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2014 30: 910 originally published online 16 November 2012Toxicol Ind HealthMihdiye Pirinççioglu, Göksel Kizil, Murat Kizil, Zeki Kanay and Aydin Ketani

induced oxidative stress in rats−The protective role of pomegranate juice against carbon tetrachloride  

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Article

The protective role of pomegranatejuice against carbon tetrachloride–induced oxidative stress in rats

Mihdiye Pirinccioglu1, Goksel Kızıl1, Murat Kızıl1,Zeki Kanay2 and Aydın Ketani3

AbstractMost pomegranate (Punica granatum Linn., Punicaceae) fruit parts are known to possess enormous antioxidantactivity. The present study was carried out to determine the phenolic and flavonoid contents of Derikpomegranate juice and determine its effect against carbon tetrachloride (CCl4)-induced toxicity in rats. Ani-mals were divided into four groups (n ¼ 6): group I: control, group II: CCl4 (1 ml/kg), group III:CCl4 þ pomegranate juice and group IV: CCl4 þ ursodeoxycholic acid (UDCA). Treatment duration was4 weeks, and the dose of CCl4 was administered once a week to groups II, III and IV during the experimentalperiod. CCl4-treated rats caused a significant increase in serum enzyme levels, such as aspartate amino-transferase, alanine aminotransferase and total bilirubin, and decrease in albumin, when compared with con-trol. Administration of CCl4 along with pomegranate juice or UDCA significantly reduces these changes.Analysis of lipid peroxide (LPO) levels by thiobarbutiric acid reaction showed a significant increase in liver,kidney and brain tissues of CCl4-treated rats. However, both pomegranate juice and UDCA prevented theincrease in LPO level. Histopathological reports also revealed that there is a regenerative activity in the liverand kidney cells. Derik pomegranate juice showed to be hepatoprotective against CCl4-induced hepatic injury.In conclusion, present study reveals a biological evidence that supports the use of pomegranate juice in thetreatment of chemical-induced hepatotoxicity.

KeywordsPomegranate juice, ursodeoxycholic acid, carbon tetrachloride toxicity, lipid peroxidation, antioxidant activity

Introduction

Free radicals play an important role in some pathogen-

esis of serious diseases, such as neurodegenerative

disorders, cancer, liver cirrhosis, cardiovascular

diseases, atherosclerosis, cataracts, diabetes and inflam-

mation (Aruoma, 1998). Many naturally occurring

antioxidative compounds from plant sources have been

identified as free radical inhibitors, active oxygen

scavengers or reducing agents (Duh, 1998; Yen and

Duh, 1994). Antioxidant compounds that can scavenge

free radicals have great potential in ameliorating these

diseases (Kirakosyan et al., 2003). However, synthetic

antioxidants (e.g., butylated hydroxyanisole and buty-

lated hydroxytoluene) can cause lung damage or pro-

mote the action of some carcinogens (Hocman, 1988).

Therefore, research on natural antioxidants has been

shown to play an important role in disease prevention.

The pomegranate (Punica granatum L.) is one of

the oldest edible fruits and is widely grown in many

tropical and subtropical countries (Salaheddin and

Kader, 1984). Recently, the antioxidant activity of

ethyl acetate, methanol and water extracts of pome-

granate peels and seeds has been reported in various

in vitro models (Singh et al., 2002). Pomegranate peel

1 Chemistry Department, University of Dicle, Diyarbakır, Turkey2 Physiology Department, Faculty of Veterinary Science,University of Dicle, Diyarbakır, Turkey3 Histology and Embryology Department, Faculty of VeterinaryScience, University of Dicle, Diyarbakır, Turkey

Corresponding author:Murat Kızıl, Department of Chemistry, Faculty of Science,University of Dicle, Diyarbakır 21280, Turkey.Email: muratk@dicle.edu.tr

Toxicology and Industrial Health2014, Vol. 30(10) 910–918© The Author(s) 2012Reprints and permissions:sagepub.co.uk/journalsPermissions.navDOI: 10.1177/0748233712464809tih.sagepub.com

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extracts have also been shown to possess in vivo antiox-

idant activity by Kotamballi et al. (2002). Pomegranate

juice significantly reduced atherosclerotic lesion areas

in immune-deficient mice and intima–media thickness

in cardiac patients during medications. It also decreased

lipid peroxidation in patients with type 2 diabetes and

systolic blood pressure and serum angiotensin-

converting enzyme activity in hypertensive patients

(Basu and Penugonda, 2009). Celik et al. (2009) have

reported the hepatoprotective role and antioxidant

capacity of pomegranate flowers infusion against tri-

chloroacetic acid (TCA)-exposed rats. They have found

that administration of subacute TCA promotes malon-

dialdehyde (MDA) concentration fluctuates in the anti-

oxidative systems and elevates tissue damage serum

marker enzymes the plant beverage supplement impart

protection against carcinogenic chemical-induced liver

injury and oxidative stress (Celik et al., 2009). There-

fore, in the last few years, the consumption of this fruit

and its products has increased, the juice being the most

popular form (Viuda-Martos et al., 2010).

Liver is an important organ actively involved in

metabolic functions and is a frequent target of a num-

ber of toxicants. One of the major functions of the

liver is the detoxification of xenobiotics and toxin

(Mitra et al., 1998). Carbon tetrachloride (CCl4) is

one of the most commonly used hepatotoxins in the

experimental study of liver diseases (Priya et al.,

2011). CCl4 intoxication in various studies has

demonstrated that CCl4 causes free radical generation

in many tissues such as liver, kidney, hearth, lung,

testis, brain and blood (Cemek et al., 2010; Dashti

et al., 1989). Colchiceine and ursodeoxycholic acid

(UDCA) are drugs currently in use as therapy for

different types of liver damage. Previous studies

investigated their ability to reverse the damage

induced by CCl4 in rats (Altas et. al., 2011; Nava-

Ocampo et al., 1997).

Pomegranate is widely consumed as fresh fruit and

juice. The use of pomegranate fruit dates from ancient

times, and reports of its therapeutic qualities have

echoed throughout the ages. Turkey is one of the main

pomegranate-producing countries in the world. It is

possible to grow pomegranates in most parts of

Turkey. Although Turkey’s total pomegranate pro-

duction changes from year to year, recent production

has reached 100,000 tons/year (Ercisli et al., 2007).

The antioxidant capacity of eight pomegranate juices

(Izmir 8, Izmir 10, Izmir 23, Izmir 26, Izmir 1264,

Izmir 1479, Izmir 1499 and Zivzik) was studied by

Cam et al. (2009). Six pomegranate (Dikenli ince

kabuk, Eksi, Kan, Katirbasi, Serife and Tatli) culti-

vars obtained from various sites from the Mediterra-

nean Region of Turkey were also evaluated for

antioxidant properties by Ozgen et al. (2008). How-

ever, there are no studies on Derik pomegranate that

is one of the pomegranate cultivars grown in South

East Turkey. Derik pomegranate fruit is round with

leathery rind and exhibits yellow to deep pink color.

There has been a virtual explosion of interest in the

pomegranate as a medicinal and nutritional product

because of its multifunctionality and its great benefit

in the human diet because it contains several groups

of substances that are useful in disease risk reduction.

As a result, the field of pomegranate research has

experienced tremendous growth (Martinez et al.,

2006). The present study was conducted to determine

the phenolic and flavonoid contents of Derik pome-

granate juice and to determine its effect against

CCl4-induced toxicity in rats. The present study was

also planned to compare the protective effect of

pomegranate juice and UDCA against liver, kidney

and brain damage induced by CCl4 in rats.

Materials and methods

Materials and chemicals

This study was approved by Scientific and Ethics

Committee of the Medical Science Application and

Research Center of Dicle University (DUSAM,

Diyarbakir, Turkey). The rats were obtained from

DUSAM. Pomegranate was harvested in Derik dis-

trict, Mardin, Turkey. CCl4, TCA, 2-thiobarbituric

acid (TBA), gallic acid, quercetin and 1,1,3,3-

tetramethoxypropanol were purchased from Sigma-

Aldrich (St. Louis, Missouri, USA), Folin-Ciocalteu’s

phenol reagent was obtained from Merck (Darmstadt,

Germany). UDCA was obtained from Dr. Falk

Pharma GmbH (Freiburg, Germany).

Preparation of pomegranate juice

Fresh pomegranate fruits (P. granatum L., variety of

Derik) were obtained from Derik, Mardin, Turkey,

on October 2010. They were washed, drained and cut

into halves. The ruby seeds and all white pulpy part were

together squeezed with electrical blender. The juice was

then stored in 1 ml aliquots at �20�C until use.

Total phenolics contents

The amount of total polyphenols in the fresh pome-

granate juice was determined according to the

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Folin-Ciocalteau method (Slinkard and Singleton,

1977). Gallic acid equivalent (GAE) was used as a

calibration standard, and results were expressed as

GAEs (microgram GAE per milliliter of juice

sample). The absorbance was measured using the

ultraviolet–vis spectrophotometer at the wavelength

of 765 nm. The concentration of phenolic compounds

was calculated by the following equation that was

obtained using standard gallic acid curve:

Absorbance ðAÞ ¼ 0:0025� gallic acid ðmgÞ;R2 ¼ 0:9980:

Total flavonoid contents

Total flavonoid content of pomegranate juice was

determined by a colorimetric method based on the

formation of flavonoid–aluminum complex, using

quercetin as a standard (Djeridane et al., 2006).

Briefly, 1 ml of different concentrations of quercetin

solution (15–75 mg/ml) and pomegranate juice was

added to different test tubes containing 0.1 ml of

10% aluminum nitrate, 0.1 ml of 1 M potassium

acetate and 3.8 ml of methanol. The mixture were

thoroughly mixed and incubated in a water bath for

40 min at 25�C. The absorbance of the mixtures was

measured at 415 nm using a spectrophotometer. Three

replicates were made for each test sample. The total

flavonoid content was expressed as microgram

quercetin equivalents (QEs) per milliliter watermelon

juice. The concentration of flavonoid compounds was

calculated by the following equation that was

obtained using standard quercetin curve:

Absorbance ðAÞ ¼ 0:0132� quercetin ðmgÞ;R2 ¼ 0:9998:

Animals and treatments

Male Wistar rats (150–200 g) were obtained from

Central Animal House, Dicle University. The animals

were grouped and housed in polyacrylic cages with

not more than six animals per cage and maintained

under standard laboratory conditions with dark and

light cycle. They were allowed free access to standard

pellet diet and water ad libitum. All procedures were

reviewed and approved by Institutional Animal

Ethical Committee. Rats were divided into four

groups of six animals each. The groups were named

as control group (group I), CCl4 group (group II),

pomegranate juice þ CCl4 group (group III) and

reference drug (UDCA) þ CCl4 group (group IV).

The animals of group I received a normal vehicle only

and groups II, III and IV were treated with CCl4 (1:1

in olive oil) at 1 ml/kg of body weight four times dur-

ing experimental period. Groups III and IV were also

given pomegranate juice (2 ml/kg of body weight) and

UDCA (10 mg/kg of body weight; Maton et al., 1977)

every day for 4 weeks, respectively. At the end of

4 weeks of experimental period, all animals were

killed by decapitation, blood samples were collected,

and their livers, brains and kidneys were removed

immediately and stored at 80�C until further analysis.

Biochemical analysis

Blood samples were centrifuged at 2000g for 10 min,

and serum was stored at 4�C for biochemical analysis.

The activities of serum aspartate aminotransferase

(AST), alanine aminotransferase (ALT), albumin

(ALB) and total bilirubin (TB) were assayed spec-

trophotometrically with an autoanalyzer (Unicee

DXC800, Synchron Clinical System, Beckman

Coulter, Ireland) according to the standard proce-

dures and using commercially available diagnostic

kits (Beckman Coulter, Galway, Ireland).

Determination of thiobarbituric acid reactivesubstances (TBARS) in the tissue samples

The liver, kidney and brain tissue samples were

homogenized with 120 mM KCl, 50 mM phosphate

buffer pH 7.4 (1:10, w/v). The homogenates were cen-

trifuged at 700g at 4�C for 10 min and the supernatant

was kept at �20�C until use. MDA level of liver, kid-

ney and brain tissue samples was measured by the

thiobarbituric acid reaction method (TBARS).

TBARS were determined calorimetrically (Draper

and Hadley, 1990). Briefly, 1 ml of each sample

was mixed with 1 ml of TCA 10% and 1 ml of

TBA 0.67% and then heated in a boiling water bath

for 15 min. Tubes were chilled on ice, and the

rose-colored trimethin complex was extracted into

3 ml of n-butanol. The organic phase was separated

by centrifugation for 10 min at 3000g; MDA, an

intermediate product of lipoperoxidation, was

determined by absorbance at 535 nm. A standard

curve for TBARS was prepared with 1,1,3,3-

tetramethoxypropanol in a concentration range of

0.1–10 nmol. The results are expressed as the

nanomole MDA per gram tissue.

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Histological procedure

The liver, kidney and brain samples were rapidly dis-

sected out, and tissue sections (5 mm) of liver, kid-

ney and brain were fixed by immersion at room

temperature in 10% neutral formalin solution. For

the histological examinations, paraffin-embedded

tissue sections of liver and kidney were stained

with trichrome masson (Tripple), and the brain tis-

sue samples were stained with hematoxylin–eosin.

The tissue samples were then examined and photo-

graphed under a light microscope (Nikon-Eclipse

400) for the observation of structural abnormality.

Statistical analysis

The parameter values were all expressed as the

mean + SD. Significant differences among the

groups were determined by one-way analysis of var-

iance using SPSS 12.0 software package program.

The results were considered significant if the value

of P was <0.05.

Results

Total phenolic and flavonoid

Table 1 shows the total phenolic and flavonoid con-

tents of Derik pomegranate juice. In the present study,

the amount of total phenolic and flavonoid were found

to be 3501.0 + 282.40 mg GAE/ml juice and

161.5 + 1.83 mg QUE/ml juice in Derik pomegranate

juice, respectively.

Serum marker enzymes

Table 2 presents the activities of serum AST, ALT,

TB and ALB in control and experimental rats. AST,

ALT, TB and ALB levels were found to be

62.66 + 6.50 IU/l, 143.60 + 23.71 IU/l,

0.25 + 0.09 mg/dl and 1.80 + 0.17 mg/dl in control

rats (group I), respectively. The results show that

CCl4 administration significantly increased the activ-

ities of AST and ALT levels in the serum of group II

when compared with groups I, III and IV. TB levels

were also increased from 0.25 + 0.09 mg/dl to

1.04 + 0.20 mg/dl in group II rats. This increase was

significantly higher than values obtained from the

other groups (groups I, III and IV). The level of ALB

was also significantly reduced in CCl4-treated rats

(group II) when compared with group I, III and IV

rats. ALB levels decreased from 1.80 + 0.17 mg/dl

to 1.03 + 0.20 mg/dl in group II rats. The group I

(control animals), group III (received pomegranate

juice) and group IV (received UDCA) did not show

any significant change in TB and ALB levels. The

effect of pomegranate juice was comparable with that

of UDCA in all the tested marker enzymes.

Lipid peroxidation

Lipid peroxidation was estimated in terms of TBARS,

using MDA as standard. Figure 1 shows that the level

of peroxidation products (MDA) in tissues of control

and experimental rats. In group I, MDA levels were

found to be 2.92 + 0.36, 4.56 + 0.32 and

4.80 + 1.11 nmol/g tissue in liver, kidney and brain

Table 1. Total phenolics and flavonoids content of Derik pomegranate juice.

Sample Total phenolic content (mg GAE/ml juice) Total flavonoid content (mg QUE/ml juice)

Derik pomegranate juice 3,501.0 + 282.40 161.5 + 1.83

QUE: quercetin equivalent; GAE: gallic acid equivalent.

Table 2. Hepatic markers in the serum of control and experimental rats.a

Groups ALT (IU/l) AST (IU/l) Bilirubin (mg/dl) Albumin (mg/dl)

Control 62.66 + 6.50a 143.60 + 23.71a 0.25 + 0.09a 1.80 + 0.17a

CCl4 (1 ml/kg) 3,242.66 + 289.31b 2,610.70 + 151.05b 1.04 + 0.20b 1.03 + 0.20b

Derik pomegranate juice (2 ml/kg) 2,410.33 + 144.50c 1,905.00 + 164.55c 0.52 + 0.05a 1.51 + 0.23a

UDCA (10 mg/kg) 2,238.67 + 108.74c 1,835.00 + 126.19c 0.44 + 0.15a 1.70 + 0.08a

UDCA: ursodeoxycholic acid; CCl4: carbon tetrachloride; AST: aspartate aminotransferase; ALT: alanine aminotransferase.aValues are given as mean + SD for six rats in each group. Means with different letters differ significantly, p < 0.05, whereas the valuessharing common letters are not significantly different, at p < 0.05.

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samples, respectively. A significant elevation in the

levels of MDA in liver (5.70 + 0.60 nmol/g tissue),

kidney (4.23 + 4.58 nmol/g tissue) and brain (7.60

+ 1.84 nmol/g tissue) samples were observed in

group II when compared with control group. The

treatment of Derik pomegranate juice resulted in a

significant decrease in MDA level in liver (3.01 +0.33 nmol/g tissue), kidney (8.67 + 2.63 nmol/g tis-

sue) and brain (5.02 + 1.09 nmol/g tissue) tissues

when compared with CCl4-treated rats. The liver

(2.80 + 0.34 nmol/g tissue), kidney (8.56 + 1.57

nmol/g tissue) and brain (5.23 + 0.77 nmol/g tissue)

tissues of UDCA-treated animals also showed a sig-

nificant decrease compared with CCl4-treated rats.

No significant difference was observed between

Derik pomegranate juice and UDCA in MDA levels

of liver, kidney and brain tissue samples.

Histopathology

In this study, the control rats (group I) showed normal

appearance of liver (Figure 2a). The liver samples of

CCl4-administered rats (group II) showed the focal

hepatocytes damage and degeneration (Figure 2b).

Vacuolization, fatty changes and necrosis of hepato-

cytes were severe in the centrilobular region. Derik

pomegranate juice-treated (group III; Figure 2c) and

UDCA (group IV; Figure 2d)-treated rats showed near

normal appearance hepatocytes with a mild degree of

fatty change, necrosis almost comparable to the con-

trol. The kidney samples of group I rats showed nor-

mal appearance of kidney (Figure 3a). The kidney

samples of group II rats showed the dilated tubules

with cloudy swelling (Figure 3b). Derik pomegranate

juice- (Figure 3c) and UDCA (Figure 3d)-treated rats

showed almost normal appearance with mildly dilated

tubules with regenerating epithelium in kidney. The

brain samples of all groups showed normal appear-

ance of brain structure (Figure 4(a) to (d)).

Discussion and conclusions

During recent years, there has been considerable

interest in identifying natural sources with antioxidant

activities to prevent oxidative stress-induced dam-

ages. Epidemiological findings have shown that con-

suming foods and beverages having high levels of

phenolic compounds decreases the risk of many dis-

eases such as cardiovascular and also protects against

certain forms of cancer (Arts and Hollman, 2005;

Graf et al., 2005).

liver kidney brain0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

Group 1Group 2Group 3Group 4

a

b

a a

a

b

c c

a

b

a a

nmol

MD

A/g

tiss

ue

Figure 1. MDA levels of studied organs in control and experimental rats. Data are expressed as mean + SD for an aver-age of six animals. Means with different letters differ significantly, p < 0.05, whereas the values sharing common letters arenot significantly different, p < 0.05. Group I: control, group II: CCl4, group III: CCl4 þ pomegranate juice and group IV:CCl4 þ ursodeoxycholic acid. CCl4: carbon tetrachloride; MDA: malondialdehyde.

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Pomegranate fruit has valuable compounds in dif-

ferent parts of the fruit whose functional and medic-

inal effects such as antioxidant, anticancer and

antiatherosclerotic effects have been confirmed

(Mertens-Talcott et al., 2006; Perze-Vicente et al.,

2002). In previous study, pomegranate peel methanol

extract was reported to possess high antioxidant activ-

ity in various models (Singh et al., 2002). Pomegra-

nate peel extracts also provide protection against

CCl4 toxicity (Kotamballi et al., 2002). CCl4 is one

of the most commonly used hepatotoxins in the experi-

mental study of liver diseases. Its metabolites such as

trichloromethyl radical (CCl3�) and trichloromethyl

peroxy radical (CCl3O2�) are involved in the pathogen-

esis of liver and kidney damage (Singh et al., 1999).

In this study, we aimed to investigate the inhibitory

effects of Derik pomegranate juice on CCl4-induced

lipid peroxidation and its effect on plasma ALT, AST,

ALB and bilirubin. Administration of CCl4 signifi-

cantly raised the serum level of the enzymes, such

as ALT, AST and bilirubin in rats. Pomegranate juice

and UDCA caused a decrease in the activity of the

ALT and AST. The level of ALB was also signifi-

cantly reduced in CCl4-treated rats when compared

with group I, II and III rats. The elevated activities

of AST, ALT, TB and ALB are indicative of cellular

Figure 2. Histological sections of livers. (a) Group I: normal rats, (b) group II: CCl4-treated rats, (c) group III: pomegra-nate juice þ CCl4-treated rats and (d) group IV: UDCA þ CCl4-treated rats. (V: vena centralis; H: hepatosit; stain: tri-chrome mason-Tripple.) Original magnifications are indicated in the figures. CCl4: carbon tetrachloride; UDCA:ursodeoxycholic acid.

Figure 3. Histological sections of kidneys. (a) Group I: normal rats, (b) group II: CCl4-treated rats, (c) group III: pome-granate juice þ CCl4-treated rats and (d) group IV: UDCA þ CCl4-treated rats. (G: glomerulus; C: cloudy swelling; stain:trichrome mason-Tripple.) Original magnifications are indicated in the figures. CCl4: carbon tetrachloride; UDCA: urso-deoxycholic acid.

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leakage and loss of functional integrity of cell mem-

branes in the liver (Goldberg and Watts, 1965). Our

results indicated that Derik pomegranate juice signif-

icantly prevented these changes and indicated

improvement in the functional status of the liver. In

the current study, MDA levels were found to be

significantly higher in CCl4-induced group of liver,

kidney and brain samples when compared with other

group of liver, kidney and brain samples. UDCA, a

hydrophilic bile acid with low-intrinsic toxicity, has

been successfully used in the treatment of cholestatic

liver diseases. Its therapeutic effects have been attrib-

uted to several mechanisms, including the ability to

stimulate hepatobiliary secretion and inhibit liver cell

apoptosis (Mas et al., 2008; Paumgartner and Beuers,

2002). Derik pomegranate juice and UDCA treatment

decreases MDA levels in all the tested tissues, indicat-

ing that the pomegranate juice has a protective action

against CCl4-induced toxicity. These findings were

also supported by the histopathological analysis of

liver and kidney tissues of groups 3 and 4. However,

the brain samples of all the groups showed normal

appearance of brain. CCl4 appears to cross the

blood–brain barrier; however, the toxicity of CCl4in brain is relatively lower than that in the kidney and

liver. This is probably due to limited dose of CCl4.

Pomegranate exhibits good antioxidant capacity

and is an effective scavenger for several reactive

oxygen species, primarily due to its high levels of

phenolic acids, flavonoids and other polyphenolic

compounds (Aviram et al., 2000). Our study also

showed that selected Derik cultivars have high

amount of total phenolics and flavonoids. Total

phenolic content was found to be higher than total fla-

vonoid content in Derik pomegranate juice. Phenolic

antioxidants are products of secondary metabolism in

plants and are good source of natural antioxidants in

human diets. The Derik pomegranate juice seems to

be a rich source of fruit containing large amount of

phenolic acids, so it is considered to be a promising

source of natural antioxidants. Administration of CCl4along with pomegranate juice or UDCA significantly

reverses biochemical changes in serum. Analysis of

lipid peroxide (LPO) levels by thiobarbutiric acid

reaction showed a significant increase in liver, kidney

and brain tissues of CCl4-treated rats. The hepatotoxic

effects of CCl4 are largely due to its active metabo-

lites CCl3� and CCl3O2

� (Srivastava et al., 1990). Both

the radicals can bind covalently to the macromole-

cules and induce peroxidative degradation of the

membrane lipids in endoplasmic reticulum rich in

polyunsaturated fatty acids (Recnagel, 1967). Free

radical-induced lipid peroxidation is believed to be

one of the major causes of cell membrane damage,

leading to a number of pathological situations (Slater,

1984). This leads to the formation of LPOs followed

by various changes in biochemical parameters. These

properties of pomegranate juice are thought to pro-

vide many beneficial effects against organ damages.

The health benefits of pomegranate have been attrib-

uted to its wide range of phytochemicals, which are

predominantly polyphenols, including primarily

hydrolyzable ellagitannins, anthocyanins and other

polyphenols. Both Derik pomegranate juice and

UDCA prevented the increase in the LPO level, which

was almost brought to normal range. It was concluded

Figure 4. Histological sections of brains. (a) Group I: normal rats, (b) group II: CCl4-treated rats, (c) group III: pomegra-nate juice þ CCl4-treated rats and (d) group IV: UDCA þ CCl4-treated rats (stain: hematoxylin–eosin). Original magni-fications are indicated in the figures. CCl4: carbon tetrachloride; UDCA: ursodeoxycholic acid.

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that Derik pomegranate juice may be used in CCl4-

induced toxicity to prevent lipid peroxidation and also

to prevent tissue damage. The consumption of pome-

granate has grown tremendously due to its reported

health benefits. Pomegranate and its derivates, such

as juice, peel, and seeds, are rich source of several

high-value compounds with potential beneficial phy-

siological activities. The rich bioactive profile of

pomegranate makes it a highly nutritious and a desir-

able fruit crop. The results of the present work indi-

cate the presence of compounds possessing high

antioxidant activity in Derik pomegranate juice. Fur-

ther studies are needed, however, with individual phe-

nolic compounds of Derik pomegranate juice to

elucidate the different antioxidant mechanisms and

possible synergism.

Funding

This work was supported by grants from Dicle University

Research Foundation (Project no.: 09-FF-44), Turkey.

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