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Life Science Archives (LSA)

ISSN: 2454-1354
Volume 1; Issue - 2; Year 2015; Page: 127 - 137

Research Article

HEPATOPROTECTIVE EFFECT OF Emblica Officinalis AGAINST

ENDOSULFAN AND LINDANE INDUCED CHANGES IN BIOCHEMICAL
AND ANTIOXIDANT ACTIVITY OF FRESHWATER FISH, Channa striatus
J. Prakash Sahaya Leon, M. Mariappan and K. Balakrishnan
Department of Zoology DDE, Annamalai University, Chidambaram 608 002, Tamil Nadu, India.
Abstract
Organochlorine pesticides have destroying properties on liver. Endosulfan, lindane adversely affect
the hepatic cells and induce the oxidative stress. Emblica officinalis plays a vital role to challenge many
diseases in animals and human beings. The present study investigated the induction of oxidative stress and
biochemical changes in the liver of fish Channa striatus exposed to organochlorine pesticide endosulfan and
lindane, and tried to establish the cytoprotective properties of Emblica officinalis with respect to
hepatoprotection in them. Fishes were divided into 3 groups (A C). Group A is control, Group B exposed
to 0.455 g L-1 (1/15th of 96 hr LC50) concentration of endosulfan and Group C exposed to 1.51 g L-1 (1/15th
of 96 hr LC50) concentration of lindane for 30 days. After exposure of 30 days, the fishes in the group A to C
treated with 100 mg/kg bw/day of E. officinalis for 30 days. During the experiment for each 10 days,
biochemical and antioxidant activity in liver were determined by using standard methods. The changes
recorded from exposure groups (B and C) are indicates the unhealthy conditions of fishes in endosulfan and
lindane. The exposed fishes were treated with E. officinalis, the total protein, lipid content and SOD, CAT,
GPx, GSH activities were recovered in all the three groups (A-C). This highlights the debilitating effect of
endosulfan and lindane and hepatoprotective property of E. officinalis.
Key words: Organochlorine pesticide,
Endosulfan, Lindane, Emblica officinalis and
Antioxidant activity.

Article History
Received : 26.03.2015
Revised : 08.04.2015
Accepted : 12.04.2015
1. Introduction

Pesticides have been widely used all over

the world to control insects, pests and disease
vectors and they are one of the most potentially
harmful
chemicals
introduced
into
the
environment, though they have contributed
considerably to human welfare, their adverse
effects on non-target organisms are significant.
Pesticides ultimately finds their way into aquatic
habitats such as rivers, lakes and ponds, and have
been found to be toxic to organisms, which found
* Corresponding author: J. Prakash Sahaya Leon,
Department of Zoology DDE, Annamalai University,
Annamalai Nagar, Tamil Nadu, India.

to be contributing substantially to the food chain

and in the water bodies, where fish encounter with
them
and
develop
various
metabolic
abnormalities. They accumulate in fish and affect
human health too via ecological cycling and
biological magnification.
One of the most common environmental
pollutants entering the aquatic system both in
industrial effluents and runoff from agricultural
chemicals is organochlorine pesticide. It is highly
persistent
and
non-biodegradable.
The
organochlorine insecticide endosulfan is a highly
toxic; it acts primarily on the nervous system of

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J. Prakash Sahaya Leon/Life Science Archives (LSA), Volume 1, Issue 2, Page 127 - 137, 2015

wildlife, birds, amphibia, fish and aquatic insects,

plankton, and soil microorganisms. In rainbow
trout (Oncorhynchus mykiss) endosulfan has
caused a wide range of effects including:
hyperactivity, convulsions, paralysis, erratic
behaviour, and eventually death through water
borne and feed borne exposures (Broomhall, 2002;
Naqvi and Vaishnavi, 1993). Lindane is an
organochlorine pesticide that has been widely used
in public health and agriculture production in
developed and developing countries including
India. It is used to protect crop from insects, for
pest control in forests, and in homes to control
ants and other household pests. Lindane enters
into aquatic ecosystems as a consequence of rains,
soil leaching and sewage discharges, etc.
Therefore, fish and other aquatic organisms may
show signs of its effects, although they are not the
target of this pesticide (Fanta et al., 2003).
Lindane is a potent stimulant of the central
nervous system that produces biochemical and
histological alterations in animals (Moreno et al.,
1995). Several physiological functions are
disturbed by this organochlorine insecticide during
short-term intoxication: oxygen uptake, blood
lactic acid and lipid metabolism (Bakthavathsalam
and Reddy, 1981; Gluth and Hanke, 1985).
There are many reports of correlation
between high consumption of fruit and vegetables
or of specific dietary antioxidants (vitamin C,
carotenoids, vitamin E) and a relatively low
incidence of several diseases. Dietary antioxidants
in general, act by removing reactive oxygen
species before they have chance to cause damage
to biological molecules (Khandelwal et al., 2002).
Amla (Emblica officinalis, Family Euphorbiaceae)
is a small or medium sized, deciduous tree with
smooth greenish grey, exfoliating bark. The fruits
are depressed globose, fleshy and obscurely 6
lobed, containing six trigonous seed. It has been
widely used in Indian traditional medicine for the
treatment of various diseases. Most of the
experiments conclude fruits of
E. officinalis
have antifungal, antibacterial, antidiabetic,
antipyretic,
antioxidative,
anticlastogenic,
hepatoprotective etc., (Dhir et al., 1991 & 1993;

128

Gulati et al., 1995; Ihandolla et al., 1997;

Khandelwal et al., 2002). Fruits contain gallic
acid (1.32%), tannin, sugar (36.10%), gum
(13.75%), albumin (13.08%), crude cellulose
(17.08%) and minerals (4.12%), Cr, Zn, Fe and Cu
apart from that highest amount of vitamin C
present in natural form.
Looking down its extensive therapeutic
use, limited knowledge is available regarding its
role in fish against pesticide toxicity. Few studies
have been carried out on its radical scavenging
properties and immune-modulation (Zhao et al.,
2008; Sai Ram et al., 2008). Hence, the present
study is an attempt to find out the effect of
Emblica officinalis on hepatoprotection and
biochemical alterations in the liver of Channa
striatus exposed to endosulfan and lindane.
2. Materials and Methods
2.1. Preparation of the Embilica officinalis
extract
Fresh fruits of the E. officinalis obtained
from the local market in Chidambaram,
Tamilnadu, India. These fruits were cleaned in
water and cut into small pieces, air dried,
powdered and extract prepared with double
distilled water by refluxing for 36 hrs. The extract
was redissolved in distilled water just before oral
administration. The extract of E. officinalis fruit
will be called as EEOF.
2.2. Collection of Fish
Channa striatus were collected from the
fish farm located in Puthur, 10 km away from the
Chidambaram, Tamilnadu, India. The collected
fishes were transported in polythene bags filled
half with water, without least disturbance. About
20 fishes were put in each bag and water was well
aerated, using pressurized air from a cylinder. The
fishes brought to the laboratory and disinfected by
0.1% potassium permanganate solution and were
acclimatized in the cement tank (500 L) for three
weeks in well-aerated tap water.

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J. Prakash Sahaya Leon/Life Science Archives (LSA), Volume 1, Issue 2, Page 127 - 137, 2015

129

2.3. Chemical
The physicochemical properties of used chemicals were tabulated in Table - 1.
Properties
Chemical formula
Molecular weight
Physical state at room
temperature
Colour
Vapour pressure at 25C
Flammability
Boiling point
Density at 20/4C

Endosulfan
C9N6Cl6O3Sc
406.93
Crystalline solid
Green to brown
110-6 mm Hg
Non-flammable
106C
1.745 g mL-1

2.4. Experimental Design

In the present investigation, experiment
was performed on adult Channa striatus fish. The
fish were divided into 3 groups (A-C) as follows
Group A Control group
Group B Endosulfan treated group (ES)
Group C Lindane treated group (LI)
The 96 hour LC50 value for endosulfan
and lindane was calculated by standard APHA
method (2005). The 96 hr LC50 value were
obtained as 6.82 g L-1 and 22.65 g L-1 for
endosulfan and lindane respectively. In the present
investigation 1/15th concentration of endosulfan
(0.455 g L-1) and lindane (1.51 g L-1) were used
as exposure concentration. The EEOF aqueous
extract was administered daily by gastric
intubation at 100 mg kg-1 body weight for group A
, B and C for 30 days after exposure periods. The
dose used as 100 mg E. officinalis /kg body weight
for the experiment as immunomodulator and
removal of toxicity is actually a dose below lethal
compared to LD50 value of 1000 mg kg-1 body
weight in fish. The dose used here is sufficient to
carry on their hepatoprotective properties. Liver
tissues were collected from all groups on 10 th,
20th, 30th, 40th, 50th and 60th day of experiment.
2.5. Estimation of Biochemical changes
Protein content in the liver tissue was
estimated by the method of Lowry et al. (1951).

Lindane
C6H6C16
290.83
Crystalline solid
White
9.4 x 10-6 mm Hg
Non-flammable
112.5C
1.85 g mL-1

The tissue (20 mg) was isolated and 2%

homogenate was centrifuged at 3,000 rpm for 15
min. The supernatant was discarded and the
residue was suspended in 1.0 mL of 0.1 N sodium
hydroxide solutions. 0.5 mL of this solution
equivalent to 10 mg of tissue, it was transferred to
a clean test tube and 4 mL of copper carbonate
solution was added. The contents were mixed by
lateral shaking and 0.4 mL of Folin phenol (1:1
dilution) reagent was added. The thoroughly
mixed contents were kept at room temperature for
30 min. The colour developed was read at 600 nm
against a reagent blank in UV visible
spectrophotometer (Jasco Model-650). Bovine
serum albumin (Sigma Chemical Co.) was used to
construct the standard graph. The protein content
in the liver tissue was expressed in mg/g wet
weight of tissue.
Lipids were extracted as described by
Folch et al. (1957), and estimated by the method
of Barnes and Blackstock (1973). 50 mg of liver
tissue was homogenised (5% w/v) in a warring
blender in chloroform-methanol mixture (2:1).
The homogenates were filtered through Whatman
No.1 filter paper, and the residue was
rehomogenised as before and then filtered. The
non-lipid matter from pooled filtrate was removed
by shaking vigorously with 0.88% KCl (added as
one fourth of the volume). 1 mL of filtrate was
taken in a test tube and evaporated under nitrogen
and 1 mL of concentrated H2SO4 was added and
boiled for 10 min. For estimation of total lipid, 0.2
mL of solution was taken and 5 mL of vanillin

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J. Prakash Sahaya Leon/Life Science Archives (LSA), Volume 1, Issue 2, Page 127 - 137, 2015

reagent was added. The developed colour was read

in spectrophotometer at 520 nm against reagent
blank.
2.6. Estimation of Antioxidant enzymes
The activity of superoxide dismutase in
liver tissue was assayed by the method of Kakkar
et al. (1984), based on the formation of NADH
phenazine
meta-sulphate-nitrobluetetrazolium
formazan. The tissue sample was homogenized
with 0.25 M sucrose solution and centrifuged at
1000 rpm in cold condition for 30 min. The
supernatant was dialyzed against Tris-HCl buffer
(0.025 M, pH 7.4). The supernatants obtained
were used as enzyme source. The assay mixture
consisted of 1.2 mL of sodium pyrophosphate
buffer 1.0 mL of appropriately diluted enzyme
preparation and water in a total volume of 3 mL.
Then, the reaction was started by the addition of
0.2 mL of NADH. After incubation at 30C for 90
seconds, the reaction was arrested by the addition
of 1.0 mL of glacial acetic acid. Subsequently, the
reaction mixture was shaken with 4.0 mL nbutanol and allowed to stand for 10 min. Finally,
the butanol layer was separated after
centrifugation and measured at 560 nm. Values
expressed in U/mg protein.
The antioxidant enzyme catalase activity in
liver tissue was assayed by the method of Sinha
(1972) dichromate reacted with acetic acid when
heated in the presence of hydrogen peroxide to
form chronic acetate, which was measured at 620
nm. The enzyme preparation was allowed to split
H2O2 and the reaction was stopped at different
time intervals by the addition of dichromate/acetic
acid reagent. The remaining H2O2 was measured
as chromate acetate at 620 nm. To 1.0 mL of the
phosphate buffer taken in each of four tubes, 0.1
mL of the enzyme preparation was added (tissue
homogenate/RBC lysate). Then, 0.4 mL of H2O2
was added to each tube and the reaction was
stopped at 15, 30, 45 and 60 seconds, by the
addition of 2.0 mL of the dichromate acetic acid
reagent. The tubes were kept at boiling water bath
for 10 min and cooled. Finally, it was read at 620
nm and the rate of removal of H2O2 by catalase
was calculated using hydrogen peroxide standard
in the range of 20-100 mole which were

130

processed similarly along with a blank. Catalase

activity was expressed in U/mg protein.
Reduced glutathione was determined by
the method of Ellman (1959). A known weight of
tissue was homogenized in phosphate buffer (pH
8.0; 0.2 M). 0.5 mL of tissue homogenate was
pipetted out and precipitated with 2.0 mL of 5%
TCA. One mL of the supernatant was taken after
centrifugation and 0.5 mL of Ellmans reagent and
3.0 mL of phosphate buffer were added to it. The
yellow colour developed was read at 412 nm. A
series of standards were treated in a similar
manner along with a blank containing 3.5 mL of
buffer. The amount of glutathione was expressed
g of GSH consumed/min/mg protein.
TBARS in liver tissue was analyzed by the
method of Ohkawa et al. (1979). To 0.2 mL of
tissue homogenate, 0.2 mL of 8.1% sodium
dodecylsulphate and 1.5 mL of 20% acetic acid
were added. The pH of mixture was adjusted to
3.5 with sodium hydroxide, then 1.5 mL of 0.8%
aqueous solution of thiobarbitruic acid (TBARS)
was added to the mixture and the volume was
made up to 4 mL with distilled water. The reaction
mixture was heated in water bath at 95C for 60
min. After cooling in tap water, 1 mL of distilled
water and 5 mL of n-butanol pyridine mixture
were added and shaken vigorously. After
centrifugation at 4000 rpm for 10 min, the organic
layer was removed and absorbance was read at
535 nm. The level of TBARS on tissues was
expressed in moles of TBARS/mg protein.
2.7. Statistical analysis
Results in each group were expressed as
mean S.D. The data were analyzed by an
analysis of variance (ANOVA) followed by
Duncans multiple range test. Significance was
evaluated at P < 0.05.
3. Result
Figure 1 shows the levels of total protein
and total lipid content in liver of control and
experimental fishes.
A significant (P<0.05)
decline in the protein and lipid content were
observed in ES (B) and LI (C) exposure groups
when compared with control.

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J. Prakash Sahaya Leon/Life Science Archives (LSA), Volume 1, Issue 2, Page 127 - 137, 2015

200
150
100
50
0

**

**

10

20

C
*

**

30

A (Control)
*

**

40

**

50

TOTAL LIPID
(mg/g)

TOTAL PROTEIN
(mg/g)

A (Control)

**

25
20
15
10
5
0

**

**

**

* * *** ***
*

10

20

30

40

50

60

60
Exposure

Exposure

131

Treatment

Treatment

DAYS

DAYS

Fig - 1: Effect of EEOF on Total protein and Total lipid content of liver in endosulfan and
lindane treated fish Channa striatus.
*Indicates statistically (P < 0.05) significant difference compared to control group at the same sampling
days.

150
100
50
0

A (Control)

**

**

** *** *** ***

10

20

30

40

Exposure

50

60

SOD
(one unit is as 50%
inhibition of
NBT/ mg protein)

CAT
(moles of H2O2
consumed min/mg
protein)

A (Control)

5
4
3
2
1
0

Treatment

**

**

**

*** *** ***

10

20

30

40

Exposure

DAYS

**

10

20

30

40

Exposure

50

60

Treatment
DAYS

60

Treatment

A (Control)

protein)

**

B C
*
** *** *** **

(moles of TBARS/mg

8
6
4
2
0

50

DAYS

LPO

GSH
(g of GSH consumed/
min/mg protein)

A (Control)

3
2
1
0

**

**

**

**

**

10

20

30

40

50

60

Exposure

Treatment

DAYS

Fig - 2: Effect of EEOF administration on hepatic SOD, CAT, GSH and LPO content in fish
Channa striatus exposed to endosulfan and lindane.
*Indicates statistically (P < 0.05) significant difference compared to control group at the same sampling
days.

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J. Prakash Sahaya Leon/Life Science Archives (LSA), Volume 1, Issue 2, Page 127 - 137, 2015

The administration of E. officinalis in group (A)

shows an elevated level of protein and lipid
content. Group B and Group C treated with E.
officinalis shows recovery effects of protein and
lipid content in liver tissue of fish during the
treatment period (30 days to 60 days).
The antioxidant activities were shown in
Fig. 2. When compared to control, the SOD, CAT
and GSH activity of liver shows a significant
decline in ES and LI exposure groups. The
activities of SOD, CAT and GPx were increased
in group A when treated with E. officinalis. Group
B and Group C treated with E. officinalis shows
recovery effects on anti oxidant activities of liver
tissue in fish during the treatment period (30 days
to 60 days). LPO content in the exposure group B
and C (ES and LI) shows an increased level when
compared to the control group A. The group B,
group C treated with E. officinalis
shows a
decreased level of LPO in all the treatment days.
4. Discussion
Assessment of biochemical parameters;
proteins, carbohydrates and lipids are important to
indicate the susceptibility of organ systems to
pesticide and metal pollutants (Senthilkumaar,
2012). The decreased protein levels in pesticide
stressed tissues strongly suggest the toxicant
induced proteolysis to meet the increased energy
demand (Saravanan et al., 2000; Kumar et al.,
2005). Protein is the most primary biochemical
ingredient present in large quantities in the body
of fish. Protein content rich in the liver than other
organ for various metabolism in fish. In the
present investigation a significant reduction in
protein content after exposure to ES and LI have
been observed. Depletion started from the 10th day
and kept on decreasing till the end of exposure (30
days). The depletion of protein under the stress of
chlorpyrifos toxicity observed in different tissues
of Catla catla, Labeo rohita and Cirrhinus
mrigala indicates the proteolysis, suggesting that
the proteins were utilized to meet the excess
energy demands imposed by the toxic stress. The
protein content declined gradually in gill, liver and
muscle tissues of O. mossambicus when exposed
to deltamethrin and it was reported that it may be
due to the utilization of protein controls to

132

counteract the toxicant stress caused by pesticide

(Rao and Rao, 1979; Rath and Mishra, 1980). The
liver gets affected considerably when there is a
disturbance in protein metabolism. The
accumulation of toxic substance in liver may alter
its function (Premdas and Anderson, 1963). Eva
(1990) reported a continuous reduction in protein
content of the liver when Anabas testudineus was
exposed to sub-lethal concentration of Cuman L.
Reduction in protein content of liver has been
reported in Sarotherodon mossambicus exposed to
lindane (Rajamanickam, 1985). The different
concentrations of malathion, thiodon and ekalux
significantly reduced the total protein in liver of
O. mossambicus (Palanichamy et al., 1986).
Similar observations were noted when the fish
were exposed to pollutants (Lone and Javaid,
1976; Shakoori et al., 1976; Rath and Mishra,
1980; Ramalingam and Ramalingam, 1982). Total
protein content was increased in ES and LI
exposed fishes that were found to recover with of
the treatment with E. officinalis. This might be due
to phytochemical compound present in the natural
products (E. officinalis) in reducing or detoxifying
the effect of ES and LI. The phytochemical
analysis of the E. officinalis fruit revealed the
presence of saponins, tannins, anthraquinones,
coumarins, sterols and/or triterpenes.
These
phytochemical components of the E. officinalis
neutralized the toxic effect of the foreign
substances (Vadivelu and Dawood, 2013).
Commonly increases in the levels of total protein
content in fish are thought to be associated with a
stronger innate immune response (Wiegertjes et
al., 1996). Thus increase in liver total protein in
the present experiment may be an indication to
increased levels of non-specific immunity and
transmit of more humoral compounds into the
blood. Similar results were also observed in C.
carpio fed with feed incorporated with mixed
plant extracts (Inula helenium, Tussilago farfaro,
Brassica nigra, Echinacea purpurea &
Chelidoniume majus) for 60 days and infected
with Aeromonas hydrophila (Sudagar and
Hajibeglou, 2010).
The consumption of fish has been linked to
health benefits, such as reduced risk of coronary
heart disease, which are largely attributable to the

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polyunsaturated fatty acids (PUFA) in fish oils

(Njinkoue et al., 2002) and less amount of lipids.
Lipid content is an essential organic constituent of
the tissues of all animals, and plays a key role in
energy metabolism. Lipids are the best energy
producers of the body next to carbohydrates. The
decreased level of tissue lipid content may be due
to liver dysfunction or mobilization of glycerol or
inhibition of oxidative phosphorylation (Chezhian
et al., 2010). Gilbert and OConnor (1970)
reported that lipids are vital to embryogenesis,
providing two third of energy by oxidation. In the
present investigation, there was a decrease in the
total lipid content of liver in C. striatus exposed to
ES and LI. The lipid content decreased in the liver
tissues during the exposed period for both the
toxicants (ES and LI) justifying the utilization of
energy depot to meet the requirement of more
energy for detoxification process and also to
balance the hindrance of normal metabolism
(Senthilkumaar, 2012). Also the investigation
shows the recovery effect during EEOF treatment
both ES and LI group (30 days to 60 days).
Oxidative stress which is characterized by
an imbalance in reactive oxygen species (ROS)
and antioxidative defense has been demonstrated
to play a critical role in several neurodegenerative
diseases (Lin and Beal, 2006; Zhoua et al., 2013).
Reactive oxygen species (ROS) are part of the
normal oxidative metabolism, but when produced
in excess, they cause tissue injury (Nurulain et al.,
2013).
Damage induced by ROS includes
alterations of cellular macromolecules such as
membrane lipids and proteins. The damage may
alter cell function through changes in intracellular
calcium or intracellular pH and eventually can
lead to cell death. (Kehrer et al., 1990; Swann et
al., 1991). Specially adapted systems normally
counteract the damaging effects of ROS by
producing antioxidant enzymes like Superoxide
Dismutase (SOD), Catalase (CAT), Glutathione
peroxidase (GPx) and Glutathione reductase
(GSH) and free radical scavengers (Vit C and E)
remove ROS and protect the organisms from
oxidative stress. SOD metabolizes superoxide
anion (O2-) into molecular oxygen and hydrogen
peroxide (H2O2). The H2O2 is detoxified into
water (H2O) and oxygen (O2) by peroxysomal

133

Catalase (CAT). This reaction diminishes the

destructive oxidative processes in cells. The
levels of antioxidant enzyme have been
extensively used as an early warning indicator of
pollution (Lin et al., 2001; Naveed and Janaiah,
2011). In the present investigation, a significant
decrease in the SOD, CAT and GSH activities
whereas increases in the lipid peroxidation (LPO)
levels were recorded in liver of C. striatus
exposed to ES and LI for the exposure periods.
The decreased SOD activity could be due to the
oxidative inactivation of the enzyme as a result of
excessive reactive oxygen species generation
(Pigeolot et al., 1990). The decrease in CAT
activity could be due to its inactivation by
superoxide radical or due to decrease in the rate of
the reaction as a result of the excess production of
H2O2 (Obaiah and Usha, 2012). The present study
agrees with Crupkin et al. (2013) which decrease
the SOD, CAT and GSH of Australoheros facetus
when exposed to endosulfan. The activity of SOD
and CAT were observed in liver, brain and kidney
of
Channa
punctatus
during
sublethal
concentration of triazophos for different exposure
periods. The SOD and CAT activity significantly
decreased in all the tissues during the toxic
exposure periods (Naveed and Janaiah, 2011).
The GSH/GST detoxification system is an
important part of cellular defence against a large
array of injurious agents. GSH offers protection
against oxygen derived free radicals and cellular
lethality following exposure to radiation (Biaglow
and Vames, 1987). In normal condition, the
inherent defense system including glutathione and
antioxidant enzymes protects against the oxidative
damage. GSH is versatile protector and executes
its protective function through free radical
scavenging restoration of the damage molecule by
hydrogen donation, reduction of peroxides and
maintenance of protein thiols in the damage state
(Bump and Brown, 1990). The present study
demonstrates a significant reduction in liver GSH
following pesticide exposure (Group B and C).
This could be due to the enhanced utilization of
the antioxidant system as an attempt to detoxify
the free radicals generated by exposure.
Administration of EEOF increases the GSH level
when it was administrated after the endosulfan and

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J. Prakash Sahaya Leon/Life Science Archives (LSA), Volume 1, Issue 2, Page 127 - 137, 2015

lindane exposure. The higher availability of GSH

increases the ability to cope up with free radicals
produced by pesticide. The increased GSH level
in the present study that protection given by E.
officinalis may be mediated through the
modulation of cellular antioxidant levels.
The results shown in the Fig. 2, it clearly
indicates a significant increase of lipid
peroxidation (LPO) in the liver exposed to ES and
LI of fish C. striatus as compared to the control.
Lipid peroxidation and oxidative damage is
regarded as one of the primary causes of cellular
damage. It is presume that ES and LI increases
cell permeability to Ca2+ and increases cellular
concentration of Ca2+. Also, in the presence of
prooxidants, ES and LI uncouple oxidative
phosphorylation. This results in an increased
leakage of electron from the respiratory chain,
thus producing oxygen and form H2O2. An
increase in lipid peroxidation accompanied by the
leakage of Ca2+ from Ca2+ loaded microsomes
would produce further damage to the endoplasmic
membrane (Verma and Mathew, 1998). Previous
studies have shown increased lipid peroxidation in
liver of Heteropenustes fossilis which would
suggest changing pro-oxidant processes and a
need for antioxidant responses to protect against
increased ROS production (Parihar et al., 1996).
Liet et al. (2003) found that 3, 4, dichloroaniline, a
chemical intermediate in the synthesis of
herbicides has significantly induced the increase
of MDA content in the liver of Carassius auratus.
The fish, Anguilla anguilla exposed to polluted
water induces the tissue specific peroxidative
damage in gill, kidney and liver (Ahmad et al.,
2004). Uner et al. (2001) studied the effects of
cypermethrin on antioxidant enzyme activities and
lipid peroxidation in liver and kidney of the
freshwater fish, Oreochromis niloticus and
Cyprinus carpio. The overall effect of lipid
peroxidation is to decrease membrane fluidity and
increase leakiness of the membrane leading to
complete loss of membrane integrity (Halliwell
and Gutteridge, 1985).
The biochemical and antioxidant activities
were recovered in the ES and LI treated groups
(30 days to 60 days), it shows the presence of
phytochemical component in the E. officinalis.

134

Extract of E. officinalis has been shown to have

antioxidant and antiperoxidant properties due to
the presence of low molecular weight tannoids,
mainly emblicanin-A (37%), emblicanin-B (33%),
punigluconin (12%), pedunculogin (14%) and
galic acid (Bhattacharya et al., 1993). The in vitro
antioxidant activity of tannoids was demonstrated
as well (Ghosal et al., 1996). Some of the plants
like Glycyrrhiza glabra, Rubia cordifolia and
Phylanthus emblica have also been reported to
possess antioxidant and free radical scavenging
activities (Tripathi et al., 1997; Korina and
Afanasav, 1997)). The emblicanins are likely to
the major antioxidant principles, not only because
they are the major constituents of E. officinalis but
also because of their reported antioxidant actions
in vitro (Ghosal et al., 1996) and in vivo
(Bhattacharya et al., 1993, 2000).
5. Conclusion
From the present study we concluded that
endosulfan and lindane are responsible for
irreversible damages in the liver as well as
decreased biochemical properties and antioxidant
activity simultaneously significantly increase in
lipid peroxidation level which lead to unhealthy
condition of fish. Our results revealed that the
endosulfan and lindane induced oxidative stress
being harmful to the fish. Based on the findings it
can be concluded that the extract of Emblica
officinalis have cytoprotective properties on liver
damage. Extract of Emblica officinalis contain
antioxidants and several flavonoids, these reduces
the oxidative stress and protect against the tissue
damage. E. officinalis neutralizes the oxidizing
potentials of ROS induced by endosulfan and
lindane, these activities maintain membrane
integrity and viability.
Acknowledgement
The first author is indebted to University
Grant Commission, New Delhi, India for the
financial assistance for conducting this research
work (Minor research grant F. No. 40-501/2011
(SR) ) and also thankful to the authorities of
Annamalai University, Annamalainagar for
providing all facilities to carry out the work.

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J. Prakash Sahaya Leon/Life Science Archives (LSA), Volume 1, Issue 2, Page 127 - 137, 2015

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