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* Corresponding author, Address: Iran, Jahrom,Jahrom Branch, Islamic Azad University, Young Researchers
and Elit Club,
Tel: 09171095411 Email: [email protected] 45
Protective effect of green tea extract on ovary tissue function in rats treated
by Malathionin secticide
Hoseini Shahrkhafri Marziyah Sadat٭1, Hemayatkhahjahromi Vahid
2, Samani Jahromi
Elaheh1
Received:4/19/2015 Revised:12/14/2015 Accepted:12/20/2015
1. Young Researchers and Elit Club, Jahrom Branch, Islamic Azad University, Jahrom, Iran
2. Department of Biology, Jahrom Branch, Islamic Azad Unversity, Jahrom, Iran
Pars Journal of Medical Sciences, Vol. 13, No.3, Fall 2015
Abstract
Introduction:
This study was conducted to investigate the protective effect of green tea extract on ovary tissue
function in rats treated by Malathion insecticide.
Materials and Methods:
A total of 72 mature Wistar rats aged 2 to 3 months (approximate weight of 200±15 g) were
experimented. Rats were divided into 9 groups of eight. Control group did not receive any medication.
Sham group was given 0.2 cc physiological serum and experimental groups 1, 2 and 3 received 100,
200 and 400 mg/KG bw green tea extract respectively, experimental 4 was given 40 mg/KG bw
Malathion and experimental groups 5, 6 and 7 received 100, 200 and 400 mg/KG bw green tea extract
respectively and 40 mg/KG bw Malathion. After 15 days, the serum levels of sex hormones were
measured, and ovaries were removed for counting the ovarian follicles.
Results:
Serum concentration of estrogen, progesterone, FSH and LH were significantly decreased in
experimental group 4 compared to control and other groups. The primary and Graafian follicles
decreased in experimental group 4 compared to control group. Primary and secondary follicles and
corpus luteum significantly decreased in experimental group 4 compared to control group. All
examined parameters except Graafian follicles and corpus luteum increased in the experimental
groups 5, 6 and 7 compared to experimental group 4. Follicular atresia increased in experimental
group 4 compared to other groups and significantly decreased in experimental groups 5, 6 and 7
compared to experimental group 4.
Conclusions:
Malathion had an adverse effect on secretion of sex hormones in female rats, as well as the process of
oogenesis, and green tea extract decreased those negative effects.
Keywords:Malathion, Green Tea, Oogenesis, Sex Hormones, Rat
Introduction
Green tea is currently considered a useful
source for human health with biological
and pharmacological activities. The health
benefits of tea extract and its polyphenolic
catechins have attracted most scientific
research in the prevention and treatment of
many diseases. Tea leaves, as the only
food product having Epigallocatechin -3-
Th
Par J Med Sci 2015;13(3):45-57
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
46
gallate (EGCG) have active compounds
with eight free OH-groups and high
antioxidant activity (1, 2). Tea is called
khatai tea in traditional medicine and Shay
in Arabic, and its shrub or bush is called
Theier in French and Teaplant in English.
This plant belongs to Temsnstroemiaceae,
Camelliaceae, and Theaceae families, and
Camellia sinensis and Camellia Theifer are
its scientific names. Tea is a big shrub with
a maximum height of one and a half
meters and glossy leaves and beautiful
white flowers. This plant contains
polyphenolic compounds, including
Epigallocatechin -3- gallate (EGCG),
Epigallocatechin -EGC, Epicatechin -3-
gallate-ECG, and Epicatechin –EC (3, 4, 5,
6). The most potent of these antioxidants,
Epigallocatechin -3- gallate (EGCG) has
been reported to be 100 times as potent as
vitamin C and 25 times as potent as
vitamin E (7). The protective effect of
green tea on body tissues may be due to its
antioxidant property (8). The most
important ingredient in green tea is
catechin (ECGC, the most abundant
catechin). This substance is an extremely
potent and effective antioxidant and many
beneficial effects of green tea are
attributed to it (9, 10, 11). Studies have
shown that a cup of green tea (2.5 g green
tea in 200 ml of water) may contain 90 mg
EGCG (12). Green tea has been introduced
as an anti-inflammatory, anti-cancer, anti-
cholesterol, anti-diabetic, anti-mutation,
anti-microbial, anti-stroke, and anti-
oxidative agent (13). In addition, bioactive
components of green tea (catechins and
caffeine) stimulate the sympathetic system
and increase heat production and fat
oxidation in the body and accordingly,
exert their anti-obesity effects (14, 15).
Catechin, as a natural antioxidant, affects
the reproductive system and stimulates
testosterone production by the Leydig cells
and decreases the content of total
cholesterol, triglycerides and
phospholipids in testicular tissue. It also
prevents the mutagenicity of mutagenic
chemicals on chromosomes (8). The
ability of sperm viability, mobility, and
fertilization highly depends on its
antioxidant capacity expressed in seminal
plasma. Being rich in polyphenols, as
potent antioxidants, green tea inhibits
reactive oxygen and nitrogen species, and
thus improves sperm quality (9).
Malathion (S-(1,2-dicarboethoxyethyl) O,
O-dimethyl phosphorodithioate) is one of
the most widely used insecticides in Iran.
Malathion is a yellowish dark brown
concentrated and oily liquid that is used as
a broad-spectrum pesticide in agriculture
(16). This substance is a derivative of D-
tonic phosphoric acid and one of the oldest
P-group insecticides introduced in 1950
(17). Organophosphates (OPs) are a class
of pesticides widely used in agriculture,
veterinary medicine, industry, residential
areas, and as nerve agents in wars. These
compounds cause the most accidental
poisonings with acute or chronic
complications (18, 19).
Studies have shown that toxic effects of
some OPs are not limited to cholinesterase
inhibition, but as a result of cholinergic
crisis (increased acetylcholine), changes
such as cell membrane damage, free
radical production, and the antioxidant
system malfunction are also observed (20,
21). By conducting a study on the
genotoxic effects of Malathion, Giri et al.
(2002) found that this toxin affects sperm
count and causes cell mutation (22).
Weisburger (2002) reported that tea
polyphenols have potent antioxidant
activity and can reduce LDL oxidation and
oxidized DNA metabolism, and
subsequently decrease the risk of heart
diseases and cancer (13). Crespy and
Williamson (2004) reported that green tea
extract has antioxidant properties and
eliminates free radicals (23). Since green
tea is high in catechins, it is considered a
potent antioxidant. Azarnia et al. (2013)
conducted a study on the impact of
hydroalcoholic extract of green tea on rats
with polycystic ovary syndrome (PCOS)
and reported reduced serum concentrations
of LH, CRP, IL-6, insulin, and glucose in
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
47
groups treated with green tea extract.
Histomorphometric studies also indicate
an increase in the number of follicles and
changes in the thickness of the follicular
theca cells for improving PCOS. Green tea
can be a potential medication for the
treatment of PCOS and type 2diabetes
(24).
The results of the study of Sanaei et al.
(2014) also showed that administration of
green tea extract for 42 days significantly
increased the number of primary, growing,
and antral follicles (25). Shariatzadeh and
Mohammadi (2014) reported that the use
of green tea can significantly compensate
damage caused by the use of sodium
arsenite, sperm count, motility, viability,
and normal morphology, as well as the
diameter of the lumen of the seminiferous
tubules, germinal epithelial thickness, and
the levels of MDA (P<0.001) (26).
The present study aimed to investigate the
protective effect of green tea extract on
ovary tissue function in rats treated by
Malathion insecticide.
Materials and Methods
Animals used: A total of 72 mature
female Wistar rats aged 2 to 3 months
(approximate weight of 200±15 g) were
purchased from Animal House of School
of Veterinary Medicine, Shiraz, Iran. Rats
were transferred to the Animal House of
Jahrom University of Medical Sciences
and were housed in special sterilized
containers at a temperature of 22-20 °C
and moisture content of 60-40 percent for
two weeks to be adapted to the
environment. During the experiment, they
were given foods provided from Fars
Livestock and Poultry Company and tap
water in special drinkers.
Medications and materials used: Green
tea plant (medicinal herb shops in the
city), Malathion (Moshkfamfars factory),
the FSH hormone kit (Iran Pishtaz Teb,
serial number Catnom BYEK1370), the
LH hormone kit (Iran Pishtaz Teb, serial
number Catnom BYEK1370), the
progesterone hormone kit (REF DKO 006,
Germany), the estrogen hormone kit (REF
DKO 003, Germany).
Green tea extraction method: To prepare
the extract, green tea was first milled and
soaked in 80% ethanol for 24 hours. The
solution was then filtered and the extract
was obtained through vacuum
concentration method. The extract will be
soluble in distilled water (27, 2).
Methods: This experimental study was
conducted in Animal House of Islamic
Azad University, Jahrom Branch, May
2013. All ethical considerations were
observed in relation to animal care and
working with laboratory animals during
the study and the topic was approved by
the Ethic Committee of Islamic Azad
University, Jahrom Branch (D/P/2990
dated February 28, 2013). According to
previous articles, rats were divided into 9
groups of eight as follows:
Control group did not receive any
medication. Sham group was given only
physiological serum. Experimental groups
1, 2, and 3 received 100, 200, and 400
mg/kg bw green tea extract respectively,
experimental 4 was given 40 mg/kg bw
Malathion, and experimental groups 5, 6,
and 7 received 100, 200, and 400 mg/kg
bw green tea extract respectively and 40
mg/kg bw Malathion. Rats were injected
intraperitoneally with green tea extract and
Malathion for 14 days.
Determination of serum levels of
hormones: On the 15th
day, rats were
anesthetized with diethyl ether and blood
samples were taken from their hearts.
Blood samples were placed in a steam bath
at 37 °C to be clotted. After centrifugation
at 3000 rpm for 10 minutes, the serum was
separated. LH, FSH, progesterone, and
estrogen levels were measured using the
ELISA method. After the sampling, the
abdominal region of the rats was incised
with a scalpel. Left and right ovaries were
removed with a scalpel and forceps from
the surrounding fat tissues and the
fallopian tube and after washing with
physiological saline, they were placed in
separate containers of 3% formalin
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
48
solution. Tissue sections were then
prepared from the samples. Prepared slides
of left and right ovaries of rats were
studied separately by a light microscope.
In each slide, the number of primary,
secondary, Graafian, and atretic follicles,
and corpus luteum were counted with 40x
and 100x magnification.
Statistical analysis method: One-way
analysis of variance (ANOVA) was used
for statistical analysis. Since the data were
normally distributed according to the
Kolmogorov-Smirnov test results,
parametric tests were used. Duncan’s test
was applied to find the difference between
the means when there was a significant
difference between different groups.
Statistical analysis was performed using
SPSS-16 and the significance level was
considered P<0.05. The results are
presented as Mean±SEM in the results
section. Figures were drawn by Excel
software.
Results
Changes in plasma concentration of LH:
The results of changes in plasma
concentrations of LH showed that LH
levels reduced significantly in
experimental group 4 (Malathion group)
compared to control group and other
experimental groups at P<0.05. A
significant increase was observed in
experimental groups 5, 6, and 7 (groups
receiving minimum, average, and
maximum Malathion and tea) compared to
Malathion group at P<0.05 (Figure 1).
Figure 1: Comparison of LH concentrations between the groups studied
The values are shown as Mean±S.E.
According to Duncan’s test, if columns have at least one common letter, it indicates there is no
significant difference between columns.
Changes in plasma concentration of
FSH:
The results of changes in plasma
concentrations of FSH among the groups
studied showed that FSH levels reduced
significantly in experimental group 4
(Malathion group) compared to control
group and other experimental groups at
P<0.05. A significant increase was
observed in experimental groups 5, 6, and
7 (groups receiving minimum, average,
and maximum Malathion and tea)
compared to Malathion group at P<0.05
(Figure 2).
b
b
b
b
b
a
b
b b
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Ho
rmo
ne
(LH
(IU
/L)
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
49
Figure 2: Comparison of FSH concentrations between the groups studied
Changes in plasma concentration of
estrogen:
The results of changes in plasma
concentrations of estrogen among the
groups studied showed that estrogen levels
significantly increased in experimental
group 2 (group receiving average dose of
tea) and significantly decreased in
experimental group 4 (Malathion group)
compared to control group at P<0.05. A
significant increase was also observed in
experimental groups 5, 6, and 7 (group
receiving minimum, average, and
maximum Malathion and tea) compared to
Malation group at P<0.05 (Figure 3).
Figure 3: Comparison of estrogen hormone between the groups studied
Changes in plasma concentrations of
progesterone:
The results of changes in plasma
concentrations of progesterone among the
groups studied showed that progesterone
levels significantly decreased in
experimental groups 4 (Malathion group)
and 5 (the group receiving Malathion and
the minimum tea) and significantly
increased in experimental group 2 (group
receiving average dose of tea) compared to
control group (P<0.05). A significant
b b
b
b b
a
b
b b
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lasm
a co
nce
ntr
atio
n o
f FS
H(I
U/L
)FSH
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
bcd bcd
de
e
cd
a
bc bc
b
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ho
rmo
ne
estr
oge
n.
))n
g/m
l
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
50
increase was also observed in progesterone
levels in experimental groups 5, 6, and 7
(group receiving minimum, average, and
maximum Malathion and tea) compared to
Malation group at P<0.05. A significant
increase was also observed in plasma
concentrations of progesterone in
experimental groups 6 and 7 compared to
experimental group 5 (Figure 4).
Figure 4: Comparison of progesterone between the groups studied
The number of primordial follicles:
The results of the number of primordial
follicles among the groups studied showed
that the number of primordial follicles
increased in experimental groups 2 and 3
(the group receiving average and
maximum doses of tea) and significantly
decreased in experimental group 4
(Malathion group) compared to control
group (P<0.05). A significant increase was
also observed in experimental 5, 6, and 7
(the group receiving minimum, average,
and maximum Malathion and tea)
compared to Malathion group (P<0.05)
(Figure 5).
Figure 5: Comparison of the number of primordial follicles between the groups studied
d
cd
d
e
d
a
b
c c
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control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
c bc
c
d d
a
bc bc bc
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۱۲.۰۰۰۰
Pri
mo
rdia
l fo
llicl
e(%
)
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
51
Changes in the number of primary
follicles:
The results of the number of primary
follicles among the groups studied showed
that the number of primary follicles
significantly decreased in experimental
group 4 (Malathion group) compared to
control group and other groups (P<0.05).
A significant increase was also observed in
experimental 5, 6, and 7 (the group
receiving minimum, average, and
maximum Malathion and tea) compared to
Malathion group (P<0.05) (Figure 6).
Figure 6: Comparison of the number of primary follicles between the groups studied
Changes in the number of secondary
follicles:
The results of the number of secondary
follicles among the groups studied showed
that the number of secondary follicles
decreased in experimental group 4
(Malathion group) and significantly
increased in experimental group 2 (average
dose of tea) compared to control group
(P<0.05). A significant increase was also
observed in experimental 5, 6, and 7 (the
group receiving minimum, average, and
maximum Malathion and tea) compared to
Malathion group (P<0.05) (Figure 7).
Figure 7: Comparison of the number of secondary follicles between the groups studied
c c
bc
c c
a
bc bc
ab
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۶.۰۰۰۰
nu
mb
er o
f p
rim
ary
folli
cles
(%)
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
b bc bc
c
bc
a
ab ab
ab
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۷.۰۰۰۰
۸.۰۰۰۰
۹.۰۰۰۰
۱۰.۰۰۰۰
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
52
Changes in the number of Graafian
follicles:
The results of the number of Graafian
follicles among the groups studied showed
that the number of Graafian follicles
increased in experimental groups 2 and 3
(the group receiving average and
maximum doses of tea) and significantly
decreased in experimental group 4
(Malathion group) compared to control
group (P<0.05). A significant increase was
also observed in experimental 5, 6, and 7
(the group receiving minimum, average,
and maximum Malathion and tea)
compared to Malathion group (P<0.05)
(Figure 8).
Figure 8: Comparison of the number of Graafian follicles between the groups studied
Changes in the number of corpus
luteum:
The results of the number of corpus luteum
among the groups studied showed that the
number of corpus luteum increased in
experimental group 2 (average dose of tea)
and significantly decreased in
experimental groups 3 (maximum dose of
tea) and 4 (Malathion group) compared to
control group and other groups (P<0.05).
A significant increase was also observed in
experimental 5, 6, and 7 (the group
receiving minimum, average, and
maximum Malathion and tea) compared to
Malathion group (P<0.05) (Figure 9).
Figure 9: Comparison of the number of corpus luteum between the groups studied
c c
cd
d d
a
ab
bc
ab
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۰.۵۰۰۰
۱.۰۰۰۰
۱.۵۰۰۰
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۳.۰۰۰۰
۳.۵۰۰۰
۴.۰۰۰۰
۴.۵۰۰۰
۵.۰۰۰۰
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
bc
bc
cd
e
d
a ab
bcd abcd
۰.۰۰۰۰
۱.۰۰۰۰
۲.۰۰۰۰
۳.۰۰۰۰
۴.۰۰۰۰
۵.۰۰۰۰
۶.۰۰۰۰
nu
mb
er o
f co
rpu
s lu
teu
m(%
)
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
53
Changes in the number of atretic
follicles:
The results of the number of atretic
follicles among the groups studied showed
that the number of atretic follicles
significantly increased in experimental
group 4 (Malathion group) compared to
control group and other groups (P<0.05).
A significant decrease was also observed
in experimental 5, 6, and 7 (the group
receiving minimum, average, and
maximum Malathion and tea) compared to
Malathion group (P<0.05) (Figure 10).
Figure 10: Comparison of the number of atretic follicles between the groups studied
Discussion
The results showed that progesterone,
estrogen, FSH, and LH levels significantly
reduced in experimental group 4 compared
to control group and other groups, and
significantly increased in experimental
groups 5, 6, and 7 (the group receiving
minimum, average, and maximum
Malathion and tea) compared to Malathion
group (P<0.05). Increased serum levels of
sex hormones have been reported useful
for the evaluation of human and animal
fertility (28). In general, a significant
reduction in concentration of sex
hormones in fertility activities leads to
fertility disorders in people who are
exposed to chemicals (29).
Exposure to chemicals led to ovarian
tissue destruction and significant changes
in progesterone and estrogen in rats which
it was also confirmed in previous studies.
Yarbe et al. (2009) reported that reduced
serum levels of progesterone and estrogen
occur due to ovarian damage which is
consistent with the results of this study. At
the time of fertility and ovulation, estrogen
and progesterone are normally affected by
the pituitary gland producing LH and FSH
(30). Thus, it can be said that ovarian
tissue damage and impaired secretion of
estrogen and progesterone which can occur
increasingly or decreasingly justify a
significant reduction in estrogen and
progesterone in experimental group 4
compared to control group. LH and FSH
changes may also be due to reduced GnRH
which decreased the serum levels of LH
and FSH. The hypothalamic-pituitary-
gonadal axis regulates reproductive
activities, a multifactorial process, by
genetic and hormonal control. In this
study, the results of histological evaluation
showed that treatment with Malathion
leads to ovarian cell damages and induces
extensive histopathological changes in
ovarian follicles. Malathion can cause
toxic effects on the reproductive system
through oxidative stress (31), which can be
reduced with concurrent administration of
antioxidants. This result is consistent with
our results. By conducting a study on the
a a a
a a
c
b b
b
۰.۰۰۰۰
۱.۰۰۰۰
۲.۰۰۰۰
۳.۰۰۰۰
۴.۰۰۰۰
۵.۰۰۰۰
۶.۰۰۰۰
۷.۰۰۰۰
nu
mb
er o
f at
reti
c fo
llicl
es(%
)
control shahed Tea mini. Tea average Tea max. malathion malt & T.min malt&T.aver malt&T.max
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
54
protective effect of green tea extract on
ovary function in mice treated by
Paclitaxel drug, Mohseni Kouchesfehani
and Asoudeh (2014) reported that green
tea extract improves ovarian parameters
treated by Paclitaxel. Green tea extract
with its antioxidant properties has
protective effects on ovarian tissue
parameters in mice and LH and FSH levels
after treatment with Paclitaxel (32). This
protective effect of green tea on the
parameters of ovarian tissue is consistent
with our results. According to a study
conducted by Ghafuriyan et al. on the
effect of green tea extract on the histology
of the ovary in polycystic ovary syndrome,
it was found that green tea affects oocyte
maturation and follicular development in
polycystic women. Green tea extract
reduces the theca layer thickness in rats
and may increase lipolysis and reduce the
hypertrophy of this layer. Accordingly, the
production of androgens and steroids also
decreases by the mentioned layer (33). It
seems that the difference in the type of
animal, developmental stage, treatment
duration, and the type of field is the main
reason for these differences. The results of
the study of Allahdadian et al. showed that
green tea consumption has a significant
effect on reducing free testosterone in
patients with polycystic ovarian disease
(34). According to a laboratory study
conducted by Figueiroa et al. on the effect
of green tea compounds on testosterone
production in rabbit Leydig cells, it was
found that green tea compounds inhibit
basic and stimulated production of
testosterone (35). According to a study
conducted by Wu et al. on the effect of
two months of green tea consumption on
testosterone level in healthy
postmenopausal women, it was reported
that green tea supplements caused no
constant changes in the testosterone levels
(36). According to a study conducted by
Shayeghi et al. on the effect of Malathion
insecticides on the inhibition of
cholinesterase enzyme among the
agricultural sprayers, it was found that
Malathion, as a phosphate insecticide, has
gastrointestinal toxicity and fumigant
property. They also found that decreased
cholinesterase activity will be less if health
regulations are observed when using
Malathion (37).
Fortunato et al. (2006) reported that
Malathion induces the production of free
radicals and oxidative stress in the brain
and increases the activity of antioxidant
enzymes (31). Oxidative stress is caused
by free radicals and mitochondria are
identified the main region of the
production of free radicals (38). By
evaluating the effect of Malathion-induced
stress on fat metabolism, scientists found
that the lethal and sub-lethal doses of this
toxin increase fatty acids, glycerol and
lipase activity in tissues studied and can
have a negative impact on reproductive
activity. Malathion also has a negative
impact on reproductive activity and
decreases the amphibian population which
is consistent with the present study (39).
Studies conducted in recent years
emphasize the role of reactive oxygen
species as a contributing factor in some
tissue damage. An imbalance in the
production of reactive oxygen species and
antioxidant enzyme activities leads to
tissue damage and it can occur through
either increased production of reactive
oxygen species or decreased antioxidant
enzyme activities, or both, suggesting the
existence of oxidative stress conditions
(40). Reactive oxygen species can severely
endanger cell life by weakening the
structure and function of plasma
membrane and intracellular membranes
(41). By examining ovarian tissues and
counting the follicles using a light
microscope, a decrease was observed in
the number of primordial and Graafian
follicles in experimental group 4
(Malathion group) and a significant
increase was found in experimental groups
2 and 3 (the average and maximum doses
of green tea) compared to control group
(P<0.05). The number of Graafian follicles
also increased in experimental group 6
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
55
compared to Malathion group. The number
of primordial follicles increased in
experimental groups 5, 6, and 7 (the group
receiving minimum, average, and
maximum Malathion and tea) compared to
Malathion group (P<0.05). The number of
atretic follicles significantly increased in
experimental group 4 (Malathion group)
compared to other groups (P<0.05). Also,
the number of atretic follicles in
experimental groups 5, 6, and 7 (the group
receiving minimum, average, and
maximum Malathion and tea) significantly
reduced compared to Malathion group
(P<0.05).
The number of primary and secondary
follicles and corpus luteum significantly
reduced in experimental group 4 compared
to control group (P<0.05). The number of
primary and secondary follicles
significantly increased in experimental
groups 5, 6, and 7 (the group receiving
minimum, average, and maximum
Malathion and tea) compared to Malathion
group (P<0.05).
Organophosphate insecticides have
alkylating properties and thus can affect
the cell nuclear DNA (42). These
chemicals have also electrophilic property.
Therefore, they can affect cellular proteins
and lead oocytes and follicular cells to lose
their efficiency. In this study, the
destructive impact of Malathion was also
confirmed (43). Malathion increases
malondialdehyde in ovarian tissue that
may be caused by released radicals or lipid
and body metabolism. With increasing
doses, Malathion reduces the number of
healthy follicles, increases the number of
atretic follicles, and causes changes in the
corpus luteum which is consistent with the
results of the present study (44). Salvadori
et al. (1988) found that Malathion induces
chromosomal aberrations in somatic cells
(bone marrow) and primordial germ cells
(primary spermatocytes) (45). Studies have
also indicated that oxidative stress plays an
important role in pathogenesis of various
diseases, including cancer, diabetes,
cardiovascular diseases, Parkinson's
disease, schizophrenia, atherosclerosis,
pulmonary diseases, and cataracts (46).
Oxidative stress is caused by free radicals
and mitochondria are identified in the
main region of the production of free
radicals (38). When cell properties are
changed, oocytes and follicular cells lose
their normal efficiency. According to what
was mentioned and the fact that the
production of primordial follicles in
mammals in embryonic period is a local
phenomenon caused by ovarian factors and
germ cells, and considering the fact that
the continued growth of the follicles is
influenced by the hypothalamic and
pituitary hormones, a significant decrease
in primordial, primary, and secondary
follicles can thus be justified. In this study,
significant changes in the pituitary
hormones were also observed. As
mentioned, cellular damage caused by
pesticides can be caused by various
reasons, but the impact of Malathion on
the structure of DNA and cellular proteins
can change cell function. Considering the
possible effect of Malathion on changes in
natural concentrations of gonadotropins at
the time of injection, it can be concluded
that with abnormal development of
follicular cells, reduced thickness of
granulosa layer and follicular sheath,
estrogen production decreases which is
consistent with our results. Development
of follicular sheath is directly affected by
factors secreted from the granulosa layer
and reduced granulosa layer can thus affect
the growth of follicular sheath (38).
Studies have shown that green tea has
antioxidant properties and eliminates free
radicals (23) through the inhibition of
cytochrome P450 expression (47, 48). It
has also been reported that green tea
extract can reduce oxidative damage
caused by cyclosporine A (27). Studies
have shown that green tea catechins
prevent lipid peroxidation caused by
chemicals in the liver and kidneys of
animals (49).
Protective effect of green tea extract Hoseini Shahrkhafri M.S et al
Pars Journal of Medical Sciences, Vol. 13, No. 3, Fall 2015
56
Conclusion
According to the results of histological
studies and ovarian hormones, it can be
concluded that Malathion insecticide
destroys ovarian tissue and reduces the
activity of the pituitary-gonadal hormones.
These effects can be generalized to
humans to a less extent. However, the
manner of use and extent of exposure to
this insecticide is very important.
Therefore, care should be taken in
generalizing these results to humans. Due
to its antioxidant property, green tea
extract can reduce the negative effects of
Malathion insecticide, so the use of green
tea extract is recommended to reduce the
harmful effects of Malathion.
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