Food and Chemical Toxicology 49 (2011) 991–998

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Food and Chemical Toxicology

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Protective effect of gallic acid against lindane induced toxicity in experimental rats
V. Vijaya Padma ⇑, P. Sowmya, T. Arun Felix, R. Baskaran, P. Poornima
Department of Biotechnology, School of Biotechnology and Genetic Engineering, Bharathiar University, Coimbatore 641046, Tamil Nadu, India

a r t i c l e i n f o a b s t r a c t

Article history: Lindane is an organochlorine pesticide that persists in the environment, bioaccumulate through food
Received 15 July 2010 chain and has a risk of causing adverse effects to human health and the environment. It induces cell dam-
Accepted 4 January 2011 age by producing free radicals and reactive oxygen species. The aim of the present study is to investigate
Available online 8 January 2011
the protective effect of gallic acid (a plant derived polyphenol) against lindane induced hepatic and renal
toxicity in rats. Liver damage was assessed by hepatic serum marker enzymes like SGOT, SGPT and ALP
Keywords: and histopathological observation. Renal damage was observed by histopathological examination and
Lindane
serum markers like creatinine and urea. Treatment with lindane increased the levels of lipid peroxidation,
Gallic acid
Renal markers
serum marker enzyme activity with a concomitant decrease in GSH, CAT, SOD, GPx and GST. Histological
Liver markers alterations were also observed in kidney and liver tissue with lindane treatment. Co-treatment of gallic
Histopathology acid significantly prevented the lindane induced alterations in kidney and liver tissues with a decrease in
Antioxidant system LPO, serum marker enzyme activity and a significant increase in antioxidant levels. These results suggest
that gallic acid has protective effect over lindane induced oxidative damage in rat liver and kidney.
Ó 2011 Elsevier Ltd. All rights reserved.

1. Introduction
Organochlorine pesticides are organic compounds that persist
in the environment, bioaccumulate through the food chain and
have a risk of causing adverse effects to human health and environ-
ment. These pesticides are characterized by their cyclic structure,
number of chlorine atoms and low volatility, and can be divided
into four groups: dichlorodiphenylethanes (such as DDT), cyclodi-
enes (such as dieldrin, endosulfan, and heptachlor), chlorinated
benzenes (such as hexachlorobenzene) and cyclohexanes (such as
lindane) (Ledirac et al., 2005). Lindane, a c-isomer of hexachlorocy-
clohexane has largely been used as an organochlorine pesticide
and is widely spread in the environment due to its long lifetime
(Wauchope et al., 1992). It has been used widely as therapeutic
scabicide, pediculicide and ectoparasiticide (Fidan et al., 2008). Lin-
dane is also presently used in lotions, creams and shampoos for the
control of lice and mites in humans (Safe, 1993; Budavari et al.,
Abbreviations: ALP, alkaline phosphatase; ANOVA, analysis of variance; CAT,
catalase; CPCSEA, Committee for the Purpose of Control and Supervision of
Experiments and Animals; DDT, dichlorodiphenyltrichloroethane; GPx, glutathi-
one peroxidase; GSH, reduced glutathione; GST, glutathione-s-transferase; H&E,
hematoxylin and eosin; LDH, lactate dehydrogenase; LPO, lipid peroxidation; ROS,
reactive oxygen species; SGOT, serum glutamate oxaloacetate transaminase; SGPT,
serum glutamate pyruvate transaminase; SOD, superoxide dismutase; TBARS, thio
barbituric acid reactive substances.
⇑ Corresponding author. Tel.: +91 9894173494; fax: +91 422 2422387.
E-mail address: padma.vijaya@gmail.com (V. Vijaya Padma).
1989). Compounds of this chemical class have very low water sol-
ubility but are highly soluble in lipids and bioaccumulate (Murphy,
1986). They adsorb soil particles and occur in soil of high organic
content (Menzer and Nelson, 2000). Toxic effects of lindane in
mammals include convulsions, ataxia, prostration, damage to fatty
tissues and inhibition of sperm motility in sea urchins (Nelson,
1990; Murphy, 1986).
Normal cellular function depends on a balance between reac-
tive oxygen species (ROS) produced and antioxidant defense mech-
anisms present in the cell. ROS arise as by-products of normal
cellular metabolism or may be the consequence of exposure to cer-
tain chemicals (Kerr et al., 1996; Moslen, 1994). Reactive species
derived from chemicals, oxygen or nitrogen has been implicated
as putative noxious intermediates responsible for cellular damage.
Because electrophilic metabolites or radicals can readily interact
with essential biomolecules or bind covalently to cellular compo-
nents which leads to structural and functional alterations in cells
(Fernandez et al., 2003; Comporti, 1989; Kappus, 1987). Pesticide
chemicals may induce oxidative stress leading to generation of free
radicals and alterations in antioxidants and scavengers of oxygen
free radicals (Banerjee et al., 1999). Lindane has been reported to
induce oxidative stress by interacting with the cell membrane,
triggering the generation of ROS and altering the level of antioxi-
dant molecules which in turn cause functional impairment in
blood brain barrier and alteration in cytochrome P450 mono oxy-
genase enzymes thus lindane causes severe physiological dysfunc-
tion in various organ systems (Barros et al., 1991; Bano and Bhatt,
2007).

0278-6915/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.fct.2011.01.005
992 V. Vijaya Padma et al. / Food and Chemical Toxicology 49 (2011) 991–998
The development of oxidative stress in liver and kidney by lin-
dane intoxication has been considered as a possible toxic mecha-
nism of the insecticide. Lindane is biotransformed by liver
microsomal enzymes to a variety of metabolites, including poly-
chlorocyclohexenes that are susceptible to GSH conjugation for
elimination (Videla et al., 1990). In addition, induction of the liver
microsomal cytochrome P450 system is also observed, with an
enhancement in the rate of free radical generation and in the
magnitude of lipid peroxidation indicators (Junqueira et al.,
1988; Videla et al., 1989; Bainy et al., 1993). Concomitantly, lin-
dane alters some antioxidant mechanisms of the cells in kidney,
including decrease in superoxide dismutase and catalase activi-
ties. The changes in these biochemical parameters related to oxi-
dative stress are dose and time dependent, and coincide with the
onset and progression of morphological lesions (Junqueira et al.,
1986). Due to the extensive use of lindane for public health pur-
poses in developing countries and the incidence of toxicity in hu-
mans upon its exposure, it is considered important to investigate
whether lindane induced hepato and nephrotoxicity could be
modulated by an antioxidant which could reduce the oxidative
stress.
Phenolics have been reported to have a capacity to scavenge
free radicals. The antioxidant activity of phenolics is mainly due
to their redox properties, which allow them to act as reducing
agents, hydrogen donators and singlet oxygen quenchers (Kahko-
nen et al., 1999). Gallic acid as well as its derivatives has impor-
tant antioxidant activities by displaying radical scavenging
abilities. They are polyphenols which scavenge free radicals by
donating hydrogen to radical while it oxidizes to quinone which
is stable. Other than stabilizing radicals which are products of cell
metabolism, phenolic compounds can also combine with copper
and iron metal ions, which can induce the radical production
via Fenton reaction. Hynes (2001) showed that complex formation
between ferric ions and gallic acid can result in the oxidation of
gallic acid to hydroxyquinone and reduction of ferric ions to fer-
rous ions.
Gallic acid is described as an excellent free radical scavenger
and as an inducer of differentiation and programmed cell death
in a number of tumor cell lines (Salucci et al., 2002; Inoue et al.,
1994; Kawada et al., 2001). Recently, it is shown that gallic acid
from rose flowers exhibits antioxidant effects in senescence accel-
erated mice and can reinstate the activities of catalase and gluta-
thione peroxidase (Li et al., 2005).
In the present study, we investigated the potential protective
effects of gallic acid in lindane induced liver and kidney damage
in Wistar rats.
2. Materials and methods
2.1. Chemicals
Lindane (c-isomer) and gallic acid (98.5%) were purchased from Sigma Aldrich
Pvt. Ltd., Bangalore, India and were dissolved in olive oil (100%) and water, respec-
tively. All other chemicals were purchased from HiMedia laboratories, Mumbai,
India.
2.2. Animals
Female albino Wistar rats weighing between 150–200 g each were used for this
experiment. They were procured from Kerala Agricultural University Mannuthy,
Kerala, India. The rats were maintained in a controlled environment under standard
conditions of temperature (28 ± 2 °C) and humidity with an alternating light and
dark cycle. The animals were fed with commercially available pelleted rat chow
(Sai Durga private limited, Bangalore, India) and water ad libitum. After a week of
acclimatization, rats were divided into control and test groups. Six rats were used
in each treatment group. The study protocol was approved from the Institutional
Animal Ethics Committee constituted in accordance with the rules and guidelines
of the CPCSEA (Committee for the Purpose of Control and Supervision of Experi-
ments and Animals), India.
2.3. Treatment schedule
Lindane (100 mg/kg) dissolved in 0.3 ml of olive oil and gallic acid (50 mg/kg)
dissolved in 0.3 ml water, were administered to rats orally for 30 days by oral intu-
bation method.
2.4. Experimental procedure
The animals were divided into five groups with 6 rats in each group. Group I
served as control. Group II served as vehicle control, treated with olive oil (Vehicle
for lindane). Group III received lindane alone (100 mg/kg body weight). Group IV
was treated with gallic acid alone (50 mg/kg body weight). Group V was co-treated
with lindane (100 mg/kg body weight) and gallic acid (50 mg/kg body weight). First
the animals were treated with lindane followed by gallic acid administration with
about 15 min duration between the treatments.
After 30 days of treatment period, the animals were deprived of food over-
night and anesthetized by exposing to diethyl ether and then sacrificed by cervical
decapitation. Blood was collected from jugular vein and serum was separated and
used for liver and kidney marker assays. The liver and kidney tissues were dis-
sected out, washed in ice-cold saline, patted dry and weighed. A small portion
of the liver and kidney tissue were stored in 10% formalin for histopathological
examination. From the remaining tissue, about 100 mg tissue from liver and
kidney were weighed and homogenized in chilled 0.1 M Tris–HCl buffer in
Potter–Elvehjem Teflon homogenizer. The homogenates were used for biochemi-
cal investigation.
2.5. Histopathology
A small portion of the liver and kidney tissue from the control and experimental
animals was fixed in 10% neutral buffered formalin and processed by standard pro-
cedure for paraffin embedding and serial sections of about 5 lm size were cut and
were stained with hematoxylin and eosin (H&E) dyes.
2.6. Analytical procedure
The serum was used for the estimation of protein (Lowry et al., 1951), urea
(Natelson et al., 1951), creatinine (Broad and Sirota, 1948) and hepatic marker en-
zymes namely serum glutamate oxaloacetate transaminase (SGOT), serum gluta-
mate pyruvate transaminase (SGPT) (Reitman and Frankel, 1957), alkaline
phosphatase (ALP) (Anderch and Szczypinski, 1947) and lactate dehydrogenase
(LDH) (King, 1965). The liver and kidney tissue homogenates were used for the as-
say of protein (Lowry et al., 1951), thio barbituric acid reactive substances (TBARS)
(Ohkawa et al., 1979), superoxide dismutase (SOD) (Marklund and Marklund,
1974), catalase (CAT) (Takahara et al., 1960), glutathione peroxidase (GPx) (Rotruck
et al., 1973), glutathione-s-transferase (GST) (Habig et al., 1974) and reduced gluta-
thione (GSH) (Moron et al., 1979).
2.7. Statistical analysis
Biochemical data were analyzed using one-way analysis of variance (ANOVA)
followed by Tukey’s multiple comparison test.
3. Results
3.1. Histopathology of rat liver
Fig. 1 demonstrated that the rats treated with lindane alone
causes extensive liver injuries characterized by vacuolar degenera-
tion of hepatocytes and massive degradation of central vein. How-
ever, the animals treated with both lindane and gallic acid showed
reduction in liver damage with slight damage in central vein. Con-
trol animals and animals treated with gallic acid alone showed
normal architecture.
3.2. Histopathology of rat kidney
Fig. 2 demonstrated that the rats treated with lindane showed
degeneration and shrinkage of glomerulus and degeneration of
proximal and distal tubules. Lindane and gallic acid co-treatment
significantly reduced the degenerative changes in renal tissue of
rats when compared to the animals treated with lindane alone.
Whereas control animals and animals treated with gallic acid alone
showed normal architecture.
 V. Vijaya Padma et al. / Food and Chemical Toxicology 49 (2011) 991–998 993

Fig. 1. Histopathological changes of liver tissue. (A) Liver of control rats showing normal architecture H&E 400. (B) Liver of lindane treated rats H&E 400. (C) Liver of gallic
acid treated rats showing apparently normal architecture H&E 400. (D) Liver of rats treated with lindane followed by gallic acid H&E 400.

Fig. 2. Histopathological changes of kidney tissue. (A) Kidney of control rats showing normal architecture H&E 400. (B) Kidney of lindane treated rats H&E 400. (C) Kidney
of gallic acid treated rats showing apparently normal architecture H&E 400. (D) Kidney of mis treated with lindane followed by gallic acid H&E 400.

3.3. Biochemical parameters
Several enzymes in liver tissue have long been considered as
effective biochemical markers to understand the early injury. In
the present study, lindane treated rats showed increased activity
of transaminases (SGOT & SGPT), alkaline phosphatase (ALP) and
Lactate dehydrogenase (LDH). Gallic acid restored physiological
integrity of hepatocytes, thereby reducing the elevated values
of serum SGOT, SGPT, ALP and LDH in groups treated with lin-
dane and gallic acid (Fig. 3). Kidney markers like creatinine and
urea were estimated in the serum of experimental animals to
confirm the renal damage. Levels of creatinine and urea were
increased in lindane treated groups. Whereas in gallic acid and
lindane co-treated groups, the level of urea and creatinine were
significantly decreased (Fig. 3). The hepatic and renal marker
levels in the animals treated with gallic acid or olive oil alone
were found to be non-significant when compared to control
animals.
994 V. Vijaya Padma et al. / Food and Chemical Toxicology 49 (2011) 991–998

Fig. 3. Effect of gallic on hepatic and rental markers in lindane treated rats Group 1 – Control, Group II – Olive oil (vehicle for lindane), Group III – lindane, Group 1V – gallic
acid, Group V – lindane + gallic acid. Each bar represents, mean ± standard error of mean (SEM) of each group. ⁄⁄⁄P < 0.001, compared to control. +++P < 0.001, compared to
lindane treated group. NS, not significant (#); compared to control (One way ANOVA followed by Tukey’s multiple comparison.

As presented in Table 1, the protein level in both serum and tis-
sue of lindane treated rats were found to be lower compared to
control animals. Whereas animals which received both lindane
and gallic acid as co-treatment have showed significantly in-
creased serum and tissue protein compared to lindane treated rats.
GSH content was significantly decreased in animals treated with
lindane alone. GSH content in animals administered with both lin-
dane and gallic acid was found to be significantly increased when
compared to lindane treated animals. Liver and kidney lipid perox-
idation (LPO) contents were significantly increased in the rats trea-
ted with lindane alone. Rats treated with both lindane and gallic
acid significantly reduced the LPO products in liver and kidney,
compared with the animals treated with lindane alone. However,
the animals which were treated with gallic acid alone and olive
oil (vehicle for lindane) showed no significant difference in the val-
ues of protein, LPO and GSH when compared to control animals.
 V. Vijaya Padma et al. / Food and Chemical Toxicology 49 (2011) 991–998 995

Table 1
Effect of gallic acid on lindane induced alterations in the lipid peroxidation and antioxidant system.

S. No Parameters Group I Group II Group III Group IV Group V

NS NS
1 Protein Serum 15.29 ± 1.13 15.19 ± 1.42 8.93 ± 0.67⁄⁄⁄
14.87 ± 1.81 12.27 ± 0.76+
Liver 19.23 ± 3.26 19.12 ± 1.38NS 9.28 ± 1.26⁄⁄⁄ 19.27 ± 1.56NS 14.98 ± 1.89++
Kidney 48.31 ± 1.72 47.40 ± 0.71NS 33.47 ± 1.22⁄⁄⁄ 47.42 ± 2.43NS 41.16 ± 1.42+++
2 LPO Liver 14.71 ± 0.36 14.39 ± 0.29NS 22.82 ± 2.14⁄⁄⁄ 14.65 ± 0.21NS 15.06 ± 1.24+++
Kidney 36.89 ± 1.15 38.43 ± 1.29NS 79.78 ± 0.93⁄⁄⁄ 37.67 ± 1.08NS 53.83 ± 0.97+++
3 GSH Liver 26.93 ± 0.34 26.31 ± 0.55NS 13.39 ± 0.13⁄⁄⁄ 26.82 ± 0.14NS 24.42 ± 0.02+++
Kidney 37.37 ± 1.40 36.49 ± 0.81NS 25.57 ± 1.30⁄⁄⁄ 36.15 ± 1.40NS 30.99 ± 1.17+++
4 SOD Liver 0.44 ± 0.04 0.41 ± 0.17NS 0.26 ± 0.12⁄⁄⁄ 0.45 ± 0.16NS 0.32 ± 0.12+++
Kidney 0.82 ± 0.03 0.83 ± 0.03NS 0.51 ± 0.02⁄⁄⁄ 0.82 ± 0.04NS 0.71 ± 0.02+++
5 CAT Liver 2.69 ± 0.39 2.58 ± 0.60NS 1.22 ± 0.70⁄⁄⁄ 2.48 ± 0.29NS 2.01 ± 0.63 ++
Kidney 0.32 ± 0.02 0.31 ± 0.03NS 0.18 ± 0.01⁄⁄⁄ 0.31 ± 0.03NS 0.26 ± 0.02++
6 GST Liver 6.33 ± 0.05 6.32 ± 0.12NS 4.22 ± 0.02⁄⁄⁄ 6.39 ± 0.13NS 5.76 ± 0.11+++
Kidney 5.60 ± 0.17 5.70 ± 0.23NS 3.27 ± 0.19⁄⁄⁄ 5.63 ± 0.31NS 4.55 ± 0.16+++
7 GPx Liver 52.19 ± 1.19 52.01 ± 1.46NS 37.96 ± 2.0⁄⁄⁄ 51.91 ± 1.89NS 44.39 ± 1.66+
Kidney 46.84 ± 1.09 45.89 ± 1.07NS 35.04 ± 1.94⁄⁄⁄ 45.73 ± 2.93NS 41.12 ± 1.57+++

Group I – Control, Group II – Vehicle control, Group III – Lindane (100 mg/kg Body weight), Group IV – gallic acid (50 mg/kg Body weight), Group V – Lindane (100 mg/kg Body
weight) and gallic acid (50 mg/kg Body weight) co-treatment.
Values are means ± standard error of mean of each group. ⁄⁄⁄P < 0.001, compared to control. +P < 0.05, ++P < 0.01, +++P < 0.001, compared to lindane treated group. NS, not
significant, compared to control. (One way ANOVA followed by Tukey’s multiple comparison).
Units are expressed as: protein in serum as mg/dl and in tissue as mg/g of tissue, LPO in nmoles of TBA reactants/mg protein, GSH activity in n moles/g of tissue, CAT, SOD, GST
and GPx activities in Units/mg protein.

CAT, SOD, GST and GPx activities were reduced significantly in
lindane treated animals compared to control. Whereas in lindane
and gallic acid co-treated animals, significant increase in activity
of these enzymes were observed compared to lindane treated
group. The animals which were treated with gallic acid alone and
olive oil alone (vehicle for lindane) showed no significant differ-
ence in the antioxidant enzyme activity when compared to control
animals (Table 1).
4. Discussion
In recent years, there is considerable interest in free radical
mediated damage in biological systems due to pesticide exposure
(Banerjee et al., 1999, 2001) and the search for herbal drugs with
antioxidant activity has gained importance as the dietary intake
of antioxidants obtained from natural sources is considered to be
relatively safe and without undesirable side effects (Xavier et al.,
2004). Plant polyphenols are well known to show biological activ-
ity, such as antimutagenicity, anticarcinogenicity and antioxidative
activity. Gallic acid, a naturally occurring plant phenol, was also
found to be a strong antioxidant and exhibits antimutagenicity
(Madsen and Bertelsen, 1995; Nakatani, 1992). Gallic acid is wide-
spread in plant foods and beverages such as tea and wine and was
proven to be one of the anticarcinogenic polyphenols present in
green tea (Kerry and Abbey, 1997; Caccetta et al., 2001). Gallic acid
is a strong chelating agent and forms complexes of high stability
with iron (III) (Sroka et al., 1994; Li et al., 2000). Antioxidant capac-
ity of gallate esters against hydroxyl, azide and superoxide radicals
has also been reported (Masaki et al., 1995; Satoh et al., 1998; Bors
and Michel, 1999; Metelitza et al., 2001).
Lindane is a toxic compound via oral exposure, with a reported
oral LD50 of 88–190 mg/kg in rats (Smith, 1991). Chronic exposure
of lindane leads to bioaccumulation in the fatty tissues of animals
but our study period is limited to 1 month, we have chosen lindane
dose from the previous reports. The 30 days may serve as subacute
period for the lindane toxicity so that this may reflect the toxicity
through the contaminated environment and hence its bioaccumu-
lation period. In 2006, Etim et al. have reported the effect of aloe
vera extract on the lindane induced toxicity, in this study they
administered lindane at a concentration of about 100 mg/kg of
body weight for 4 weeks. The dose of gallic acid was chosen from
the previous studies, in 2010, Giftson et al. showed the chemopre-
ventive efficacy of gallic acid at a concentration of (50 mg/kg/day)
against 1,2-dimethyl hydrazine induced rat colon carcinogenesis.
To test the protective effect of gallic acid against carbon tetrachlo-
ride-induced chronic liver injury in rats, Tung et al. (2009) used the
gallic acid at a concentration of 50 mg/kg bodyweight. As many of
the researchers used the same concentrations for gallic acid we
chose this concentration to analyze its protective effect on lindane
induced toxicity.
Histopathological studies revealed several abnormalities in the
liver and kidney tissue of lindane treated animals. Compared with
control, lindane treated animal showed megalocytosis in hepato-
cytes, vacuolar degeneration, sinusoidal as well as venous conges-
tion and periportal lymphocytic infiltrations in liver. Degeneration
of glomerulus, proximal tubule and distal tubule were observed in
kidney of lindane treated rats. Suter (1983) observed liver and kid-
ney effects in rats fed with lindane showed centrilobular hypertro-
phy and necrosis, tubular distension and basophilic tubules,
respectively. In gallic acid and lindane co-treated animals only
slight degeneration of cells was found.
One of the most sensitive and dramatic indicators of hepatocyte
injury is the release of intracellular enzymes, such as transami-
nases and serum alkaline phosphatase in the circulation after lin-
dane administration. The elevated activities of these enzymes are
indicative of cellular leakage and loss of the functional integrity
of the cell membranes in liver (Rajesh and Latha, 2004). In the pres-
ent study, lindane treated rats showed significant increase in the
activity of transaminases and alkaline phosphatase compared to
control rats. This result is consistent with the earlier report by Etim
et al. (2006). Gallic acid restored physiological integrity of hepato-
cytes thereby reducing the values of SGOT, SGPT and ALP in serum.
This suggests that antioxidant and free radical scavenging proper-
ties, reported previously by Salah et al. (1995) and Kawashima
et al. (1996), may account for the hepatoprotective effect of gallic
acid.
Lactate dehydrogenase is a well known marker for cell damage.
When cells are damaged due to oxidative stress or destroyed due
to deficient oxygen supply or glucose, the membrane becomes per-
meable or may rupture which results in the leakage of this enzyme.
This enzyme enters into the blood stream thus increasing their
concentration in the serum (Mathew et al., 1985). In the present
study LDH leakage is significantly increased in the serum of rats
exposed to lindane. Gallic acid administration to lindane treated
rats significantly decreased LDH level in serum compared to rats
996 V. Vijaya Padma et al. / Food and Chemical Toxicology 49 (2011) 991–998

treated with lindane alone. Kocak et al. (1992) reported the protec- property of gallic acid. Our results with gallic acid are in line with
tive effect of gallic acid pretreatment against isoproterenol induced previously published reports of Hatano et al. (1989) and Son and
cardiotoxicity in Wistar rats. Lewis (2002).
Kidney markers like creatinine and urea were also estimated in GSH an important non-protein thiol, which in conjugation with
the serum of experimental animals to confirm the renal damage. GPx plays a significant role in protecting cells against cytotoxic and
The amount of creatinine excreted each day is constant and can carcinogenic chemicals by scavenging ROS (Micheiels et al., 1994).
be used as an indicator of the normalcy of the excretory function This tripeptide plays a vital role with GPx in the detoxification of
of the kidneys. High blood creatinine levels could indicate an many environmental carcinogens and free radicals (Meister,
inability of the kidney to excrete creatinine, resulting from renal 1974). In the present study GPx level in the liver and kidney were
disease (Marks and Lieberman, 2009). So any damage to kidney decreased in rats treated with lindane alone when compared to
will show an improper excretion of creatinine. In lindane treated control animals. Gallic acid supplementation to the lindane treated
rats, elevated level of serum creatinine was observed than control. rats increased the levels of GPx. An increase in intracellular GPx
But in rats co-treated with lindane and gallic acid, the level of cre- concentration induced by gallic acid has been previously reported
atinine was significantly decreased. by Nerya et al. (2004).
Urea is an end product of protein and amino acid metabolism. GST is a potent antioxidant that provides cells with a substan-
Kidney filters excess urea into the urine and in sweat, but some tial degree of protection against oxidative stress. In the present
goes into the bloodstream as serum urea. It is a well known fact study lindane significantly decreased GST activity in liver and kid-
that, if blood urea levels are low, a problem centered in the liver ney tissues. Spychala (2000) showed a significant decrease in GST
might be suspected because urea is produced in the liver. Con- in male mice when treated with lindane.
versely, high blood levels of urea suggest that the kidney is not In our study, gallic acid administration to lindane treated rats
excreting urea normally (Vanholder et al., 1992). From our results showed elevated level of GST, compared to lindane treated rats.
it is clear that, lindane treated animals showed high level of urea Studies conducted by Hsu and Yen (2007) reported significant
than control animals. But in gallic acid and lindane co-treated ani- increase in GST in the hepatic tissue of rats with high fat diet-
mals the level of urea was significantly decreased, which could be induced obesity. This may be due to the strong antioxidant
due to the protective effect of gallic acid. property of gallic acid (Gali et al., 1992). GST activity was also
In the present study, lindane caused an increase in LPO level in significantly increased in kidney tissues of gallic acid co-treated
liver and kidney tissues which is an indicator of oxidative stress. animals, than the rats treated with lindane alone. Similar result
LPO products are measured as index of oxygen free radical. The of GST activation upon gallic acid administration in rats was also
present result of increased LPO level after lindane exposure indi- reported by Shinno et al. (2005). Phenolic hydroxyl groups are
cates an increased oxygen free radical generation by pesticides. known to be potent in scavenging free radicals and the OH group
This result is in accordance with previous studies using lindane at the para position to the carboxylic group is especially effectual
and other pesticides (Koner et al., 1998 and Banerjee et al., for the antioxidant activity (Lu et al., 2006). Thus the three hydro-
2001). Induction of cytochrome P450 and other microsomal en- xyl groups present in gallic acid may be responsible for its
zyme by various pesticides, e.g. carbamate, has been reported antioxidant property.
and it is possible that lindane mediated free radical generation
could be through induction of these enzyme (Hayes, 1982 and
Puatanochokchai et al., 2006). Gallic acid administration to lindane
treated rats resulted in a significant decrease in LPO level com-
pared to rats treated with lindane alone. These results are similar
to the observation of another study where gallic acid was shown
to decrease LPO level in carbon tetrachloride induced damage in al-
bino rats (Jadon et al., 2007).
GSH is an important molecule in the cellular defense against
oxidative stress. In its reduced form, it is necessary for the detoxi-
fication of xenobiotics. In the present study, exposure of animals to
lindane caused a significant decrease in GSH levels in liver and kid-
ney tissues. Various pesticides including lindane have been shown
to decrease GSH level. This reduction in GSH level could be due to
direct conjugation of GSH with electrophiles whose increased pro-
duction may result from pesticide exposure or could be due to
inhibition of enzymes, like glutathione reductase, glutathione per-
oxidase, glucose-6-phosphate dehydrogenase, etc. which are in-
volved in GSH synthesis and regeneration (Reed, 1990). Gallic
acid administration produced an increase in the level of GSH in
both liver and kidney tissues and also attenuated the decrease in
GSH induced by lindane.
Superoxide dismutase (SOD) and Catalase (CAT) are two impor-
tant enzymic antioxidants that act against toxic oxygen free radi-
cals such as superoxide (O2) and hydroxyl ions (OH) in
biological systems (Burton et al., 1983). CAT prevents oxidative
hazards by catalyzing the formation of H2O and O2 from H2O2
(Kumar and Kuttan, 2003). In our present study we observed a de-
crease in the SOD and CAT activities of the liver and kidney tissues There were reports for the increased expression of mRNA for
on lindane administration. Gallic acid administration to lindane CYP1A1, 1A2, 2B1, 2B2, and 2E1 as well as associated catalytic
treated rats significantly increased the SOD and CAT activities. activities in rats treated with lindane (Johri et al., 2008). Gallic acid
This could be due to the free radical scavenging and antioxidant has been reported to decrease the CYP2E1 levels in the CCl4 treated
 V. Vijaya Padma et al. / Food and Chemical Toxicology 49 (2011) 991–998
rats (Tung et al., 2009). Kessova and Cederbaum (2003), have re-
ported that CYP2E1 is an effective generator of reactive oxygen
species such as the superoxide anion radical and hydrogen perox-
ide, and in the presence of iron catalysts, produces powerful oxi-
dants such as the hydroxyl radical. Lindane increases the level of
oxidative stress by diminution of antioxidant enzymes and the
present study reveals that the gallic acid could restore the antiox-
idant functions to near normal. The protective effect of gallic acid
may be attributed to its inhibiting activity on CYP2E1 thereby
reducing the level of bioactivation of lindane by the same CYP
isoform.
Moreover, there are reports in the literature regarding struc-
ture activity relationship for gallic acid viz., free radical scaveng-
ing efficiency of Gallic acid derivatives (GADs) is critically
dependent on the presence of the phenolic hydroxyl groups and
the steric freedom available in the molecule (Lu et al., 2006). As
GADs tend to deprotonate to give birth to GAD anions, which
has big influence on the radical-scavenging behaviors of GADs.
Gallic acid derivatives can efficiently scavenge free radicals, which
is partially responsible for their cytoprotective effects (Ji et al.,
2006). The decline in the levels of GSH, GST and GPx on lindane
administration may be due to the involvement of these enzymes
in the detoxification process of lindane in liver and kidney. And
the insufficient level of these antioxidants may damage the cells
via oxidative stress caused by lindane. By enhancing these antiox-
idants, and thereby decreasing the lipid peroxidation, gallic acid
protects the cells.
In conclusion, it may be mentioned that the altered biochemical
profiles due to lindane exposure is reversed towards normalization
by gallic acid. Gallic acid not only protects the integrity of plasma
membrane but at the same time increase the regenerative and
reparative capacity of the liver and kidney. Gallic acid may directly
combine with free radicals and lead to their inactivation which
may suppress the intracellular concentration of free radicals. These
results suggest that the compound gallic acid efficiently protects
the liver and kidney from oxidative damage by minimizing cell
membrane disturbances and helps in normal functioning of these
vital organs.
Conflict of Interest
The authors declare that there are no conflicts of interest.
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