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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats The 5 th Arab and 2 nd International Annual Scientific Conference on: Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education Institutions in Egypt and

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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats *Abd El-Ghany, M. A ** Dalia, A. Hafez ** Soha, M. Sameh El-Safty* Abstract The present study was designed to investigate some chemical composition of pumpkin seeds and also determine the effect of pumpkin seeds and / or zinc on the nutritional and sexual healthy status. Forty-nine adult albino male rats Sprague –Dawley strain were classified into control negative (-ve) group and six rat groups which administered 50 mg of lead/kg body weight per day as testicular toxicant and classified into control positive (+ve), pumpkin extract, pumpkin oil, zinc, pumpkin extract with zinc and pumpkin oil with zinc groups. The study was assigned for three month. The results revealed that, the control (+ve) group showed a significant decrease in weight gain; feed efficiency ratio; serum LH, FSH& testosterone; blood GPX & SOD and testis SOD, catalase, GPX, zinc& copper but a significant increase in testis LPX compared with control (- ve) group. The pumpkin extract rat group showed a significant decrease in weight gain; feed efficiency ratio; blood GPX & SOD and testis copper while the pumpkin oil rat group showed a significant decrease in weight gain; feed efficiency ratio ,blood GPX & SOD and testis copper but significant increase in testis LPX compared with control (- ve) group. Zinc rat group showed a significant decrease in weight gain ,feed efficiency ratio and blood GPX & SOD but significant increase in testis LPX while the pumpkin extract with zinc and pumpkin oil with zinc groups showed a significant decrease in weight gain and feed efficiency ratio but a significant increase in serum LH compared with control (- ve) group. The histopathological results showed that pumpkin extract, pumpkin oil and zinc rat groups had lower changes of testis but pumpkin extract with zinc and pumpkin oil with zinc rat groups showed normal structure of testis. In conclusion, the administration of pumpkin seeds and zinc can lower the side effects of a lead testicular toxicant and improve the healthy status of testis with increase of reproductive potential. Pumpkin seeds can be considered as one of aphrodisiacs.
Biological study on the effect of pumpkin
seeds and zinc on reproductive potential
of male rats
* Abd El-Ghany, M. A
** Dalia, A. Hafez
** Soha, M. Sameh El-Safty
* Home Economics Dept, Faculty of Specific Education, Mansoura University, Egypt.
**Home Economics Dept, Faculty of Education, Suez University, Egypt.
The 5
th
Arab and 2
nd
International
Annual Scientific Conference on:
Recent Trends of Developing Institutional and
Academic Performance in Higher Specific Education
Institutions in Egypt and Arab World
Faculty of Specific Education
Mansoura University - Egypt
April, 14-15, 2010
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2384
Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
2385
Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
Biological study on the effect of pumpkin seeds
and zinc on reproductive potential of male rats
*Abd El-Ghany, M. A
** Dalia, A. Hafez ** Soha, M. Sameh El-Safty
*
Abstract
The present study was designed to investigate some chemical
composition of pumpkin seeds and also determine the effect of pumpkin seeds
and / or zinc on the nutritional and sexual healthy status. Forty-nine adult albino
male rats Sprague –Dawley strain were classified into control negative (-ve)
group and six rat groups which administered 50 mg of lead/kg body weight per
day as testicular toxicant and classified into control positive (+ve), pumpkin
extract, pumpkin oil, zinc, pumpkin extract with zinc and pumpkin oil with zinc
groups. The study was assigned for three month.
The results revealed that, the control (+ve) group showed a significant
decrease in weight gain; feed efficiency ratio; serum LH, FSH& testosterone;
blood GPX & SOD and testis SOD, catalase, GPX, zinc& copper but a
significant increase in testis LPX compared with control (- ve) group. The
pumpkin extract rat group showed a significant decrease in weight gain; feed
efficiency ratio; blood GPX & SOD and testis copper while the pumpkin oil rat
group showed a significant decrease in weight gain; feed efficiency ratio ,blood
GPX & SOD and testis copper but significant increase in testis LPX compared
with control (- ve) group.
Zinc rat group showed a significant decrease in weight gain ,feed
efficiency ratio and blood GPX & SOD but significant increase in testis LPX
while the pumpkin extract with zinc and pumpkin oil with zinc groups showed
a significant decrease in weight gain and feed efficiency ratio but a significant
increase in serum LH compared with control (- ve) group.
The histopathological results showed that pumpkin extract, pumpkin oil
and zinc rat groups had lower changes of testis but pumpkin extract with zinc
and pumpkin oil with zinc rat groups showed normal structure of testis.
In conclusion, the administration of pumpkin seeds and zinc can lower
the side effects of a lead testicular toxicant and improve the healthy status of
testis with increase of reproductive potential. Pumpkin seeds can be considered
as one of aphrodisiacs.
* Home Economics Dept, Faculty of Specific Education, Mansoura University, Egypt.
**Home Economics Dept, Faculty of Education, Suez University, Egypt.
2386
Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
Biological study on the effect of pumpkin seeds
and zinc on reproductive potential of male rats
*
Abd El-Ghany, M. A
** Dalia, A. Hafez ** Soha, M. Sameh El-Safty
*
INTRODUCTION
Reproductive ability in the male comprises the production of semen
containing normal spermatozoa in the adequate number, together with the desire
and ability to mate (Oyeyemi et al., 2008). For many years, people have
searched for ways to achieve sexual desire, sexual health and sexual techniques
as aphrodisiacs. An aphrodisiac may be food, drug, scent or device that
enhances sex drive and/or sexual pleasure (Rosen and Ashton, 1993). The use
of alternative medicine has increased considerably in the last 10 years. Some of
the reasons for this trend include limited efficacy of existing treatments,
perceived reduction in side effects with alternative treatments, patient's desire to
maintain control over their treatment and a desire for a more "natural" treatment
(Shoskes, 2002).
Lead is a common environmental and industrial pollutant that is
detected in many phases of environment and biological systems. The most
important sources of lead exposure are industrial emissions, soils, car exhaust
gases and contaminated foods. Vegetables with a relatively large leaf area, such
as spinach and cabbage can contain high levels when grown near lead sources
(Gama et al., 2006). Lead is known to disrupt the pro-oxidant/antioxidant
balance of tissues, which leads to biochemical and physiological dysfunction.
Exposure to lead causes various toxicological effects as a result of their
distribution in various tissues and organs, including the stomach, intestines,
kidney, liver, spleen, heart, bone, and nervous and reproductive systems
(Adnan and AL-Safi, 2005). Lead is well documented as a testicular toxicant
and has been shown to perturb reproductive capability. Lead may inhibit
spermatogenesis and decrease in the young spermatids; pachytene
spermatocytes and mature spermatids (Kaushal et al., 1996).
Pumpkin (Cucurbita pepo L.) seed and seed oil have macro- and micro-
constituent composition. They are a rich natural source of proteins, phytosterols
* Home Economics Dept, Faculty of Specific Education, Mansoura University, Egypt.
**Home Economics Dept, Faculty of Education, Suez University, Egypt.
2387
Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
polyunsaturated fatty acids, antioxidant vitamins, such as carotenoids and
tocopherol and trace elements, such as zinc. Pumpkin seed extract contain high
levels of natural potent phytochemicals sterols with great promise for
immunomodulation, reproductive health, and therapeutic advantage over a wide
range of disease conditions. The intake of a whole extract of pumpkin seeds is
correlated to reduced benign prostate hyperplasia-associated symptoms (Glew
et al., 2006, Fruhwirth and Hermetter, 2007 and Stevenson et al., 2007).
Pumpkin seeds have been used for many years to improve sexual stimulation
and improvement of sexual performance in terms of intromissions and
ejaculatory latency which also improved sexual sensation and coupulatory
efficiency (Gundidza et al., 2009).
Zinc is an essential mineral, important in prostate gland function and the
growth of the reproductive organs. Zinc is found more in male reproductive
fluid than anywhere else in the body. It is required for protein synthesis and
collagen formation, promotes a healthy immune system, the healing of wounds
and synthesis of DNA and RNA. Zinc is an important fertility nutrient for both
sexes. Zinc deficiency is also linked to men’s infertility. Zinc will normalize
deficient sperm counts and sperm motility. Even marginal zinc deficiency can
cause sperm counts to drop below the point of technical sterility (Feng et al.,
2002).
Therefore, the present study was designed to investigate some chemical
composition of pumpkin seeds and also determine the effect of pumpkin seeds
and / or zinc on the nutritional and sexual healthy status.
MATERIALS AND METHODS
A – Materials
1- Lead acetate, zinc and pumpkin seed oil:
Lead acetate is a white powder odorless, with a chemical structure
(CH
3
COO) 2Pb
3
H
2
O and was purchased from El-Gomhoria Co., El-Mansoura
city, Egypt. The dose of lead acetate was 50 mg of lead/kg body weight per day
according to previous studies as that recorded by Batra et al., (2004). Zinc was
obtained from October Pharma Co., October city, Egypt as Octazone capsules.
Each capsule contains 110 mg zinc sulphate equivalent to 25 mg zinc .The
human therapeutic dose of zinc was 25 mg Zn daily which converted to animal
dose (4.5 mg/kg) according to Paget and Barnes, (1964). Pumpkin seed oil
was obtained from Arab Company for Pharmaceutical and Medicinal Plants,
MEPACO'',Egypt . The human therapeutic dose was 320 mg /day. The
equivalent rat dose was of 288 mg/kg b.wt (Hongji and Jongro-gu, 2009)
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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
2- Pumpkin seeds:
Pumpkin fruits were obtained from a private farm in Minufia
governorate. Pumpkin seeds called pepitas, are the flat, green or white seeds
found inside pumpkins and obtained manually .The outer core of the pumpkin
seed was removed by hand milling and sieving. Pumpkin seeds were subjected
to dry freezing, then crushed to powder for ethanol extraction.
3 -Experimental animals:
Forty-nine adult albino male rats Sprague –Dawley strain were purchased
from Helwan Farm of Laboratory Animals. The average weight was 154 ±7 g.
The animals were kept under observation for five days before experiment and
fed on standard diet according to NRC, (1995) and water ad libitum. The
standard diet comprised of casein (200g/kg), corn starch (497g/kg), sucrose
(100g/kg), cellulose (30 g/kg), corn oil (50g/kg), mineral mixture (100g/kg),
vitamins mixture (20g/kg) and DL-methionine (3g/kg).
B- Methods:
1-Determination of the gross chemical composition and minerals of
pumpkin seeds:
Protein, fat, ash, and moisture and some minerals as zinc, iron calcium,
magnesium and sodium of pumpkin seeds were determined according to the
methods of the A.O.A.C, (2000). The total carbohydrates were calculated as
following:
Carbohydrates % = 100 - (moisture % + protein % + fat % + ash %).
2- Preparation of the pumpkin seeds 80% ethanolic extract:
100 g of air-dried, powdered pumpkin seeds were soaked in 500 ml of
80% ethanol with frequent agitation. Clarification was then carried out using
vacuum filtration through filter paper watman number 2. The resultant extract
was concentrated to dryness in a rotary evaporator under reduced pressure at a
temperature of 40°C. The rate dose of pumpkin seeds extract was 40 mg/kg
b.wt according to Marianna et al., (2009).
3- Grouping of rats and experimental design:
The rats were randomly classified into seven groups (7 rats each) and
fed on the standard diet. The rats classified into control negative (–ve) group
and six rat groups which administered lead acetate and reclassified into
untreated control positive (+ve),and treated groups which were pumpkin
extract, pumpkin oil, zinc, pumpkin extract with zinc and pumpkin oil with zinc
groups.
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Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
All treatments were given in a 1ml volume from stock solution by
stomach tube all over the period of the experiment. The study was assigned for
three month. The food intake was calculated daily and the body weight gain
was recorded weekly. Feed efficiency ratio was determined according to the
method of Chapman et al., (1950).
4- Collection of blood and testis samples:
At the end of the experiment, the rats were sacrificed to obtain blood
samples. Part of blood was heparenized while the rest part of blood was left to
coagulate then centrifuged at 3000 rpm for 15 minutes to obtain serum for each
individual sample and then stored at -20 °C for some laboratory analyses. Testis
of male rats was rapidly removed. The right testis was homogenized with 15
volumes of 0.58% K CL solution at 4
o
C. The homogenate was centrifuged at
1000g for 10 min. The supernatant was used for the assay of some laboratory
analyses.
5- Determination of some blood and testis biochemical parameters:
Blood glutathione peroxidase (GPX) and superoxide dismutase (SOD)
were estimated according to Beuther et al., (1987) and Beuchamp and
Fridovich, (1971), respectively. Serum luteinizing hormone, follicle
stimulating hormone and testosterone hormone were estimated according to
Loraine and and Bell, (1976), McCann and Kirkish, (1985) and Bee and
Kah, (2003), respectively. Testis superoxide dismutase (SOD), catalase,
glutathione peroxidase (GPX) and lipidperoxide (LPX) were estimated
according to Kono, (1978), Aebi, (1974), Lawrence and Burk, (1976) and
Placer et al., (1966), respectively. Testis zinc and copper were also estimated
according to Versiek, (1984).
6-Histopathological examination of the testis:
The left testis was fixed in 10% neutral buffered formaldehyde solution
at pH 7.5 and cleared in xylol and embedded in paraffin. 4-5 µm thick section
were prepared and stained with Hematoxylin and Eosin (H&E) for subsequent
histopathological examination according to Bancroft et al, (1996).
C-Statistical analysis
All the obtained data were statistically analyzed by SPSS computer soft
ware. The calculated occurred by analysis of variance ANOVA and follow up
test LSD by SPSS ver.11 according to Armitage and Berry (1987).
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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
RESULTS
1- The chemical composition of pumpkin seeds results:
The statistical date in table (1) denoted that the percentage values of
protein, fat, ash, fiber, moisture, and carbohydrate (CHO) in pumpkin seeds
were 31.57, 29.01, 3.89, 6.36, 5.11 and 24.06 %. The values of zinc, iron,
calcium, magnesium and sodium were 7.99, 9.76, 78.18, 90.69 and 20.56
mg/100g.
2-Nutritional results:
Data in table (2) illustrated that, the control (+ ve) group had a
significant lower values of the body weight gain and feed efficiency ratio
(p<0.01) compared to control (- ve) group. The pumpkin extract, pumpkin oil,
zinc, pumpkin extract with zinc and pumpkin oil with zinc groups showed a
significant lower values of the body weight gain and feed efficiency ratio
(p<0.05) but a non significant difference in final body weight and food intake
compared to control (- ve) group.
Analysis of variance ANOVA and LSD tests revealed that , the pumpkin
extract, pumpkin oil, zinc, pumpkin extract with zinc and pumpkin oil with zinc
groups showed a significant higher values of the body weight gain and feed
efficiency ratio but a non significant difference in final body weight and food
intake compared to control (+ ve) group
3-Biochemical results:
The results in table (3) recorded that control (+ ve) group showed a
significant decrease in serum luteinizing hormone, follicle stimulating hormone
and testosterone hormone (p< 0.05 & 0.001) compared to control (- ve) group.
The pumpkin extract, pumpkin oil, zinc, and pumpkin extract with zinc groups
showed non significant difference in serum luteinizing hormone, follicle
stimulating hormone and testosterone hormone (p>0.05) but pumpkin oil with
zinc group showed a significant increase in serum luteinizing hormone(p<0.05)
and non significant difference in serum follicle stimulating hormone and
testosterone compared to control (-ve) group.
Analysis of variance ANOVA and LSD tests revealed a significant
higher values of serum luteinizing hormone, follicle stimulating hormone and
testosterone hormone in the pumpkin extract, pumpkin oil, zinc, pumpkin
extract with zinc and pumpkin oil with zinc groups compared to control (+ ve)
group.
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Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
Data in table (4) illustrated that, the control (+ ve) group showed a
significant lower values of blood glutathione peroxidase and superoxide
dismutase (p<0.01&0.001) compared to control (- ve) group. The pumpkin
extract, pumpkin oil and zinc groups also showed a significant lower values
(p<0.05&0.01) but pumpkin extract with zinc and pumpkin oil with zinc groups
showed non significant differenc in blood glutathione peroxidase and
superoxide dismutase compared to control (- ve) .
Analysis of variance ANOVA and LSD tests revealed a significant
higher values of blood glutathione peroxidase and superoxide dismutase in
compared to control (+ ve) group.
The results in table (5) recorded that control (+ ve) group showed a
significant decrease in testis superoxid dismutase, catalase and glutathione
peroxidase and significant increase in lipid peroxide (LPX) (p<0.001) but the
pumpkin extract, pumpkin oil, zinc, pumpkin extract with zinc and pumpkin oil
with zinc groups showed non significant difference in superoxid dismutase,
catalase and glutathione peroxidase compared to control (- ve) group. The
pumpkin oil and zinc groups showed a significant increase in lipid peroxide
(p<0.05) compared to control (- ve) group.
Analysis of variance ANOVA and LSD tests revealed significant higher
values of testis superoxid dismutase, catalase and glutathione peroxidase and
significant decrease in lipid peroxide in pumpkin extract, pumpkin oil, zinc,
pumpkin extract with zinc and pumpkin oil with zinc groups compared to
control (+ve) group.
Data in table (6) illustrated that, the control (+ ve) group showed a
significant lower values of testis zinc and copper (p<0.01&0.001) but the
pumpkin extract and pumpkin oil groups showed a significant lower values of
testis copper only (p<0.05) compared to control (- ve) group. The zinc,
pumpkin extract with zinc and pumpkin oil with zinc groups showed non
significant difference in values of testis zinc and copper compared to control (-
ve) group.
Analysis of variance ANOVA and LSD tests revealed significant higher
values of testis zinc and copper of the pumpkin extract, pumpkin oil, and zinc,
pumpkin extract with zinc and pumpkin oil with zinc groups compared to
control (+ve) group.
4-Histopathological results:
The obtained results are confirmed by the histopathological changes.
Macroscopically examination of the testis of rat from the control (- ve) group
revealed normal semineferous tubules (Pict.1). Meanwhile, the testis of rat from
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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
the control (+ve) group revealed orchitis with infiltration of intraluminal
leucocytic cells and spermatide giant cells normal semineferous tubules (Pict.2).
Testis of rat from pumpkin extract group showed slight degeneration of
spermatogoneal cells (Pict.3).The testis of rats from pumpkin oil showed mild
interstitial edema (Pict. 4). The testis of rats from zinc group showed slight
degeneration of spermatogoneal cells lining seminiferous tubules (Pict. 5). The
testis of rat from pumpkin extract with zinc and pumpkin oil with zinc groups
showed normal structure of seminefrous tubules (Pict. 6 and 7).
DISCUSSION
The obtained results of the chemical composition of seeds were varied
than the previous results that may be due to different cultivars. Aboul-Nasr et
al., (1997) recorded that the chemical composition of pumpkin seeds were
40.27 % protein, 34.59% crude oil, 13.79% crude fiber and 4.45% ash.
Bombardelli and Morazzoni, (1997) and Nakiae, et al., (2006) found that the
nutritional value of pumpkin seeds is based on high protein content (25–51%)
and high percentage of oil that ranging from 40% to 60%, up to 60.8%, of the
oil is from the fatty acids oleic (up to 46.9%), linolenic (up to 40.5%), palmitic
and stearic up to 17.4%.There are approximately 4–5% minerals including
selenium, zinc, calcium, copper, iron, manganese, phosphorous and potassium;
approximately 30% pectin .
The occupational or experimental exposure to lead is associated with
increased oxidative reaction, which might be responsible, at least in part, for
lead-induced toxic effects. The heavy metals not only affect the nutritive values
of fruits and vegetables but also have deleterious effect on consumer of these
food items. Lead can depress the growth which is associated with the dose and
duration of lead (Lawton and Donaldson, 1991). Lead exposure causes a
significant increase in lipid peroxidation due to the high affinity of lead for
sulfhydryl groups or metal cofactors in these enzymes and substrate molecules.
The increase in the lipid peroxidation is accompanying with inhibition in the
antioxidant enzymes activities such as SOD and catalase because lead induces
copper deficiency and copper plays a catalytic role in this enzyme. Catalase also
works in close association of SOD. Lead interferes with absorption of iron
resulting in inhibition of hem biosynthesis that decreases in catalase activity.
Lead also can change in the concentrations and distribution of essential
elements in an organism as copper and zinc (Gasiorowski et al., 1987 and
Patra and Swarup ,2000). Lead exposure shifts in the pro-oxidant/antioxidant
balance resulting in increased free-radical generation, lipid peroxidation, and,
consequently, oxidant stress (Adonalyo and Oteiza, 1999 and Patra et al.,
2001).The presence of free radicals in various organs including the testis is a
2393
Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
normal physiological event; however, the alterations in their synthesis stimulate
the oxidation and DNA damage of cells. The plasma membrane of sperms
contains a high amount of unsaturated fatty acids so it is particularly susceptible
to peroxidative damage. The lipid peroxidation destroys the structure of the
lipid matrix in the membranes of spermatozoa, and it is associated with loss of
motility and the defects of membrane integrity. Exposure to stress causes a fall
of the testosterone level in peripheral blood of man and various species of
animals. Changes in the testosterone concentration are associated with
depression of the secretory activity of the testes, and changes in peripheral
metabolism (Katsiya et al., 1989 and Abd El-Ghany, 2007). Luteinizing
hormone is a hormone produced in the anterior pituitary gland, which
stimulates the release of sex hormones by the ovaries and testes. In men it
induces secretion of testosterone by the interstitial cells of the testes. The
reduction in LH and FSH in lead exposure due to adverse effect on
hypothalamic pituitary testicular hormonal axis and also impair of a negative
feedback control of testosterone by pituitary LH synthesis and or of
hypothalamic LH releasing hormone secretion (Gorbel et al .,2002). The
reduction in FSH in lead exposure could be due to impairment of some sertoli
cell protein secretion (Rotten, 1991).
Glutathione, glutathione peroxides, catalase, superoxide-dismutase are
biological antioxidants, which suppress or scavenge of free radical that is likely
to improve sperm function (Sanocka and Kurpisz, 2004 and Henkel, 2005).
Pumpkin seeds promote prostate and urological health and help to reverse the
age-related decline in the health of the prostate gland. Pumpkin seeds are also
an excellent source of magnesium, phosphorus, manganese, copper and iron in
healthy amounts in addition to zinc which is a nutrient vital to healthy
functioning of the male reproductive system (Alan, 2006). Pumpkin seeds are
rich in unsaturated fatty acids such as omega 3, 6 and 9 as well as its high
protein therefore the rats which fed diets rich in monounsaturated fats had
greater –dehydrogenase activity which is a key enzyme in the testosterone
synthesis pathway in the male rat and plasma androgen concentrations
compared to rats fed diets rich in saturated and polyunsaturated fats (Nagata, et
al., 2000). Pumpkin seed inhibit the conversion of testosterone into
dihydrotestosterone in cultures of human fibroblasts. Men's health products
commonly contain pumpkin seeds because pumpkin seeds are a rich source of
zinc which is thought to promote prostate health and to improve bone mineral
density (Bach, 2000 and Liu et al., 2007). The elevated levels of serum
testosterone may be one of the mechanisms underlying the effect of squalene in
pumpkin seeds on improvement in libido and semen quality and the reduction
in serum leptin (Banks, et al., 2004 and Bjorbaek and Kahn, 2004).
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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
Pumpkin seed oil is rich in many potent antioxidants and beneficial
nutritional supplements such as essential fatty acids including linoleic and
linolenic, ,carotenes, lutein, gamma and P-tocopherols, phytosterols,
chlorophyll , selenium and zinc, (Lee, 2006 and Gossell-Williams et al.,
2006). Essential fatty acids are required constituents of every membrane in the
body, playing a crucial role in maintaining the health of every living cell in the
body. They maintain the fluidity of cellular membranes, aid in producing and
balancing hormones, and play an essential role in managing healthy fluid levels.
The body metabolizes these fatty acids into a group of components known as
prostaglandins. Prostaglandins regulate every organ system in the body.
Pumpkin seed oil is reported to protect older men’s urinary tracts from the
aging effects of testosterone (Jayaprakasam, et al., 2003 and Fu et al., 2006).
The testosterone, LH and FSH concentration were close to normal with
treatment with pumpkin extract, oil and zinc due to all the steroid hormone
receptors require zinc ion to maintain their secondary structure and function.
Availability of more zinc might reactivate the normal functioning of receptors
(Bataineh et al., 2002).
Zinc is found in high concentrations in the human prostate gland for a
normal function. Zinc plays an important role in the structure of proteins and
cell membranes and protect against damage so plays important roles in growth
and development of the immune response, neurological function and
reproduction (Bataineh et al., 2002 and Vartsky et al., 2003). A deficiency in
zinc, blocks the essential rate-limiting enzyme Delta- 6-desaturase in the
transformation of essential fatty acids into the important prostaglandins. Zinc is
a cofactor for proper functioning, as carbonic anhydrase, alkaline phosphatase,
and superoxide dismutase. Loss of zinc from biological membranes increases
their susceptibility to oxidative damage and impairs their function. Zinc acts as
an antioxidant by protection of proteins and enzymes against free radical attack
or oxidation and prevention of free radical formation. The enzymes involved in
testosterone synthesis rely on zinc supplements could stimulate testosterone
production and consequently the number of sperm, (Takeda et al., 2005 and
Osaretin and Gabriel 2008).
The results of this study refocused on the importance of pumpkin seeds
either extract or oil and or zinc in improving reproductive potential of male rats.
So, it is advised to consume pumpkin seeds and zinc rich diet daily to decrease
the undesirable side effect of lead contaminants and improve the sexual health
status.
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Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
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nd
International
Annual Scientific Conference
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Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
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Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
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International
Annual Scientific Conference
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2399
Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
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Table (1): Some major and minor composition in pumpkin seeds CHO%
Moisture%
Fiber%
Ash %
Fat% Protein%
24.06
5.11
6.36
3.89
29.01
31.57
Sodium
(mg/100g)
Magnesium
(mg/100g)
Calcium
(mg/100g)
Iron
(mg/100g)
Zinc
(mg/100g
)
20.56
90.69
78.18
9.76
7.99
Table (2): Mean values ± SD of body weight gain, food intake and feed efficiency ratio
(FER) of the experimental rats groups.
pumpkin
oil with
zinc
pumpkin
extract
with zinc
Zinc
pumpkin
oil
pumpkin
extract
Control
(+ve)
Control
(-ve)
Groups
Variable
154.75
±
6.32
a
152.77
±
6.14
a
153.25
±
6.11
a
154.01
±
5.31
a
153.71
±
5.81
a
153.51
±
5.22
a
152.41
±
5.17
a
Initial
weight
(g)
213.64
±
10.88
a
212.75
±
12.31
a
207.41
±
11.13
a
209.24
±
11.11
a
207.22
±
10.11
a
198.22
±
9.17
a
223.27
±
10.24
a
Final
weight
(g)
58.89
±
4.01
b*
59.98
±
3.21
b*
54.15
±
3.41
b*
55.23
±
3.71
b*
53.51
±
3.25
b*
44.71
±
2.17
c**
70.88
±
7.33
a
Weight
gain
(g)
17.58
±
1.33
a
18.32
±
1.24
a
18.33
±
1.32
a
17.98
±
1.51
a
18.46
±
1.61
a
17.99
±
1.41
a
18.88
±
1.22
a
Food
intake
(g/d) 0.054
±
0.003
b*
0.054
±
0.002
b*
0.049
±
0.002
b*
0.051
±
0.003
b*
0.048
±
0.001
b*
0.041
±
0.002
c**
0.062
±
0.001
a
FER
Significant with control (-ve) group * P<0.05 ** P<0.01 *** P<0.001
Mean values in each raw having different superscript (a, b, c) denote significant difference.
2400
Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
Table (3): Mean values ± SD of serum luteinizing hormone (LH), follicle stimulating
hormone (FSH) and testosterone hormone of the experimental rats groups.
pumpkin
oil with
zinc
pumpkin
extract
with zinc
Zinc
pumpkin
oil
pumpkin
extract
Control
(+ve)
Control
(-ve)
Groups
Variable
1.01
±
0.11
b*
0.92
±
0.03
a
0.81
±
0.04
a
0.89
±
0.01
a
0.85
±
0.02
a
0.44
±
0.03
c**
0.91
±
0.01
a
LH
(ng
/
ml)
6.96
±
1.34
a
6.01
±
1.21
a
4.99
±
1.03
a
5.91
±
1.20
a
5.71
±
1.01
a
4.01
±
1.10
b*
6.68
±
1.71
a
FSH
(ng
/
ml)
5.01
±
0.15
a
4.11
±
0.13
a
3.44
±
0.11
a
3.98
±
0.04
a
3.67
±
0.71
a
2.11
±
0.30
b**
4.65
±
0.81
a
Testo-
sterone
(ng
/
ml)
Significant with control (-ve) group * P<0.05 ** P<0.01 *** P<0.001
Mean values in each raw having different superscript (a, b, c) denote significant difference.
Table (4): Mean values ± SD of blood glutathione peroxidase (GPX) and superoxide
dismutase (SOD) of the experimental rats groups.
pumpkin
oil with
zinc
pumpkin
extract
with zinc
Zinc
pumpkin
oil
pumpkin
extract
Control
(+ve)
Control
(-ve)
Groups
Variable 6.11
±
1.13
a b
5.99
±
1.41
a b
5.61
±
1.22
b*
5.31
±
1.43
b*
5.18
±
1.34
b*
3.15
±
0.51
c***
7.13
±
1.51
a
GPX
(ng/ml)
18.32
±
3.11
a b
17.11
±
2.55
a b
15.53
±
3.41
b**
14.71
±
3.21
b**
14.20
±
4.11
b**
10.21
±
2.14
c**
21.51
±
3.22
a
SOD
(ng/ml)
Significant with control (-ve) group * P<0.05 ** P<0.01 *** P<0.001
Mean values in each raw having different superscript (a, b, c) denote significant difference.
2401
Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
Table (5): Mean values ± SD of testis superoxid dismutase(SOD), catalase, glutathione
peroxidase (GPX) and lipid peroxide (LPX) of the experimental rats groups.
pumpkin
oil with
zinc
pumpkin
extract
with zinc
Zinc
pumpkin
oil
pumpkin
extract
Control
(+ve)
Control
(-ve)
Groups
Variable
6.99
±
1.03
a
7.01
±
1.61
a
6.59
±
1.31
a
6.31
±
1.51
a
6.49
±
1.20
a
3.44
±
0.59
b***
7.52
±
1.41
a
SOD
(µ/mg
protein)
14.61
±
2.35
a
14.51
±
2.11
a
14.43
±
2.61
a
13.25
±
3.11
a
14.01
±
2.61
a
7.11
±
1.31
b***
16.81
±
1.99
a
Catalase
(µ/mg
protein)
2.14
±
0.31
a
2.23
±
0.40
a
1.99
±
0.11
a
2.11
±
0.22
a
2.01
±
0.31
a
0.65
±
0.01
b***
2.33
±
0.60
a
GPX
(nmol)
5.32
±
0.15
bc
5.41
±
0.99
bc
6.13
±
0.81
b*
6.61
±
0.40
b*
5.58
±
0.35
bc
13.51
±
2.17
a***
4.33
±
0.41
c
LPX
(nmol)
Significant with control (-ve) group * P<0.05 ** P<0.01 *** P<0.001
Mean values in each raw having different superscript (a, b, c) denote significant difference.
Table (6): Mean values ± SD of testis zinc and copper of the experimental rats groups.
pumpkin
oil with
zinc
pumpkin
extract
with zinc
Zinc
pumpkin
oil
pumpkin
extract
Control
(+ve)
Control
(-ve)
Groups
Variable
25.71
±
4.25
a
26.11
±
4.89
a
26.69
±
5.43
a
21.11
±
5.30
a
22.32
±
3.45
a
17.32
±
3.12
b**
25.69
±
4.21
a
Zinc
(µ/mg)
2.55
±
0.31
a b
2.43
±
0.14
a b
2.69
±
0.28
a b
2.01
±
0.25
b*
2.17
±
0.30
b*
1.63
±
0.11
c***
3.03
±
0.45
a
Copper
(µ/mg)
Significant with control (-ve) group * P<0.05 ** P<0.01 *** P<0.001
Mean values in each raw having different superscript (a, b, c) denote significant difference.
2402
Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
Histopathological Examination of Testis
2403
Recent Trends of Developing Institutional and Academic Performance in Higher Specific Education
Faculty of Specific Education
Mansoura University - Egypt April, 8-9, 2009
2404
Biological study on the effect of pumpkin seeds and zinc on reproductive potential of male rats
The 5
th
Arab and 2
nd
International
Annual Scientific Conference
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Gאא Jאא Jא 
GGאא Jא Jא Jא J
... Experimental animals that received only ethanolic seed extract of Cucurbita pepo (Pumpkin) throughout the duration of the experiments showed an increase in the number of sperm count, percentage of normal sperm cells as well as higher serum levels of testosterone when compared to control and neem fed rats and seen in their respective histological slides. These improvements in testicular parameters when compared with control agrees with findings of Minisy et al., (2017), Aghaei et al., (2014), andAbd El-Ghany et al., (2010). According to Abd El-Ghany et al., (2010), pumpkin seeds improve sexual health status. ...
... These improvements in testicular parameters when compared with control agrees with findings of Minisy et al., (2017), Aghaei et al., (2014), andAbd El-Ghany et al., (2010). According to Abd El-Ghany et al., (2010), pumpkin seeds improve sexual health status. The increase in sperm count, and percentage of normal sperm cells seen in pumpkin seed treated group when compared to control and neem leaf fed rats obtained in this current study could be as a result of high antioxidant contents as well as vitamins E and A, and Carotenoids present in pumpkin seeds that have the potentials of breaking down the oxidative chain reaction and play a very significant role in increasing the body's capacity to fight free radical-induced oxidative stress, and therefore improve the process of spermatogenesis (Mohammadi et al., 2013). ...
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Cucurbita pepo seeds are highly nutritional, containing a wide array of healthy compounds and free-radical scavenging antioxidants that have been found useful in promoting good health. The aim of this study was to investigate the effect of methanolic seed extract of Cucurbita pepo (pumpkin) on Azadirachta Indica (neem) leaf induced testicular damage in adult male Albino Wistar rats. Twenty adult male albino wistar rats weighing between 110-230g were used, and assigned into 5 groups (I-V) of 4 animals each. Group I served as control. Groups II received 500mg/kg of ethanolic seed extract Cucurbita pepo (ESECP) for 5 weeks. Group III received 500mg/kg of methanolic leaf extract of Azadirachta indica (MLEAI) for 2 weeks and then 500mg/kg of ESECP for 3 weeks. Group IV received a co-administration of 500mg/kg of MLEAI and 500mg/kg of ESECP for 5 weeks, while Group V received 500mg/kg of for 5 weeks. All extracts were administered orally, once a day. Twenty four hours after the last administration, the rats were anesthetised and blood samples and testes were obtained for further investigation. Results showed significant increases in serum testosterone levels, improvements in sperm parameters (count, motility, and morphology) and better testicular microarchitecture in ESECP treated rats when compared to rats treated with MLEAI alone. This reveals that ethanolic seed extract of Cucurbita pepo exhibits both protective and ameliorative activities against Azadirachta indica induced testicular damage in adult male albino wistar rats.
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In poly cystic ovary syndrome multiple cysts formation occurs in female ovaries. It is highly prevalent in developing countries like Pakistan and India. Its symptoms include irregular menstrual cycle, abnormal hair growth pattern, acne and mood swings. This disease is also associated with imbalance in androgen levels (which include testosterone and estrogen) and high insulin levels also called hyperinsulinemia. It also contributes in the development of type 2 diabetes, heart disease and cancers. Seed cycling is a modern method used in alternate medicine of treating PCOS by using different seeds in the different stages of menstrual cycle. In this review article we will find out how seed cycling is effective in the treatment of PCOS. Flax, sesame, pumpkin and sunflower seed combinations are being used at different stages of female menstrual cycle. Flaxseeds contain lignans, omega 3 fatty acids and fibre which help in relieving the symptoms of polycystic ovaries. Sesame and sunflower seeds contain high number of fatty acids and lignans which helps in balancing hormonal disturbances in female. Losing weight by having diet rich in fibre improve the level of cholesterol and it is also beneficial decreasing the level of insulin resistance, which if not treated can progressed into type 2 diabetes mellitus and its complications. Herbal medicine in Ayurveda has shown promising effects in relieving pre- menstrual symptoms, hormonal fluctuations and the co-morbidities.
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Four healthy bucks of the West African Dwarf breed aged between 24 and 30 months were used for this study. The bucks were first used as control and later as experimental animals upon being fed daily with oiled pumpkin plant for the period of six months. The objective of the study was to investigate the effect of the pumpkin plant on the morphology of the spermatozoa of the bucks. There were significant differences (p< 0.05) between the control and experimental values for both primary and secondary morphological spermatozoa abnormalities: the pyriform head has a control value of 6 (0. 42%) and post-feeding value of 0 (0%), the beat tail; 14(0.97%) and 2 (0.16%) for the control and post feeding values respectively. The curved mid piece: 17 (1.18%) and 1 (0.08%) for the control and post-feeding values respectively. The bent mid piece also differed significantly (p<0.05) between control value of 16 (1.11%) and post feeding value of 3 (0.23%). All through the stages of the study, there were significant reductions in the number of sperm cells with abnormalities consequent upon daily feeding of the animals with pumpkin plant. The plant is therefore recommended for both prevention and treatment of make infertility in man and animals.
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The aim of the present study was to determine the effects of formulations composed of Mondia whitei, Ekebergia capansis, aloe tincture (Aloe exelsa) and pumpkin seed (Cucurbita pepo) on sexual behaviour of inexperienced male rats. Male rats were treated orally with ethanol extracts of M. whitei and E. capensis, aloe tincture (from aloe juice) and pumpkin seed powder. The sexual behaviour of the rats treated with extracts was compared with those treated with 100 µg of testosterone as the positive control and those receiving normal rat feed as negative control. The receptivity of the females was rendered homogeneous with daily dose of estadiol benzoate. The sexual behaviour of the rats were observed over one ejaculatory phase for five consecutive days. The results showed a dose dependent increase in sexual arousability (decreased mounting latency), copulatory efficiency (decreased intercoupulatory interval) and improved sexual sensation (increased in neuromotor activity) for the rats treated with M. whitei and E. capensis, aloe and pumpkin seeds showing an increase in sexual performance in terms of intromissions and ejaculatory latency which also improved sexual sensation and coupulatory efficiency. The formulations of plants under investigation showed significant aphrodisiac properties.
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Cucurbita pepo subsp. pepo var. Styriaca is a phylogenetically young member of the Cucurbita spp. since the mutation leading to dark green seeds with stunted outer hulls arose only in the 19th century. This mutation defined the so-called Styrian oil pumpkin and facilitated the production of Styrian pumpkin seed oil. It is a regional specialty oil in the south-eastern part of Europe. In this article, we describe the production and economic value of this edible oil as well as its composition on a molecular basis, including fatty acids, vitamins, phytosterols, minerals, polyphenols, and the compounds responsible for its pigments, taste and flavor. We also describe contaminants of Styrian pumpkin seed oil and the most relevant field pests of the Styrian oil pumpkin plant. Finally, we review the putative beneficial health effects of Styrian oil pumpkin seeds and of their products.
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The present studies are related to the effects of oral administration of lead acetate on rat spermatogenesis. In addition to the observations of greatly inhibited sperm motility and a reduced sperm count in the lead acetate-fed rats, as obtained in our earlier studies [Leblond et al.: Physiol Rev 52:198–236, 1972], it was considered important to study the cell kinetics in the seminiferous epithelium, a site for the generation of sperm. Lead-induced shrinkage of the seminiferous tubules was observed with a significant decrease in tubular diameter and germinal height. An overall decrease in all cell types was seen with maximum lead acetate effects on the fully mature spermatids. A ratio between the number of cells at various sequential stages was also determined. The ratios of the young spermatids to the pachytene spermatocytes and the mature spermatids to young spermatids were significantly low in the different lead-treated groups. Some depression of spermatogenesis at the young spermatid stage was observed at higher lead acetate doses. However, the arrest of spermatid maturation at the lowest dose of lead was complete. The maximum toxicity at the lowest dose of lead probably relates to the bioavailability of lead, which may be checked by the induction of lead-binding proteins as the higher lead levels. © 1996 Wiley-Liss, Inc.
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The effect of zinc deficiency on memory function, oxidative stress and blood chemistry in rats was investigated .Forty-two male (65.0±2.78g) albino Sprague Dawley rats were equally divided into three groups 4 A. B and C. Two experimental diets were formulated, consisting of 4mg ZnSO /kg as zinc deficient diet and 4 4 30mg ZnSO /kg , as Zn adequate diet. The control or commercial diet contained 15mg ZnSO /kg . Each group was respectively fed Zn deficient diet, Zn adequate diet and control diet. The rats were fed the respective diet daily and ad libitum with water for 30days. Body weights, feed and water intake of the rats were computed every other day. A shuttle box was used for the test , short term and long term memory of these rats fed the respective diets. Two criteria were considered as behavioral test, these include latency in entering dark chamber and time spent in the light chamber before entering the dark chamber at days 2, 9, 16, and 23 after the first day of training using the shuttle box. After the memory testing was conducted, rats were sacrificed by decapitation, blood was taken and brain excised. Total brain protein, plasma creatinine level, specific activities of reduced glutathione, acetylcholinesterase, superoxide dismutase, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were determined. The control diet supported significantly (p< 0.01) higher body weight gain, feed intake and water intake than zinc adequate diet. The zinc adequate rat spent the longest time in the light compartment, followed by the control rat. The specific activity of AchE of zinc adequate rats was significantly higher than rats fed zinc deficient and control diet. Brain specific activities of GSH and SOD of zinc adequate rats were significantly lower as compared with zinc deficient or control diet fed rats. There was no significant difference in the activities of plasma AST, ALT and ALP, creatinine level of rats fed zinc deficient, zinc adequate and control diets. Data of the present study indicate the effectiveness of the zinc adequate diet in memory retention and alleviation of oxidative stress in rats.
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Pumpkin seeds have been implicated in providing health benefits. However their antioxidant or anti-inflammatory activity of their extracts has never been studied. Therefore, four commercially available pumpkin seeds were treated with two different extraction methodologies in order to obtain fractions with different content. The extracts were screened for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and for their inhibitory activity against lipid peroxidation catalyzed by soybean lipoxygenase.Most extracts tested have demonstrated radical scavenging activity, which depends on their total phenolic content, with fractions rich in phenolics showing the strongest activity. On the other hand, the phenolic content of extracts does not determine their activity against lipoxygenase, as acetone and polar lipid fractions are its strongest inhibitors. The presence of molecules being able to scavenge radicals and inhibit lipoxygenase in pumpkin seeds may in part explain the health benefits attributed to them.
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