Protective effect of Chufa Tubers (Cyperus esculentus ) on induction of sperm abnormalities In mice treated with lead acetate

Abstract

The effect of the extract of Cyperus esculentus on the changes in sperm count, activity, morphology and testicular histology induces by lead acetate was studied.18 adult male rats were randomly divided into three experimental groups. Group 1 (control) were given clean drinking water and rat chow, group 2 were given intraperitoneally injection 8mg/kg body weight + water + rat chow. Group 3 were given intraperitoneally injection 8mg/kg body weight + water with extract of Cyperus esculentu+ rat cho. The results show a destruction in the testicular histology, decreased sperm count, activity and morphology of Group 2 administered with lead acetate only, which was statistically significant (P<0.05), and there was also a dose-dependent improvement in the parameters studied in the Group 3 given extract of Cyperus esculentus. This shows that the extract of Cyperus esculentus has a regenerative effect on the destroyed testicular histology induced by lead acetate.

Full text

Protective effect of Chufa Tubers (Cyperus esculentus ) on induction
of sperm abnormalities In mice treated with lead acetate
Mahera N Al-Shaikh*; Tala. A. L. Abdul Wahab**; Sahar H Abdul Kareem**;
Salim R Hamoudi ***
* Prof. Dr. Head of Medical Biology, Department of Anatomy, Medical College, University of Baghdad
**Lecturer, Department of Basic Science, College of Dentistry, University of Baghdad
**** Prof. Dr. Head of Department of Pathology, Medical College, University of Baghdad
Key words:
Cyperus esculentus. Tests. Lead acetate
How to Cite this Paper:
Mahera N Al-Shaikh, Tala. A. L. Abdul
Wahab, Sahar H Abdul Kareem, Salim R
Hamoudi “Protective effect of Chufa Tubers
(Cyperus esculentus ) on induction of sperm
abnormalities In mice treated with lead acetate” Int.
J. Drug Dev. & Res., April-June 2013, 5(2): 387-392.
Copyright © 2013 IJDDR, Mahera N Al-
Shaikh et al. This is an open access paper
distributed under the copyright agreement with
Serials Publication, which permits unrestricted use,
distribution, and reproduction in any medium,
provided the original work is properly cited.
INTRODUCTION
Lead is a heavy soft metal, occurs in nature as an
oxide or salts .Due to the industrial processes and
smokes from petrol vehicle (1) lead is considered as
one of the major environmental pollutants and
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Abstract
The effect of the extract of Cyperus esculentus on
the changes in sperm count, activity, morphology
and testicular histology induces by lead acetate
was studied.18 adult male rats were randomly
divided into three experimental groups. Group 1
(control) were given clean drinking water and rat
chow, group 2 were given intraperitoneally
injection 8mg/kg body weight + water + rat chow.
Group 3 were given intraperitoneally injection
8mg/kg body weight + water with extract of
Cyperus esculentu+ rat cho . The results show a
destruction in the testicular histology, decreased
sperm count, activity and morphology of Group 2
administered with lead acetate only, which was
statistically significant (P<0.05), and there was
also a dose-dependent improvement in the
parameters studied in the Group 3 given extract of
Cyperus esculentus. This shows that the extract of
Cyperus esculentus has a regenerative effect on the
destroyed testicular histology induced by lead
acetate.
*Corresponding author, Mailing address:
Mahera Noori Al-Shaikh
Iraq – Baghdad – College of Medicine –
Bab Al- Mouatham
Email: maheraalshaikh@yahoo.com
Article History:
------------------------
Date of Submission: 22-04-2013
Date of Acceptance: 30-05-2013
Conflict of Interest: NIL
Source of Support: NONE
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Int. J. Drug Dev. & Res., April-June 2013, 5 (2): 387-392
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387

industrial pollution (2, 3). Lead acetate is used as
water repellant for mildew protection and as a
mordant for cotton dyes. Occupational lead exposure
may occur during the manufacture of batteries,
painting, printing, pottery glazing, and lead smelting
processes .all sources of lead contribute to an
increased in permissible exposure limit for metallic
lead, lead oxide, and lead salts and soaps that has set
by WHO and other health organizations (4).
Lead accumulates virtually in every tissue of the body
and effects almost all the body systems specially
RBCs, liver, nervous system, gonads and kidneys.
Blood lead level is the most reliable indicator of lead
intoxication. Neurotoxicity and cognitive loss is a
well known side effect of lead even at low does. The
oxidative stress of lead causes damage to all
components of the cell including cell membrane,
proteins and nucleic acids (5).
Literature survey so far available indicates that lead
treatment causes anemia (6). Testicular
histochemical changes and spermatogenic inhibition
have also been observed after lead administration in
rats (7). The use of herbs in the treatment of different
diseases is fast becoming revolutionalized. In some
countries, it has been integrated into the health
scheme despite advances in orthodox medicine. It is
believed that natural products if utilized in the
correct form and dosage are less harmful than
synthetic products, which most often elicit some
anaphylactic response or reaction (8).
Cyperus esculentus
Cyperus esculentus (or chufa sedge, nut grass, yellow
nutsedge, tigernut sedge, or earth almond) is a crop
of the family (Cyperaceae) native to warm temperate
to subtropical regions of the Northern Hemisphere
.The tubers are edible, with a slightly sweet, nutty
flavour, compared to the more bitter-tasting tuber of
the related Cyperus rotundus (purple nutsedge).
They are quite hard and are generally soaked in water
before they can be eaten, thus making them much
softer and giving them a better texture (9).
In ayurvedic medicine tigernuts are used in the
treatment of flatulence, diarrhoea, dysentery, debility
and indigestion.[10] Tigernut oil can be used in the
cosmetic industry. As it is antidioxide (because of its
high content in vitamin E) it helps slow down the
ageing of the body cells. It favours the elasticity of the
skin and reduces skin wrinkles.[11].Therefore, this
study is designed to confirm the effect of lead acetate
on the histology of the testes and a possible testicular
regenerative effect of Cyperus esculentus Extract.
MATERIALS AND METHODS
The present study was conducted with 20 mature
male albino mice, weighing about 25-35 gm was
isolated in a relatively controlled environmental
temperature of about 25 °C. in the animal breeding
center / college of medicine / university f Baghdad.
Food & drinking water were available ad labium , 8
mg of Lead acetate (12) dissolved in 1 mL of distilled
water was administered in a single intraperitoneal
injection to adult mice. 18 animals divided into 3
groups: Group I control group received single
intraperitoneally normal saline only.Group II
received one injection intraperitoneally of lead
acetate (8 mg) only, while group III received lead
acetate and herbs extract of Cyprus esculentus.
Cyprus esculentus seed were purchased from a local
market in Syria. It was rinsed to remove sand and
other debris. Soak two tablespoons of cyperus for the
whole night in the water, then mashed and filtered
(13), drinking with water for period of 4 weeks.
Animals were killed on 15th and 30 days of treatment
and the testes weighed, testes were dissected out for
histopathological study.
The epididymes were excised and minced with fine
scissors in physiological saline in a Petri dish. Smears
were made on clean, grease free slides .The slides
were air-dried and coded for subsequent examination
under microscope for morphological abnormalities
according to (14). Champers of haemocytometer was
charged with 5µI of the fluid, then the sperms were
counted and the total numbers of sperms per mm
3
Full Le n g t h R e s e a r c h M an u s c r i p t
Mahera Noori Al
-
Shaikh
et al:
Protective effect of Chufa Tubers (
Cyperus esculentus
) on
induction of sperm abnormalities In mice treated with lead acetate
Int. J. Drug Dev. & Res., April-June 2013, 5 (2): 387-392
Covered in Scopus & Embase, Elsevier
388

were calculated. The data was expressed as Mean ±
SD. The differences between groups were compared
for statistical significance by t-test with the level of
significance set at P≤ 0.05.
Results
Light microscopy was used for evaluation of
testicular histology and the seminiferous tubules of
the control and different experimental groups shows:
control group demonstrated normal somniferous
tubules with presence of spermatogony maturation,
spermatogenesis was evident in most tubules, and
leydic cells are normal (fig. A).
A B C
In contrast, in the lead acetate treated group, the
tests showed an abnormal structure of the
seminiferous tubules with vacuolar degenerative
changes appearing in the cytoplasm of the
spermatogenic epithelium and absence of late stage
germ cells showed damage and sloughing of
somniferous tubules (Fig. B).Testicular section
from animal received lead acetate and treated with
herbal extract showed a slight change from normal
histological features, somniferous tubules with
maturation of spermatogony and sperms inside the
lumen (Fig. C ).
Testicular weight and sperm count was significantly
decreased in experimental groups compared with
normal control group (table I).
Table 1: Changes in body weight and testicular
sperm count
Groups
Testicular
2 week
Weight
4 weeks
sperm
2 week
Count
4weeks
Control
0.65 ±
0.04 0.62±0.02
200 ±
0.2
195 ±
0.02*
Lead acetate 0.23±0.2*
0.3±0.4*
90 ±
0.4*
100 ±
0.2*
Lead acetate
+herbal extract
0.52±0.2
0.45±0.2*
150 ±
0.2*
140 ±
0.2
P – Values * <0.05 compared with the
corresponding vehicle treated controls.
The results of sperm head abnormality counts
showed that there was a high level of abnormality
in the sperm head of mice treated with lead acetate
compared with control (A). These abnormalities
observed are : hook at wrong angle (B ) ,spiral head
( C) , pin-head ( D) , two tails ( E ) , head only (F ) ,
sperm with no hook (F ) , banana-shaped head ( D)
, amorphous head ( D, H ) , mean bent hook (G ) .
A B C
Full Le n g t h R e s e a r c h M an u s c r i p t
Mahera Noori Al
-
Shaikh
et al:
Protective effect of Chufa Tubers (
Cyperus esculentus
) on
induction of sperm abnormalities In mice treated with lead acetate
Int. J. Drug Dev. & Res., April-June 2013, 5 (2): 387-392
Covered in Scopus & Embase, Elsevier
389

D E F
G H
I
DISCUSSION
Lead (Pb+2) is a heavy metal that can be toxic when
introduced into the human and animal bodies by
ingestion or inhalation in sufficient quantities. It
causes various destructive effects (15). In human,
increased levels of lead causes many serious diseases
and dysfunction of organs (16).Lead is well known to
produce oxidative damage by enhancing lipid
peroxidation (17) Lipid peroxidation inactivates cell
constituents by oxidation or causes oxidative stress
by undergoing radical chain reaction, ultimately
leading to loss of membrane integrity (18). In
analogy, lead was shown to implicate testicular lipid
peroxidation as evidenced by previous reports (19)
this could be due to the formation of free radicals
(18) through an exhaustion of antioxidants (20) and
subsequently to oxidative stress (21). ). On the other
hand, since lead does not undergo oxidation-
reduction cycle, the effect of lead on lipid
peroxidation is not a direct effect, but these changes
could rather be due to an indirect effect of lead on the
free-radical scavenging enzymes( 22) and/or
glutathione peroxidase (20). However, the higher
membrane lipid content of testes is presumed to
make them more vulnerable to oxidative stress (23).
Full Le n g t h R e s e a r c h M an u s c r i p t
Mahera Noori Al
-
Shaikh
et al:
Protective effe
ct of Chufa Tubers (
Cyperus esculentus
) on
induction of sperm abnormalities In mice treated with lead acetate
Int. J. Drug Dev. & Res., April-June 2013, 5 (2): 387-392
Covered in Scopus & Embase, Elsevier
390

The elevation of frequency of sperm head
abnormalities in mice treated with lead acetate may
have caused damage to the pre-meiotic stages of
spermatogenesis since during spermatogenesis, DNA
synthesis occurs before pre-meiotic phase and no
further DNA synthesis occurs throughout
spermatogenesis in the cell cycle (24, 25).Several
reasons have been put forward to explain the
increase in the frequency of occurrence of sperm
head abnormalities in organisms exposed to some
chemicals. In general, damage to the sperm cell is
said to occur either by physiological, cytotoxic or
genetic mechanism. Odeigah (24) reported that
exposure to the chemicals could produce pituitary
hypothalamic or sex hormonal effects which in turn
could affect spermatogenesis or exposure could cause
abnormalities in seminal fluid resulting in functional
or structural impairment of sperm . It may also arise
as a consequence of naturally occurring level of
mistakes in the spermatozoon differentiating process
during spermatogenesis (26). In this study, Treated
with Cyperus esculentus relatively improves the
induced histopathological changes in the testis of
rats caused by effect of lead acetate, these
improvement may be either : directly on
spermatogenesis by reducing lipid peroxidation and
prevent or decrease the formation of free radicals ,
by acting as antioxidant . Or indirectly through
pituitary hypothalamic or sex hormonal effects.
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Protective effect of Chufa Tubers (
Cyperus esculentus
) on
induction of sperm abnormalities In mice treated with lead acetate
Int. J. Drug Dev. & Res., April-June 2013, 5 (2): 387-392
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Mahera Noori Al
-
Shaikh
et al:
Protective effect of Chufa Tubers (
Cyperus esculentus
) on
induction of sperm abnormalities In mice treated with lead acetate
Int. J. Drug Dev. & Res., April-June 2013, 5 (2): 387-392
Covered in Scopus & Embase, Elsevier
392