211.9±15.7 201.74±13.9
Sperm(n) 180.07±5.5 161.11±4.6†† 190.2±9.5*†† 175±4.57††
Sertoli(n) 17.75±3.89 15.22±2.99 18.82±2.81 18.56±1.47
*P < 0.05 compared with control group.
†P < 0.001 compared with control group.
††P < 0.001 compared with CP group.
Table 2. Administration effect of the combined extracts of ginger and pumpkin seeds on cell count of germ and Sertoli cell
parameters in rat seminiferous tubes.
February 2014, Volume 11, Number 1
of the CP-treated seminiferous tubules were exfolia-
tion of germ cells and vacuolated appearance of the
epithelium. The administration of combined extract to
CP-treated or normal rats could signicantly increase
germ cells count in seminiferous tubules (spermato-
gonia, spermatocytes, sperm) compared to CP group.
However, epithelium thickness and tube diameter were
decreased in combined groups with or without CP in
comparison to control group.
Biochemical Parameters
Antioxidant levels: Cyclophosphamide treatment
did not change the antioxidant level signicantly in
comparison with the control group. However, admin-
istration of the combined extracts 300 mg kg_1 BW_1
strongly increased antioxidant levels compared with
that of control or CP group (Table 3).
Testosterone, ROS and MDA: Testosterone, Ros and
MDA levels did not change signicantly in the differ-
ent groups (Table 3).
Histology: Cyclophosphamide treatment caused
a reduction in the size, epithelium thickness and the
number of different types of cells in the seminiferous
tubules. Degeneration, vacuolation and exfoliation of
germ cells into the lumen of seminiferous epithelium
were other features of the CP group samples. However,
administration of the combined extracts300 mg kg_1
BW_1to CP group rats and rats without CP caused an
improvement in the germ cells count of the seminifer-
ous tubules compared with the CP group but epithe-
lium thickness and tube diameter decreased in com-
parison to control group in these groups.
4. Discussion
Using chemotherapy drugs like CP for cancer treatment
is limited by their side effects. The side effects of che-
motherapy include reproductive toxicity that has been
documented in different studies[19].This study was
completed to investigate the effects ofthe combined
extracts of ginger plus pumpkin seeds on CP-injected
and normal rat testis. To our knowledge, this is the
rst study investigating the effects of this combined
extract against testicular damage caused by CP in rats.
Our results showed that in combined extract-treated
rats (300 mg kg_1BW_1), the number of germ cells
in seminiferous tubules was increased signicantly in
comparison to CP group alone. This result reconrms
our last report indicating that administration of ginger
or pumpkin increases the number of germ cells in sem-
iniferous tubes and has a positive effect on recovery of
spermatogenesis in adult rats after cyclophosphamide
(CP) treatment[1, 23].
Epithelium thickness and tube diameter decreased
in comparison to control group in these groups. Our
results support those of[24] Saalu reported decrease
in tubular diameter and epithelium thickness after ad-
ministration of uted pumpkin extract 400 mg/kg/day/
oral and our result in another study where we found
that administration of ginger extract 300 mg/kg/day/
oral decreased epithelium thickness in comparison to
control group [1].
In our study, the weight of the testis did not change
signicantly between the different groups. A decreased
testis weight has been reported in CP-treated rats [25].
In that study a dose of 15 mg kg_1 BW of CP was
Parameters Control CP Combined Extract 300 CP+300 Combined Extract
MDA(nmol/ml) 2.42±0.46 2.71±0.42 2.05±0.56 2.50±0.40
ROS 3.40±0.85 2.60±1.03 4.17±0.67 4.24±0.56
Anoxidant(µl) 5.51±3.52 5.20±2.43 11.97±3.6*†† 11.92±3.22*††
Testosterone(ng/dl) 7.38±2.09 8.39±1.43 9.45±1. 93 2.06 ±7.93
*P < 0.05 compared with control group.
††P < 0.001 compared with CP group.
Table 3. Administration effect of the combined extracts of ginger and pumpkin seeds on serum levels of testosterone, anti-
oxidant, Malondialdehyde (MDA) and ROS in rat seminiferous tubes.
February 2014, Volume 11, Number 1
given to rats by oral gavage once a week for 10 weeks
(in total 150 mg kg _1BW_1), while in our study, 100
mg kg_1 BW_1 of CP was injected in a single dose.
The chronic low-dose administration of CP to male
rats could be the reason for decreased reproductive or-
gan weights[26]. In another study, two different doses
of CP (100 and 200 mg kg_1) were injected to male
rats. A decrease in the weight of testis was detectable
1 week after the injection for both doses, but after 5
weeks, the reduced weight of testis was only detect-
able in 200 mg kg_1injected rats[27]. In our study, the
samples were collected and studied 6 weeks after the
injection of 100 mg kg_1 of CP; therefore, we could
not see any difference in testis weight. Of course, the
reason that samples were studied after 6 weeks was
that spermatogenesis (development of mature sperma-
tozoa from diploid spermatogonial cells) in rats takes
48 days[22].
In CP treated rats the toxic effect was indicated by
signicant reduced spermatogonia,spermatocyte and
sperm count. Our results support those studies report-
ing irregular and diminished seminiferous tubules
containing only a few germ cells in the CP group[19].
Our results provided no evidence for those studies
[19]which reported that MDA and ROS levels were
increased signicantly in CP-treated rats. Since we ex-
amined the rats 42 days after chemotherapy, probably
the levels of these parameters have been recovered in
the rst or second weeks of the experiment. This re-
covery most probably has been done because of the
effective help of the mixed extract.In our study, tes-
tosterone, ROS and MDA levels neither changed sig-
nicantly in CP group nor in combined extract -treated
groups (with and without CP).
Our results showed that the combined extract with
dose (300 mg kg_1 BW_1) could increase antioxidant
to a higher level. This result conrms the antioxidant ef-
fects of Telfairia occidentalis (uted pumpkin) extracts
reported by Nwanna and Oboh[28] and zingiberreport-
ed by Morakinyo[12]. Antioxidant therapy improves
fertility parameters through a protective mechanism
against oxidative stress[11].Phenolic compounds such
as polyphenols avonoids as well as vitamins and zinc
in medicinal plants such as ginger and pumpkin are
attributed factors for antioxidant activity [4, 12]. In
studies by Tsai et al. (2006) and Gossell-Williams et
al.(2006), it was reported that Pumpkin seed oil is rich
in many powerful antioxidants and useful nutritional
supplements such as essential fatty acids and polyun-
saturated fatty acids including linoleic acid,oleic acid,
palmitic acid, omega 3, 6 and 9, carotenes, lutein,
gamma and P-tocopherols,phytosterols, chlorophyll,
selenium and zinc[29, 30].Also, the presence of oleic
acid, amonounsaturated fatty acid in pumpkin reduces
the susceptibility of the testis and epididymis to lipid
Peroxidation[31].Zinc in pumpkin seeds is an essen-
tial trace mineral that acts as an antioxidant by neu-
tralizing free radical generation. Also,Zinc could play
a direct antioxidant action by engrossing the iron or
copper binding sites of lipids, proteins, and DNA[14].
On the other hand,all major active ingredients of Z.
ofcinale such as Zingerone, Ginger¬diol, Zingibrene,
gingerols and shogaols, have antioxi¬dant proper-
ties[9]. Besides, other studies have shownthat ginger
oil has a protective effect on DNA damage against Hy-
drogen Peroxide (H2O2) and might decrease oxygen
radical and could be used as an antioxidant[32, 33].
In previous studies, we showed that the higher dose
of ginger extract (600 no 300mg kg_1 BW_1) alone
could increase testosterone level. Because only the
higher dose of ginger could induce the higher level of
testosterone. Ginger extract might act in a dose-depen-
dent manner[1].In our previous work, only the lower
dose (300 mg/kg) of pumpkin seed showed antioxidant
activity but not the higher dose (600 mg/kg), which
even increased serum free radical level. Based on these
results, the combined extract was prepared from the
lower dose of each plant. Since in Iranian family gath-
erings normally both pumpkin seeds and ginger a-
vored tea are served, we decided to study the effects
ofthe combined extract on testis after chemotherapy.
The combined extract of ginger and pumpkin (300
mg kg_1 BW_1) has an antioxidant activity and thus
can reduce the adverse effects of CP in testis. Also,
these results suggest further studies to evaluate the use
of the combined extracts of ginger and pumpkin as a
supplement drug to counterbalance the negative effect
of CP in human as well.
This research is nancially supported by the Deputy
for Research of Kashan University of Medical Sci-
ences (Kaums) and Kashan Anatomical Sciences Re-
search Center (Grant No 9110).
February 2014, Volume 11, Number 1
1. Mohammadi F,Nikzad H,Taghizadeh M, Azami-Tameh A,
Hosseini M. Protective effect of Zingiber ofcinale extract
on rat testis after cyclophosphamide treatment. Androlo-
gia. 2014. 46: 680-686.
2. Kashani HH, Malekzadeh Shiravani S,Hoshmand F.The
effect of aqueous extract of Salep prepared from root-tu-
bers of Dactylorhiza maculate (Orchidaceae) on the testes
and sexual hormones of immature male mice. Journal of
Medicinal Plants Research. 2012. 6(24): 4102-4106.
3. Lamfon HA. Effect of fenugreek seed extract on carben-
dazim-inhibited spermatogenesis in albino rats. Journal of
Applied Pharmaceutical Science. 2012. 2(04): 09-13.
4. Aghaei S, Nikzad H, Taghizadeh M, Azami Tameh A,
Taherian A. Protective effect of Pumpkin seed extract
on sperm characteristics, biochemical parameters and
epididymal histology in adult male rats treated with Cy-
clophosphamide. Andrologia. 2014. 46: 927-935.
5. Rezvanfar M,Sadrkhanlou RA. Protection of cyclophos-
phamide-induced toxicity in reproductive tract histology,
sperm characteristics, and DNA damage by an herbal
source; evidence for role of free-radical toxic stress. Hum
Exp Toxicol. 2008. 27(12): 901-910.
6. Masala A, Faedda R, Alagna S, Satta A, Chiarelli G, Rova-
sio PP. Use of testosterone to prevent cyclophosphamide-
induced azoospermia. Ann Intern Med. 1997. 126: 292-295.
7. Kirkland R, Bongiovanni AM, Cornfeld D, McCormick JB,
Parks JS.Gonadotropin responses to luteinizing releasing
factor in boys treated with cyclophosphamide for nephrot-
ic syndrome. J Pediatr. 1976. 89: 941-944.
8. Hoorweg-Nijman JJ.Cyclophosphamide-induced distur-
bance of gonadotropin secretion manifesting testicular
damage. Acta endocrinologica. 1992. 126(2): 143-148.
9. Mohammadi F, Nikzad H,Taherian A,Amini Mahabadi J,
Salehi M. Effects of Herbal Medicine on Male Infertility.
Anatomical Sciences. 2013. 10(4): 3-16.
10. Erasmus N, Solomon M,Fortuin K, Henkel R.Effect of
Eurycoma longifolia Jack (Tongkat Ali) extract on human
spermatozoa in vitro. Andrologia. 2012. 44: 308-314
11. Khaki A, Fathiazad F, Nouri M,Afshin A, Hamadeh
DVM.The effects of Ginger on spermatogenesis and sperm
parameters of rat. Iranian Journal of Reproductive Medi-
cine. 2009. 7(1): 7-12.
12. Morakinyo OA, Achema PU, Adegoke OA.Effect of Zin-
giber Ofcinale (Ginger) on Sodium Arsenite Induced Re-
productive Toxicity in Male Rats. African Journal of Bio-
medical Research. 2010. 13(1): 39-45.
13. Morakinyo A, Adeniyi OS, Arikawe AP.Effects of Zin-
giber ofcinale on reproductive functions in the male rat.
African Journal of Biomedical Research. 2008. 11(3): 329-
14. Amara S, Abdelmelek H, Garrel C, Guiraud P, Douki
T.Preventive effect of zinc against cadmium-induced ox-
idative stress in the rat testis. J Reprod Dev. 2008. 54(2):
15. Nkang A,Omokaro D,Egbe A, Amanke G.Variations in
fatty acid proportions during desiccation of Telfairia oc-
cidentalis seeds harvested at physiological and agronomic
maturity. African Journal of Biotechnology. 2004. 2(2): 33-
16. Abd El-Ghany M, Dalia A, Hafez Soha M. Biological
study on the effect of pumpkin seeds and zinc on repro-
ductive potential of male rats. Faculty of Specic Educa-
tion Mansoura University,Egypt April. 2010: 14-15.
17. Gundidza G, Mmbengwa VM,Magwa ML , Ramalivhana
NJ, Mukwevho NT.Aphrodisiac properties of some Zim-
babwean medicinal plants formulations. African Journal of
Biotechnology. 2009. 8(22): 6402-6407.
18. Oyeyemi, M., S. Olukole, and O. Esan Sperm morpho-
logical studies of West African Dwarf Bucks treated with
pumpkin plant (Cucurbita pepo). Int J Morphol, 2008. 26:
19. Ilbey YO, Ozbek E, Simsek A, Otunctemur A, Cek-
men M,Somay A.Potential chemoprotective effect of mela-
tonin in cyclophosphamide-and cisplatin-induced testicu-
lar damage in rats. Fertil Steril. 2009. 92(3): 1124-1132.
20. Tsantili E, Shin Y, Nock J, Watkins C.Antioxidant con-
centrations during chilling injury development in peaches.
Postharvest Biol Technol. 2010. 57(1): 27-34.
21. Djeridane A, Yous M, Nadjemi B, Boutassouna D,
Stocker P.Antioxidant activity of some Algerian medici-
nal plants extracts containing phenolic compounds. Food
Chem. 2006. 97(4): 654-660.
22. George j.The handbook of experimental animals, The
laboratory rat. 2000, San Diego, Calif. London: Academic.
23. Mohammadi F, Nikzad H, Taghizadeh M, Moravveji
SA.Effect of pumpkin extract regimen on testicular struc-
ture and serum biochemical parameters in cyclophospha-
mide-treated adult rats. KAUMS Journal (FEYZ). 2013.
17(5): 438-446.
24. Saalu L, Kpela T, Benebo AS, Oyewopo AO, Anifowope
EO, Oguntola JA.The Dose-Dependent Testiculoprotective
and Testiculotoxic Potentials of Telfairia occidentalis Hook
f. Leaves Extract in Rat. International Journal of Applied
Research in Natural Products. 2010. 3(3): 27-38.
25. Selvakumar E, Prahalathan C,Sudharsan P, Varalakshmi
P.Chemoprotective effect of lipoic acid against cyclophos-
phamide-induced changes in the rat sperm. Toxicol. 2006.
217(1): 71-78.
26. Das UB, Mallick M, Debnath J, Ghosh D.Protective effect
of ascorbic acid on cyclophosphamide-induced testicular
gametogenic and androgenic disorders in male rats. Asian
J Androl. 2002. 4(3): 201-208.
27. Elangovan N, Chiou T, Tzeng W.Cyclophosphamide
treatment causes impairment of sperm and its fertilizing
ability in mice. Toxicol. 2006. 222(1): 60-70.
28. Nwanna EE, Oboh G.Antioxidant and Hepatoprotective
Properties of Polyphenol Extracts from Telfairia occiden-
taux (Fluted Pumpkin) Leaves on Acetaminophen Induced
Liver Damage. Pakistan Journal of Biological Sciences.
2007. 10(16): 2682-2687.
February 2014, Volume 11, Number 1
29. Gossell-Williams M, Davis A, O'Connor N.Inhibition
of testosterone-induced hyperplasia of the prostate of
sprague-dawley rats by pumpkin seed oil. Journal of Me-
dicinal Food. 2006. 9(2): 284-286.
30. Tsai YS, Tong Y, Cheng J, Lee C.Pumpkin seed oil and
phytosterol-F can block testosterone/prazosin-induced
prostate growth in rats. Urologia Internationalis. 2006.
77(3): 269-274.
31. Bourre JM, Dumont O, Durand G. Dose-effect of dietary
oleic acid: oleic acid is conditionally essential for some or-
gans. Reproduction Nutrition Development. 2004. 44(4):
32. Khaki A, Iraj S, Bazi P,Imani SAM.Comparative Study of
Aminoglycosides(Gentamicin & Streptomycin) and Fluo-
roquinolone (Ooxacin) Antibiotics on Testis Tissue in
Rats: Light and Transmission Electron Microscopic Study.
Pak. J. Med. Sci. 2009. 25(4): 624-629.
33. Grzanna R, Lindmark L, Frondoza CG. Ginger-an herbal
medicinal product with broad anti-inammatory actions.
Journal of medicinal food. 2005. 8(2): 125-132.
... A drop of the prepared sperm suspension was placed on a slide and studied under a light microscope. To estimate the percentage of motile sperms in each sample, at least 10 microscopic fields were examined at 400X magnification for each sample (20). ...
... A Neubauer slide was used to count the number of sperms in each sample after diluting the sperm suspension in formalin 10% (20). ...
... The findings of several previous studies are consistent with the current results. In this regard, Aghaei et al. (2016) demonstrated that in cyclophosphamidetreated rats, histopathological changes in the epididymis, such as vacuolization, disorganization, and separation of the epididymal epithelium, as well as sperm parameters, ameliorated remarkably by consumption of pumpkin seed extract (20). Moreover, Jalili et al. (2018) found that administration of Falcaria vulgaris extract could increase sperm parameters, such as sperm motility, count, and viability; it could also ameliorate histological changes of the epididymis in diabetic rats (23). ...
Background: In diabetic patients, uncontrolled blood sugar causes disorders in various systems of the body in the long term. The reproductive system is one of these susceptible systems. It is known that diabetes can adversely affect spermatogenesis. The use of medicinal plants in the treatment of various diseases has been discussed by many researchers for a long time. Onion, scientifically known as Allium cepa L., contains antioxidants. Insulin is also a drug used to control blood sugar in diabetic patients. Objectives: This study aimed to evaluate and compare in vivo antidiabetic activities of hydroalcoholic onion seed extract together with insulin in diabetic rats. Methods: In this study, diabetes was induced in rats with streptozotocin (60 mg/kg). Fifty animals were equally divided into five groups: nondiabetic control (group 1); diabetic control (group 2); diabetic rats receiving streptozotocin plus insulin (group 3); and diabetic rats treated with 200 and 400 mg/kg of Allium cepa L. seed extract by gavage for four weeks (groups 4 and 5). At the end of the study, the prostate ventral lobe was removed and processed for histological studies. Next, sperm parameters from the tail of the left epididymis, biochemical parameters, and histopathological changes were analyzed and compared. Results: The sperm parameters of diabetic rats receiving 200 and 400 mg/kg of Allium cepa L. extract showed a significant increase compared to the diabetic control group. Conclusions: Administration of Allium cepa L. extract as a strong antioxidant was adequate to compensate for the toxic effects of streptozotocin and increase the motility of sperms.
... Furthermore, Aghaie et al., [39] reported that Cyclophosphamide is a cancer treatment drug that is unsafe for reproduction systems. Rathinavelu et al., [40] study on the in vitro cytotoxic effects of pumpkin seed ethanolic and aqueous extracts on prostate cancer showed that these effects were mediated through oxidative stress, mitochondrial depolarization, and apoptotic pathways. ...
Full-text available
Pumpkin, belongs to the family Cucurbitaceae and genus Cucurbita, is gaining appeal across the world for a number of reasons. Pumpkin seeds have always been discarded as waste even with containing essential pharmaceutical micro and macro constituents such as proteins, antioxidative phenolic compounds, tocopherols, triterpenes, saponins, phytosterols, lignans, and carotenoids as well as these compounds D-chiro-inositol, trigonelline, and nicotinic acid. Pumpkin seeds are also rich in fibre, polyunsaturated fatty acids, vitamins, and minerals including zinc, iron, magnesium, calcium, manganese, and copper that may be used in the food industry. Pumpkin seeds are now generally used in traditional medicine in treatment of many diseases, including hypertension, rheumatoid arthritis, hyperglycemia, inflammation, dyslipidemia, bacteria, fungal infections, and tumours. As a result, pumpkin seeds are now often utilised as a herbal treatment or health-improving agent for both people and animals, and food scientists are providing the pumpkin-infused products to the food and health industries. Hence, food manufacturers have worked to broaden the range of appetiser, baking, and snack uses for pumpkin seeds. This review article provides insights into the pharmacological activities of pumpkin seeds and the possible processes which, might reduce the chance of a wide range of problems.
... Here, arrow with * indicates ''tubular atrophy with disarrangement of lining epithelial structure; arrow with ** indicates lack of sperm within the tubules.". C and 5-ASA can be compared with the findings that reported a protective effect of two antioxidants on the histology of epididymis (Aghaie et al., 2016). The results revealed that vitamin C and 5-ASA may exert their antioxidative property to prevent ACR-induced reproductive damage and their combined application is highly recommended. ...
Full-text available
Context: Serious health risks have been connected to ongoing, escalating exposure to environmental toxins and one of them is acrylamide (ACR), an organic compound. Although there are many published reports on ACR toxicity, limited information is available regarding the use of two potential antioxidants against ACR-instigated reproductive toxicity. Aims: The study focused on investigating the protective effects of vitamin C and 5-ASA against ACR-incited reproductive toxicity. Methods: A total of 50 male mice aged 4 weeks old were treated for 90 days with different concentrations either of ACR or ACR and vitamin C or ACR and 5- ASA or ACR, vitamin C, and 5- ASA. Key results: ACR significantly reduced serum testosterone level (p = 0.0037), sperm concentration (p = 0.0004), and percentage of sperm motility (p = 0.003), as well as increased sperm abnormality; head (p = 0.0058), tail (p = 0.001), and midpiece (p = 0.0339). Besides, the weight (p = 0.0006) and length (p = 0.0105) of testes, as well as weight (p = 0.0001) and length (p = 0.0021) of epididymis were decreased along with atrophy of seminiferous tubules of the testis, and disintegration of the tubular epithelium of epididymis on ACR exposed mice which were improved by vitamin C and 5-ASA administration. Conclusions: Vitamin C and 5-ASA can potentially mitigate the negative effects of ACR on male reproduction; however, combined application is recommended for better performance. Implications: In Bangladesh, this work is anticipated to address the health benefits of vitamin C and 5-ASA, particularly in improving the reproductive health of males against ACR toxicity.
... Also, proinflammatory cytokines, primarily TNF-alpha (tumor necrotic factor alpha) and interleukin 6 (IL-6) [21,22], increase in heart damage. Levels of MDA, SOD, CAT, GPx, GSH as well as total antioxidant capacity (TAC) can be measured to monitor oxidative stress [23]. MDA blood level can be measured in both humans and animals, while the activity of antioxidant enzymes is mainly analyzed from tissues, which makes it difficult to apply this method in the human population. ...
Full-text available
Introduction: Doxorubicin is an antibiotic from the anthracycline group, with clinical use limited by adverse reactions, primarily cardiotoxicity. Material and Methods: This article provides an overview of therapeutic and toxic doses of doxorubicin, the mechanism of side effects, markers for early detection as well as currently available preparations for preventing its toxicity. We searched PubMed, Google Scholar, SCIndex, Dimension, Scopus and Google for English and Serbian language abstracts, using the searching terms "doxorubicin", "cardiotoxicty", "carotenoids", "oncology", "oxidative stress", "DNA damage" and "biomarkers". Topic: The mechanism of side effects is still unclear and is considered to be multifactorial including ROS overproduction, reducing levels of endogenous antioxidants, DNA damage, large drug accumulation in cardiac tissue, calcium overload, histamine release, and impairment of autoimmune regulation of cardiac function Manifestations of cardiotoxicity are mainly acute (appear inside 24h atypical changes of ST segment, decrease in QRS complex voltage, tachycardia and supraventricular extrasystoles are observed, but can also be subacute and chronic (cardiomyocyte edema, disorganzation, fibroblast proliferation, necrosis). Diagnosis of cardiotoxicity is based on ECG, ECHO, and biochemical markers, among which the most important are troponins, while pathohistological verification is necessary for the final diagnosis. Some medications (carvedilol, atorvastatin) have showed some level of cardioprotection against DOX, but there is no overall agreement on their administration solely for this purpose. An increasing number of studies have tested various dietary supplements and natural preparations (already in the human diet) in order to discover those that could completely prevent or reduce the toxic effects of doxorubicin, with special focus on carotenoids. Conclusion: Cardiotoxicity is the leading side effect of doxorubicin, and therefore there is an active search for either new biomarkers and/or diagnostic protocols that would detect toxicity in time, as well as substances able to prevent the occurrence or alleviate DOXinduced cardiotoxicity.
... The nitric oxide rummaging action of green tea have been concentrated by [29], who inferred that specific tannins had the capacity to show magnificent cell reinforcement movement. The outcomes from the cell reinforcement properties of the pumpkin seed oil showed the compound attributes of the oil, which may represent its detailed restorative advantages [30]. ...
Full-text available
The seeds of fluted pumpkins are usually eaten boiled or pulverized and taken as food additive locally. As a result of this manner of utilization by the locals, this study was conducted to ascertain the contents of the total tocopherol, phenolic and the antioxidative property present in the oil of the seed. The pods of the fluted pumpkin were gotten from a farm within the Benin metropolis, Nigeria. The seeds were ground in a blender and afterward exposed to soxhlet extraction. Other standard methods were employed in this study. The quantitative analysis of these parameters showed Total Tocopherols (76.0 mg/kg ± 3.11), Total phenolic (3.34 mg of garlic acid equivalent ± 0.15). Percentage inhibition of DPPH was recorded to be 47.95 % ± 3.47, Nitric oxide scavenging capacity was 56.47 % ± 2.37 and ABTS was 51.06 % ± 2.60 respectively. The findings revealed that the seed oil of T. occidentalis possessed beneficial antioxidant properties which could be harnessed in management of disease like diabetes and cancer among others.
... Phenolic compounds of Zingiber were found to scavenge superoxide anion and to inhibit lipid peroxidation induced by active oxygen species such as superoxide anion or hydroxyl radical, respectively [32]. This component improved sperm parameters in adult male rats treated with cyclophosphamide [33]. It was reported that other components in honey compound syrup, such as cinnamon and saffron, showed protective effects on the reproductive system. ...
Full-text available
Objective: To evaluate the protective effects of honey compound syrup on sperm count and testis tissue in rats. Methods: Thirty rats were randomly assigned to five groups. The control group received 1 mL normal saline with dimethyl sulfoxide intraperitoneally; the busulfan group received busulfan 10 mg/kg body weight at the first and twenty-first days of the experiment via intraperitoneal injection; the last three groups received busulfan 10 mg/kg body weight to induce azoospermia, and then received 1.0, 1.5, or 2.0 mg/kg honey compound syrup, respectively, after induction of azoospermia. After administration, the testis and epididymis of all rats were removed. Then, reproductive organ weight and sperm parameters (sperm concentration, epididymal sperm reserve and daily sperm production) were measured. After hematoxylin-eosin staining, seminiferous tubule cells and diameters were assessed. Results: Busulfan damaged the testis tissue and impaired spermatogenesis. Administration of honey compound syrup in three doses improved testis tissue and spermatogenesis. The protective effects of honey compound syrup may relate to the antioxidant properties of honey and other compounds in this syrup. Conclusions: Administration of honey compound syrup could be an ameliorative agent for the side effects of chemotherapy drugs such as busulfan on the male reproductive system.
... The increased sperm count and the decreased DNA fragmentation were observed by supplementing PSO in the human trial (Elfiky et al., 2012). Aghaie et al. (2016), showed that PSO improved semen parameters in adult male Sprague-Dawley rats. Vitamin E is applied to the poultry diet to prevent from the lipid peroxidation caused by PUFAs (Esmaeili et al., 2014). ...
Full-text available
This study evaluates the effects of pumpkin seed oil and sunflower oil along with vitamin E on the reproductive characteristics of aged roosters. Thirty Ross breeder roosters (45-wk-old) were assigned into 6 groups (5 birds/group) with the following diets: 1) control group (basal diet), 2) basal diet with 2% pumpkin seed oil as omega-3 fatty acid (PSO group), 3) basal diet with 2% sunflower oil as omega-6 fatty acid (SFO group), 4) basal diet with 200 mg/kg vitamin E (Control+vitE group), 5) basal diet and 2% pumpkin seed oil along with 200 mg/kg vitamin E (PSO+vitE group) and 6) basal diet and 2% sunflower oil along with 200 mg/kg vitamin E (SFO+vitE group). Roosters were fed on the experimental diets for 60 days and different characteristics of sperm characteristics including routine semen analysis and several sperm functional tests in every 20 days were examined. Reproductive hormones were also evaluated in 0 day and at the end of the trial. Semen volume and morphology were not affected by any of the diets. The roosters fed with pumpkin seed oil+vitE showed the higher percentage of sperm concentration, total motility, progressive motility, viability and membrane integrity and the lower lipid peroxidation (P≤0.05). The group 5 (PSO+vitE) had numerically the lowest sperm with fragmented DNA (DNA Fr⁺) at day 0 of the experiment and sperm with non-fragmented DNA (DNA Fr⁻) was lowest in group 6 (SFO+vitE) on day 40 the experiment. Testosterone level was not affected by the experimental diets (P>0.05), however other hormones (LH and FSH) were affected. Based on the results, the supplementation of aged roosters’ diet with pumpkin seed oil+vitE improves reproductive performance which can be an appropriate strategy to preserve the reproductive performance of aged roosters.
... Thus, due to increasing infertility because of aging in some couples, using an antioxidant of plant origin which can have enhancing effects on sperm fertility without causing side effects seems necessary. Today, it has been found that many plants have antioxidant properties [2,9,30]. One of these plants is Ceratonia siliqua L., which is proved to reduce the adverse effects of cyclophosphamide-induced oxidative stress on sperm parameters [5,21,23]. ...
This study was conducted to determine the effects of Ceratonia siliqua L. (CS) extract on sperm parameters and DNA damage in adult male mice treated with cyclophosphamide (CP). Based on an initial dose response experiment on Ceratonia siliqua L. extract, five treatment groups were set up: control, sham (normal saline: 0.2 ml per day, IP), CP (15 mg kg-1 per week; IP), Ceratonia siliqua L. (100mg l-1 per day; IP), and group of Ceratonia siliqua L. along with CP for 35 days. After euthanizing the animals, sperms from caudal part of epididymis were collected, and their parameters, Malone Di-Aldehyde (MDA) level, and DNA fragmentation were analyzed. In the mice exposed to cyclophosphamide, reduction in the sperm count and viability and increase in the abnormal sperm and MDA levels were detected (p < .05). In addition, an increase in sperms with damaged DNA was detected in CP group, while the use of Ceratonia siliqua L. Extract significantly recovered these disturbances in the treatment group (p < .05). This study suggested the competence of Ceratonia siliqua L. extract in the improvement of sperm parameters and DNA fragmentation in animals treated with CP.
... The deleterious effects of CP have been well documented, and some agents, such as Phyllanthus fraternus Webster, have been used to protect the testicular structure of CP-treated mouse testes [15]. One study reported the protective effects of ginger on the sperm characteristics and epididymal morphology in rats treated with CP [16]. However, this study tested the combined effects of ginger and pumpkin seed extracts, and it is not clear if the protective effects were greater because of the ginger extract or the pumpkin seed extract. ...
Full-text available
Background: Cyclophosphamide (CP) is a widely used medication in chemotherapy and can cause oxidative stress. Oxidative stress can affect testicular functions by reducing the sperm motility and concentration, changing the sperm morphology, and increasing DNA fragmentation in sperm. Ginger is one of the most widely used spices in various foods and is used as an herbal medicine in many countries due to its antioxidant effects. We aim to study the protective effects of ginger against CP-induced testicular toxicity in rats. Objectives: This study was conducted to investigate the role of ginger in preventing cyclophosphamide-induced adverse effects on the testicular histology of CP-treated male rats. Methods: The study was performed on 30 male albino rats with body weights of 300-350 g. The animals were divided into the following three groups (10/cage): Group 1 (control, untreated group), Group 2 (CP group, received a single dose of CP at 100 mg/kg(-1) BW infraperitoneally), and Group 3 (CP+ginger, received ginger extract orally at 500 mg/kg for 35 days after CP injection). The morphological and histological structures of the testes were compared in the different groups of rats. Results: The CP-treated group showed a disorganized germinal epithelium compared with those of the controls. The CP+ginger-treated group showed a significant recovery of the organization of the germinal epithelium and the cellular attachments. Caspase-3-positive cells were significantly higher in the CP group and had remarkably lower levels in the CP+ginger-treated group. A reduction in the diameter of the seminiferous tubules and the destruction of connective tissue were observed in the CP-treated group; these changes were improved in the CP+ ginger-treated group. Conclusion: Ginger extract can protect reproductive functions against CP-induced toxicity in rats.
Cyclophosphamide is an alkylating agent that is widely used in cancer therapy. An important complication of this drug is hepatotoxicity. The hepatic prophylactic effects of ginger and pumpkin seeds were observed due to antioxidant effect. In the current study, the effects of ginger, pumpkin seed extracts, and their combination on cyclophosphamide-induced hepatotoxicity in rats were investigated. In this intervention study, a total of 70 male rats were randomly divided into 10 groups. The blood was taken after 2 weeks of treatment. Liver enzyme levels including ALT, AST and MDA were measured. Sections of the livers of all groups were provided and then histologically evaluated. The data obtained were analyzed using SPSS software version 16. A significant increase in liver enzymes was observed in rats treated with cyclophosphamide compared to other groups. Liver enzyme levels were decreased in rats treated with ginger, pumpkin seed and combined extracts (P < 0.05). Histological examination of liver samples treated with ginger, pumpkin seeds and combined extracts showed a significant reduction in liver damage compared to the cyclophosphamide-treated group (P < 0.05). Ginger, pumpkin seeds and combined extracts could improve biochemical parameters such as AST and ALT and pathological damage in the liver of cyclophosphamide-treated rats.
Full-text available
The inability to have a child is a baleful event for millions of couples in their life, and a large percentage of them have a personal frustration. The problem of infertility in couples is distributed equitably between the two sexes. Among different methods, medicinal plants have been used in many Nations to treat male infertility problems. These medicinal herbs are used to treat sperm disorders, dysfunctioning of the libido, 2 sexual asthenia and erection. Herbs provide a therapeutic option, which is affordable and available for infertile couples, and herbalism is the main form of treatment in our health system. So in this review, we have summarized most of the data dealing with the positive effects of plant extracts on mammalian reproductive system.
Full-text available
To investigate the effects of Zingiber Officinale on male reproductive functions and study the mechanisms underlying these effects, aqueous extract of Zingiber Officinale were administered orally to two groups of male rats at 500mg/kg b.w. and 1000mg/kg b.w. A third group served as control and received the treatment vehicle, distilled water. Treatment lasted for 14 and 28 days before sacrifice. Organ weight, epididymal sperm counts, motility, viability and morphology, seminal fructose, testicular malonhydialdehyde, and serum testosterone were determined. The treatment caused a significant increase (P<0.05) in the weight of the testis and epididymis. There were dose and duration dependent increases in sperm count and motility (P<0.05). There was also a significant increase (P<0.05) in serum testosterone level. Malonhydialdehyde levels were significantly reduced (P<0.05). Our results indicated that extract of Zingiber Officinale possesses pro-fertility properties in male rats which might be a product of both its potent antioxidant properties and androgenic activities. (Afr. J. Biomed. Res. 11: 329 - 334 ) Key Words: Zingiber Officinale, testosterone, androgenic, malonhydialdehyde and sperm.
Full-text available
Carbendazim is a carbamate fungicide used in control of various fungal pathogens.Fenugreek (Trigonella foenum-graecum Linn.) is a leguminous plant cultivated in several Asian and African countries and its seeds are used as herbal medicine. In the present work the effect of aqueous extract of fenugreek seeds on carbendazim-induced testicular toxicity in albino rats was studied. Treating rats with carbendazim induced significant decrease in testis weights, diameters and germinal epithelial heights of the seminiferous tubules. Histological results revealed degeneration of seminiferous tubules and reduction of spermatogenic cells. Moreover, carbendazim caused elevation of testicular malondialdehyde (MDA), and reduced the activity of the antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT). Animals treated with carbendazim and fenugreek showed improvement in the histomorphological and histopathological changes observed in animals treated with carbendazim. In addition fenugreek treatment leads to a significant decrease in the level of MDA and increase in the activity of SOD and CAT. It is concluded that fenugreek extract can improve the testicular toxicity of carbendazim and this effect may be attributed to its antioxidant properties.
Objective: To investigate the comparative effects of aminoglycosides and fluoroquinolones on testis structure and serum testosterone hormone level in rats. Methodology: Forty male Wister rats were randomly divided into control (n=10) and experimental (n=30) groups. The experimental groups were subdivided into three groups of ten. Each received 5 mg/kg (IP) Gentamicin, 40mg/kg (IP) Streptomycin and 72mg/kg (IP) Ofloxacin daily for 14 days, respectively; however, the control group just received vehicle (IP). In the fourteenth day, 5cc blood was collected for testosterone hormone then rats were killed and testis tissues were also prepared for light and electron microscopic study. Results: Depletion of germ cells, germinal cells necrosis, especially in spermatogonia, and Leydig cells had an abnormal fibroblast-like appearance. Abnormal space between neighbour sertoli cells, mitochondria were lost cristae and vacuolated (none energized), lyzosome seen more in cytoplasm of sertoli cells and Veins congestion were seen in gentamicin & ofloxacin groups. These side effects were seen fewer in Streptomycin group. Conclusions: Gentamicin, Streptomycin and Ofloxacin have negative effects on testis architecture and germinal cells damages in rats. However, these side effects are seen less in the Streptomycin group. Therefore, it is recommended that usage of this drug have fewer side effects on male fertility.
Zerumbone (ZER), a sesquiterpene from the edible plant Zingiber zerumbet Smith, has recently been found to suppress tumor promoter 12- O -tetradecanoylphorbol-13-acetate (TPA)-induced Epstein–Barr virus activation in a potent manner. In the present study, we evaluated the anti-inflammatory and chemopreventive potentials of ZER in a variety of cell culture experiments. ZER effectively suppressed TPA-induced superoxide anion generation from both NADPH oxidase in dimethylsulfoxide-differentiated HL-60 human acute promyelocytic leukemia cells and xanthine oxidase in AS52 Chinese hamster ovary cells. The combined lipopolysaccharide- and interferon-γ-stimulated protein expressions of inducible nitric oxide synthase and cyclooxygenase (COX)-2, together with the release of tumor necrosis factor-α, in RAW 264.7 mouse macrophages were also markedly diminished. These suppressive events were accompanied with a combined decrease in the medium concentrations of nitrite and prostaglandin E 2 , while the expression level of COX-1 was unchanged. ZER inhibited the proliferation of human colonic adenocarcinoma cell lines (LS174T, LS180, COLO205, and COLO320DM) in a dose-dependent manner, while the growth of normal human dermal (2F0-C25) and colon (CCD-18 Co) fibroblasts was less affected. It also induced apoptosis in COLO205 cells, as detected by dysfunction of the mitochondria transmembrane, Annexin V-detected translocation of phosphatidylserine, and chromatin condensation. Intriguingly, α-humulene, a structural analog lacking only the carbonyl group in ZER, was virtually inactive in all experiments conducted, indicating that the α,β-unsaturated carbonyl group in ZER may play some pivotal roles in interactions with unidentified target molecule(s). Taken together, our results indicate that ZER is a food phytochemical that has distinct potentials for use in anti-inflammation, chemoprevention, and chemotherapy strategies.
Spermatozoa leaving the testis contain a cytoplasmic droplet which they release during transit through the epididymis before reaching the cauda epididymidis. The cytoplasmic droplet shows P450 aromatase activity, which plays a role in synthesis of oestrogen from androgen. In the present study, 3-month-old Wistar strain male albino rats were administered with the organophosphate insecticides malathion or dichlorvos, or the phytotherapeutics andrographolide or ursolic acid. Segments of the epididymis were subjected to histopathological and ultrastructural analyses and it was found that 60-95% of the spermatozoa residing in the lumen of the cauda epididymidis retained the cytoplasmic droplet. The motility of the spermatozoa released from the cauda epididymidis was inhibited. One of the mechanisms of action of these toxicants on male reproductive function may be attributed to the retention of the cytoplasmic droplet and the resultant impairment of sperm motility.
The poor bioavailability and stability of curcumin limit its clinical application. A novel Zn(II)-curcumin complex was synthesized and its effects against cyclophosphamide (CP)-induced reproductive damage were compared with curcumin. Oral administration of Zn(II)-curcumin significantly prevented CP-induced elevation of malondialdehyde (MDA) level and reductions in superoxide dismutase (SOD) activity and glutathione (GSH) content in mouse testis. Zn(II)-curcumin significantly ameliorated CP-induced reductions in body and reproductive organs weights. Zn(II)-curcumin dose-dependently ameliorated CP-induced reproductive system impairments, by improving sperm parameters (sperm count, viability, motility) and reducing serum testosterone and histological alterations. Compared to curcumin at the same dose, Zn(II)-curcumin more effectively alleviated CP-induced reproductive injury, leading to a reduced severity of testicular pathologic changes, lower MDA level, elevated SOD activity and GSH content, and increased sperm parameters and serum testosterone. These results suggest Zn(II)-curcumin more effectively protects against CP-induced reproductive damage than curcumin alone due to a synergistic reduction in oxidative stress.
Treatment with cyclophosphamide, a commonly used anticancer drug, may result in oligozoospermia or azoospermia. The objective of this study was to determine whether exposure of male rats to cyclophosphamide induces apoptosis in male germ cells, and if so, when the peak of apoptosis occurs and at what specific stages of spermatogenesis. The presence of apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) detection in situ and by an increase in DNA fragmentation (DNA ladder). To determine the time course of drug-induced apoptosis, male Sprague-Dawley rats were treated with a single dose (70 mg/kg BW) of cyclophosphamide, and the testes were fixed 0, 4, 8, 12, 18, 24, and 48 h after treatment. To determine the dose response, rats were treated with doses of cyclophosphamide (0, 2, 7, 20, and 70 mg/kg), and the testes were fixed 12 h after treatment. A low spontaneous incidence of apoptosis was observed in controls, in particular in premeiotic germ cells of stages I-IV and XI-XIV of the seminiferous tubules. In cyclophosphamide-exposed rats, the incidence of apoptosis increased progressively at 4 h and 8 h, reached a peak at 12 h (about 3.5-fold above control), and then decreased rapidly to control levels by 48 h. A 70-mg/kg dose of cyclophosphamide induced a significant increase in apoptosis; lower doses did not. Although drug-induced apoptosis occurred in all stages of germ cells, it was most pronounced in spermatogonia and spermatocytes in stages I-IV and XI-XIV. Thus, apoptosis may be involved in the occurrence of oligozoospermia or azoospermia after cyclophosphamide treatment. Apoptosis of damaged premeiotic germ cells may serve a critical role in protecting subsequent generations from the diverse effects of toxicants.