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World Applied Sciences Journal 33 (6): 931-938, 2015 ISSN 1818-4952 © IDOSI Publications, 2015 DOI: 10.5829/idosi.wasj.2015.33.06.260

Effect of Red ( vulgaris L.) And its Fresh Juice Against Carbon Tetrachloride Induced Hepatotoxicity in Rats

Naeem M. Rabeh

Nutrition and Food science Dept., Faculty of Home Economics, Helwan University, Cairo, Egypt

Abstract: The present study aimed to investigate the effect of dried, fresh juice and waste (pulp) after juicing of red beetroot against carbon tetrachloride induced hepatotoxicity in rats. Total phenol, total flavonoids and antioxidant activity of the three mentioned samples were determined. The results from chemical analysis revealed that dried beet root have the highest amount of total phenols, total flavonoids and antioxidant activity followed by beet juice then beet waste respectively. Thirty male albino rats were divided into 5 groups;

group (1) control negative, while the other rats were administered a dose of (2 mL CcL4 /kg b.wt.) twice a week for two weeks to induce chronic damage in the liver then classified into 4 subgroups as follow, the 1st subgroup served as control positive, the 2nd subgroup was fed on basal diet supplemented with dried red beetroot at the level of 10%, the 3rd subgroups was given orally (10 ml/Kg b.wt./day) fresh juice from beetroot ( divided into 3 doses /day), the 4th subgroup was fed on basal diet and supplemented with the waste (pulp) from beetroot at the level of 10% for 8 weeks. The results from serum analysis indicated that supplementation with dried beetroot, juice and waste (pulp) of beetroot significantly (P<0.05) restored the enzyme activities of the liver AST, ALT and ALP to normal level. The mean values of MDA and serum total bilirubin were also significantly reduced (P< 0.05), while the mean value of total protein, albumin, CAT, SOD and GSH was significantly (P<0.05) increased as compared to control positive. Due to the presence of both total flavonoids and total phenols in beet root, juice and the waste of beetroot, these materials represent a rich sources of antioxidants and protect the liver cells from CcL4 induced liver damage. So, it is advice to add beet root powder and the waste (pulp) of beet root to bakery product, candies and yogurt and consume it as a routine diet to hepatic disease patients. Also, patients suffering from liver diseases may drink beet root juice to enhancing liver functions and increase antioxidant enzymes.

Key words: Hepatotoxicity Beetroot Carbon Tetrachloride Rats Antioxidants Liver Functions

INTRODUCTION food and as a source of natural dye [3]. Beets are small herbaceous with broad dark green . Chemical-induced liver injury depends mostly on Beet pigments, , have been examined as the oxidative stress in hepatic tissue and underlies the natural colorants in food products such as processed pathology of numerous diseases. There is still a lack of meat, ice cream, baked goods, candies and yogurt. effective therapeutics; hence, a treatment with Beetroot is a rich source of potent nutrients including antioxidants has been proposed for prevention and/or magnesium, , potassium, C and attenuation of injury. Such an approach has been defened betaine. Results from several in vitro studies have as chemoprevention and a large body of evidence from demonstrated that betalains from possess various experiments has supported its effecacy [1]. powerful antiradical and antioxidant activity [4]. Foods rich in antioxidants have been proposed as a tool Medicinally, the roots and leaves of the beet have been to prevent and cure liver damage [2]. employed as a folk remedy to treat a wide variety of Red beet (Beta vulgaris L.) is cultivated ailments including immune system stimulation and liver throughout the world for its roots, which are used as a and kidney diseases [5].

Corresponding Author: Naeem M. Rabeh, Nutrition and Food science Dept., Faculty of Home Economics, Helwan University, Cairo, Egypt. Tel: +201141048010. 931 World Appl. Sci. J., 33 (6): 931-938, 2015

Besides other active chemicals, beetroots contain a Rats: Thirty male rats of Sprague Dawley strain weighing unique class of water-soluble, nonphenolic antioxidants, (180 ±5 g.) were obtained from the National research the betalains, including two classes of compounds, Center, Dokki, Cairo. The rats were maintained under red betacyanins (principally ) and yellow standard laboratory conditions in an air conditioned betaxanthines. The antioxidant effects of betalains have room and housed in stainless steel cages one per cage been demonstrated mainly in various in vitro experiments at temperature 22±3 °C and relative humidity 30-70 %. [6]. Kapadia et al., [5] showed that co-administration of The animal diet was given ad libitum. Animals were betanin with model carcinogens exhibited a acclimatized for one week prior to experiment. chemopreventive effect on experimental carcinogenesis. The effect was evident as a significant reduction in tumor Preparation of Dried Beetroot, Juice and Waste of incidence, multiplicity and delay in tumor latency period. Beetroot: Red beetroots were washed with tap water, Agarwal et al., [7] reported the hepatoprotective chopped into small pieces and then dried at Solar Energy Center, National research Center, Dokki, Cairo. The dried activity of beetroot ethanolic extract against CcL4 induced liver injury in rats assessed on the basis of routine serum material was reduced into powder form as far as possible. markers of liver function, alanine aminotransferase The juice was prepared from chopped beetroots by (ALT), alkaline phosphatase (ALP) activities and the household juice extractor then filtered through stainless concentration of cholesterol and triglycerides. Also, steel refinery. The pulp left after juice extraction was dried Kujawska, et al., [8] investigated the potential protective in an oven at 50 ± 1°C and powdered using a lab grinder effect of beetroot juice (8 mL/kg/day for 28 days) in a and stored at 4°C till use. model of oxidative stress induced by N- nitrosodiethylamine and carbon tetrachloride and Determination of Antioxidant Content in Dried and Waste of Beetroot after Juicing: Total phenolic content mentioned that, pretreatment with beetroot juice can was determined according to the Folin-Ciocalteu counteract, to some extent, xenobiotic-induced oxidative procedure [10] and expressed as mg of gallic acid stress in rats. equivalent (GAE) per g of sample. Total flavonoid content Residue from the processing of fruits and , was determined according to Zilic et al. [10] and traditionally considered as an environmental problem, expressed as mg of catechin equivalent (CE) per g of are being increasingly recognized as sources for obtaining sample. Determination of radical DPPH scavenging high-phenolic products. The polyphenolics from waste activity (Antioxidant activity) was determined using the materials, being derived from agro-industrial production, stable 1,1-Diphenyl-2-picryl-hydrazyl (DPPH ) according may be used as functional food ingredients and as natural to Hwang and Do Thi [11]. The same methods were used antioxidants to replace their synthetic equivalents that for determination of antioxidant content in juice of have experienced growing rejection [9]. Conventional beetroot with some modifications (using the juice as an drugs and synthetic antioxidants used in the treatment of extract). liver diseases are generally inadequate and have some serious adverse effects. The interest for searching of Experimental Design: The basal standard diet was alternative drugs and natural antioxidants of prepared in accordance with AIN-93 formulation [12]. origin for the treatment of liver disease has increased. So, After a period of adaptation on basal diet, thirty rat were the aim of this work is to investigate the protective effect divided into five groups (6 rats each ); Group (1): was fed of dried, fresh juice and waste after juicing of red beetroot on basal diet only and served as a negative control group, (Beta vulgaris L.) against carbon tetrachloride induced while the other rats were administered a dose of (2 mL hepatotoxicity in rats. CcL4 /kg b.wt.) twice a week for two weeks to induce chronic damage in the liver according to Sundaresan and MATERIALS AND METHODS Subramanian, [13] then classified into four subgroups as follow, subgroup (1) served as control positive group, Chemicals: All chemicals including Carbon tetrachloride subgroup (2) was fed on basal diet supplemented with (CcL4 ) and Kits were purchased from local distributer dried red beetroot at 10%, subgroup (3) was given orally (Sigma chemical) Cairo, Egypt. (10 ml/Kg Body weight/day) fresh juice from beetroot ( divided to 3 doses /day), subgroup (4) was fed on basal Plant: Red beetroot (Beta vulgaris L.) was purchased diet supplemented with the waste from beetroot juice at from a local market, Cairo, Egypt. 10%.

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Biochemical Analysis: At the end of the experiment , betain and pigments in (eight weeks), the animals were fasted overnight, Beta vulgaris root of studies. The and then they were anaesthetized and sacrificed to antioxidant activity determined by the DPPH method obtain blood samples. Each blood sample was placed exhibited 40% and 78% activity in methanol extracts in dried clean centrifuge tube and then centrifuged of carrot and beetroot pulp waste (20 mg) respectively for 10 minutes at 3000 (rpm) to separate the serum. [19]. So, the results from chemical analysis suggest that Serum was carefully separated into clean dried red beetroot, juice and beetroot wastes can be exploited Wassermann tubes by using a Pasteur pipette and kept for their antioxidant components and used for value frozen until analysis. addition in food formulations. The effect of treatment with dried beetroot, juice from Statistical Analysis: The obtained results were red beetroot and residue from juice extraction (pulp) on statistically analyzed with SPSS Inc. software (version the activity of serum liver enzymes (ALT, AST, ALP and 16.0). One way ANOVA was used to study a significant total bilirubin) is recorded in Table (2). Subcutaneous difference between means of the dietary groups with a injection of CcL44 to rats in a dose (2 mL CcL /kg b.wt.) significance level at (P<0.05) [14]. twice a week for two weeks significantly (P < 0.05) elevated the mean levels of ALT, AST, ALP and total RESULTS AND DISCUSSION bilirubin when compared with the negative control group. In this aspect, Mansour et al. [20] reported that a single Table (1) shows the content of total phenols, total dose of CcL4 induced hepatotoxicity manifested flavenoids and the antioxidant capacity of dried, juice and biochemically by significant elevation of activities of liver waste (pulp) of red beetroot. The chemical analysis enzymes such as ALT and AST. Hepatic injury can be reveals that all three samples have total phenols and total determined by measuring the leakage of cellular enzymes flavonoids. Dried beet root have the highest amount of into plasma or serum. Increased serum enzyme levels, total phenols and total flavonoids followed by beet juice including ALT and AST had been associated with loss of then beet waste respectively. hepatocyte membrane integrity, or more specifically The DPPH is a decolorization assays which measure necrosis in the case of ALT elevation [21]. It is well the relative antioxidant abilities of natural extracts to established that CcL4 induces hepato-toxicity by scavenge free radicals generated in the assay system [15]. cytochrome P450 mediated reactions to produce Our results demonstrate that dried beetroot have the CcL4-derived radicals [22]. highest antioxidant activity followed by juice Beetroot On the other hand, bilirubin is a breakdown product then the waste after juicing. of hemoglobin which will conjugate with glucuronic The beet waste (pulp) has a considerable amount of acid in hepatocytes to increase its water solubility. total phenols, total flavonoids and the antioxidant Thus, release of unconjugated bilirubin from damaged or activity, so we have to use this waste in food technology. dead liver cells is also one of the major markers for liver The above results indicate that red beet roots have good damage [23]. antioxidant properties and were in accordance with Váli et The ALP is the prototype of these enzymes that al. [16] who mentioned that beetroots contains important reflects the pathological alteration in biliary flow. The bioactive agents (betaine and polyphenols), which have CcL4 induced elevation of this enzymatic acivity in the a wide range of physiologic effects. Pal et al., [17] and serum is in line with high level of serum bilirubins content Georgiev et al., [18] reported the presence of flavonoids, [24], this mechanism was in harmony with our results.

Table 1: Total phenols, total flavonoids and antioxidant activity content of dried, juice and waste (pulp) of red beetroot. Parameters ------Sample Total Phenols (GAE mg/g) Total Flavonoids (CE mg/g) DPPH (AAE mg/g) Dried beetroot 4.96 4.99 4.14 Beetroot juice 0.879 1.329 0.78 Beet waste (pulp) 0.233 0.287 0.105 GAE: Gallic acid equivalent, CE: Catchin equivalent, AAE: Ascorbic acid equivalent

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Table 2: Effect of dried, fresh juice and waste (pulp) from red beetroot on liver functions in carbon tetrachloride injected rats. Parameter ------Sample ALT ( /L) AST ( /L) ALP ( /L) Total Bilirbin (mg/dl) Control (-) 29.86 ± 2.12 c38.20 ± 1.42 c69.41 ± 2.44 bd4.60± 0.28 Control (+) 53.88 ± 1.78 a69.15 ± 2.28 a93.00 ±1.93 aa8.50 ± 0.25 Dried Beetroot 34.86 ± 7.79 bc 44.70 ± 1.57 b 69.16 ± 2.62 b 5.73 ± 0.24 c Beetroot Juice 36.50± 2.23 b47.66 ± 1.56 b71.00 ±1.86 bb7.19 ± 0.16 Beetroot waste (pulp) 38.91 ± 2.04 b49.16 ± 2.00 b73.50 ± 1.82 bb6.66 ± 0.25 *Values were expressed as Means ± SE, Values at the same column with different litters are significant at P<0.05.

Rats supplemented with either dried beetroot, juice Table 3: Effect of dried, fresh juice and waste (pulp) from red beetroot on and waste (pulp) from juice extraction had significantly serum total and albumin in carbon tetrachloride decreased (P<0.05) serum AST and ALT as compared injected rats. to positive control group with the most significant Parameters ------(P < 0.05) effect was observed in rats fed with dried Sample T. protein (g/dL) Albumin (g/dL) beet root. Also, these treatments induced suppression Control (-) 7.34 ± 0.20 aa3.61 ± 0.18 of the increased ALP activity (P < 0.05) with the Control (+) 5.79 ± 0.23 cd2.23 ± 0.18 concurrent depletion of raised bilirubins suggests the Dried Beetroot 7.01 ± 0.22 ab 3.30 ± 0.16 ab possibility of these materials to have ability to stabilize Beetroot Juice 6.68 ± 0.25 ab 2.88 ± 0.19 bc biliary dysfunction in rat liver during hepatotoxicity by Beet waste (pulp) 6.51 ± 0.18 b 2.68 ± 0.20 cd

CcL4 . *Values were expressed as Means ± SE, Values at the same column with Our results helped to stabilize the plasma membrane different litters are significant at P<0.05. and suppressed the leakage of enzymes through cellular membrane. This hepatoprotective effect may be Also, our results of the current study were in contributed by its antioxidant capacity [25]. accordance with those of Pal et al [17] who reported that, There were no significant differences in serum AST, the treatment with ethanolic extract of Beta vulgaris ALT and ALP among the three tested groups unless the caused significant reduction of CcL 4 induced elevated mean level of total bilirubin there was a significant (P serum levels of enzyme activities and bilirubin with < 0.05) difference between the rats fed dried beet and the parallel significant increase in total protein, indicating other both group fed on either beet juice or the residue of that, the extract could preserve the normal functional juice extraction (pulp). It was also clear that, there was no status of the liver. The hepato-protective effect of beet significant difference in the mean value of serum ALP root may be due to presence of its chemical contents. among the three tested group and the control negative The results were in agreement with Sadeek, [26] who group. reported that, treatment with beetroot (oral dose of Treatment with dried red beetroot, beet root juice and 8ml/Kg/day/ rat) for 28 days significantly (P<0.05) waste from juice extraction (pulp) significantly (P<0.05) restored the enzyme activities of the liver AST, ALT and restored the enzyme activities of the liver AST, ALT, ALP, ALP to normal level. The mean values of total bilirubin total bilirubin and total protein to normal level, these were also significantly reduced (P< 0.05). results were in accordance with results of Kapadia et al., From the results in Table (3) it could be observed [5] who recorded that red beetroot (Beta vulgaris L.) is that, administration of CcL4 caused a significant (P<0.05) used as a popular folk remedy for liver and kidney decrease in the serum levels of total protein and albumin diseases, for stimulation of the immune and hematopoietic as compared to the normal rats. Administration with dried systems and as a special diet in the treatment of cancer. beetroot, beet juice and waste from juice extraction (pulp) Also, Agarwal et al. [7] mentioned the hepatoprotective caused a significant (P<0.05) increase in serum level of activity of beetroot ethanolic extract against CcL4- total protein as compared to the positive control group. induced liver injury in rats assessed on the basis of Also the treatment with either dried beet root or beet juice routine serum markers of liver function, ALT and ALP significantly (P<0.05) increase the mean level of serum activities and the concentration of cholesterol and albumin, while beetroot waste (pulp) caused non triglycerides. significant differences as compared to control positive

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Table 4: Effect of dried, fresh juice and waste (pulp) from red beetroot on the activity of CAT, SOD, GSH enzymes and serum concentrations of MDA in carbon tetrachloride injected rats. Parameters ------Sample CAT (mmol/dl) SOD ( /dl) GSH ( mol/dl) MDA ( mol/dl) Cont (-) 68.25±1.75a99.75±1.79 a 43.17±0.13 ac1.82±0.13 Cont (+) 44.01±1.58d66.24±1.60 dd23.43±0.22 3.42±0.22 a Dried Beetroot 61.65±2.13b89.71±1.54 bb35.73±0.09 1.99±0.09 c Beet Juice 58.00±3.24b 85.30±1.88b 32.94±0.17bc 2.11±0.17c Beet waste (pulp) 50.50±1.55c72.37±0.94 cc28.41±0.12 2.61±0.12 b CAT: catalase, SOD: superoxide dismutase, GSH: reduced glutathione, MDA: malondialdehyde. *Values were expressed as Means ± SE, Values at the same column with different litters are significant at P<0.05. group. On the other hand, there was no significant activities. Also, these results were confirmed by Ruidong difference in the serum level of total protein among the et al., [34] and Wang et al., [35] who demonstrated that three tested group, while, treatment with dried beet root MDA levels in the CcL4 treated group as indicators of caused no significant difference in the mean level of lipid peroxidation were significantly higher than that in the serum albumin as compared to control negative group. normal control group. Due to dried beet root, juice and waste (pulp) from Glutathione (GSH) is an important endogenous beetroot enrich in phenolic antioxidants may provide a antioxidant substance. The decrease of GSH content may promising source of natural antioxidant, could protect be due to increased GSH consumption as it participates the liver against CcL4 induced liver toxicity. Phenolic in the detoxification system for the metabolism of compounds of plants fall into several categories; chief CcL4 and results in an enhanced susceptibility of among these are the flavonoids which have potent hepatocytes to CcL4 toxicity [36]. Our results showed that antioxidant activities [27]. Flavonoids have been shown CcL4 obviously decreased GSH content in the to be highly effective scavengers of most oxidizing hepatocytes, but treatment with either dried beetroot, molecules, including singlet oxygen and various free fresh juice and waste (pulp) from red beetroot radicals implicated in several diseases [28]. significantly (P<0.05) reverse it. This suggested that the Our results were in line with Bors et al., [29] who nature of supplementation with either dried beetroot, fresh mentioned that polyphenols and ?avonoid the large group juice and waste (pulp) from red beetroot protecting-SH of , are known to act as antioxidants. compounds (such as GSH) from CcL4 injury may be a Flavonoids are naturally occurring in plants and are mechanism of its hepatoprotection. thought to have positive effects on human health. Studies Supplemented diets with dried beetroot, fresh juice on flavonoidic derivatives have shown a wide range of and waste (pulp) from red beetroot caused significant antibacterial, antiviral, anti inflammatory, anticancer and increase (P<0.05) in the mean values of serum CAT, anti-allergic activities [30] and [31]. Also, Sadeek, [26] SOD and GSH level as compared with those of positive reported that, treatment with beetroot (oral dose of control groups. It is clear that, there were no significant 8ml/Kg/day/ rat) for 28 days significantly (P<0.05) differences in the mean values of serum CAT, SOD and increased the mean value of total protein and albumin. GSH level among groups fed on either dried beetroot and Results presented in table (4) demonstrate the effect beet juice group. But, there were a significant differences of dried beetroot, fresh juice and waste (pulp) from red (P<0.05) among groups fed on dried beetroot and group beetroot on the activity of catalase (CAT), superoxide fed on beet waste (pulp). dismutase (SOD), GSH enzymes and serum concentrations Liver cells possess a number of compensatory of malondialdehyde in carbon tetrachloride injected rats. mechanisms to deal with reactive oxygen species (ROS) The mean Values of serum CAT, SOD and GSH level in and their effects; among these are the induction of a untreated hepatotoxicity rats (control positive) group number of antioxidant such as SOD, CAT, were significantly lower at (P<0.05) as compared with glutathione peroxidase (GSHPx) and GSH. Therefore, those of the normal rats as a result of induction with CcL4 . oxidative stress, caused mainly by ROS, is also associated These results were in agreement with Augustyniak et al., with hepatic diseases [37]. SOD is an effective defense

[32] and Khan et al., [33] whom found that CcL4 initiates enzyme that protects biological macromolecules from lipid peroxidation and reduces tissue CAT and SOD oxidative stress [38].

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On the other hand, the elevations in the levels of end 3. Chevallier, A., 1996. The Encyclopedia of Medicinal products of lipid peroxidation (MDA) in the liver of rat Plants. New York: DK Publishing Inc., pp: 176. treated with CcL4 were significantly (P<0.05) observed as 4. Kujala, T.S., M.S. Vienola, K.D. Klika, J.M. Loponen compared with those of the negative control rats. MDA is and K. Pihlaja, 2002. and phenolic an indicator to evaluate the levels of lipid peroxidation compositions of four beetroot (Beta vulgaris) and inflammation during liver damage [39]. The increase . Eur. Food Res. Technol., 214: 505-510. in MDA levels in liver suggests enhanced lipid 5. Kapadia, G.J., M.A. Azuine, R. Sridhar, Y. Okuda, peroxidation leading to tissue damage and failure of A. Tsuruta; E. Ichiishi, T. Mukainake, M. Takasaki, T. antioxidant defense mechanisms to prevent formation of Konoshima, H. Nishino and H. Tokuda, 2003. excessive free radicals [24]. Chemoprevention of DMBA-induced UV-B Treatment with dried beetroot, fresh juice and waste promoted, NOR-1-induced TPA promoted skin (pulp) from red beetroot significantly (P<0.05) reversed carcinogenesis and DEN-induced phenobarbital these changes in serum levels of MDA compared with promoted liver tumors in mice by extract of beetroot. positive control group. It is obvious that, there were no Pharmacol Res., Feb; 47(2): 141-8. significant differences in the mean values of serum MDA 6. Kanner, J., S. Harel and R. Granit Betalains, 2001. level among groups fed on either dried beetroot and beet A new class of dietary cationized antioxidants. J. juice group and control negative group. These results Agric. Food Chem., 49: 5178-5185. were in agreement with Sadeek, [26] who reported that, 7. Agarwal, M., V.K. Srivastava, K.K. Saxena and A. treatment with beetroot (oral dose of 8ml/Kg/day/ rat) for Kumar, 2006. Hepatoprotective activity of Beta 28 days significantly (P<0.05) decrease the mean values of vulgaris against CcL4-induced hepatic injury in rats. MDA and significantly increase (P< 0.05) serum total Fitoterapia, 77(2): 91-93. protein, erythrocyte SOD as compared with the respective 8. Kujawska, M., E. Ignatowicz, M. Murias, mean values for control positive group. Also, The current M. Ewertowska, K. Miko ajczyk and results were confirmed with Lima et al., [40] who J. Jodynis-liebert, 2009. Protective Effect of Red mentioned that, the antioxidant mechanisms in biological Beetroot against Carbon Tetrachloride-and N- systems include direct quenching free radicals to Nitrosodiethylamine-Induced Oxidative Stress in terminate the radical chain reaction, chelating transition Rats, J. Agric. Food Chem., 57: 2570-2575. metals, acting as reducing agents, or stimulating the 9. Zhou, S., Z. Fang, Y. Lu, J. Chen, D. Liu and X. Ye, antioxidative enzyme activities. Other compounds with 2009. Phenolics and antioxidant properties of antioxidant activity include glutathione and flavonoids bayberry (Myrica rubra Sieb. et Zucc.) pomace. which protect cells against oxidative stress [41]. Food Chem., 112: 394-399. CONCLUSIONS 10. Zilic S., A. Serpen, G. Akýllýoglu, M. Jankovic and V. Gökmen, 2012. Distributions of phenolic Due to the presence of both total flavonoids and total compounds, yellow pigments and oxidative enzymes phenols, beet root, juice and the beetroot waste represent in wheat grains and their relation to antioxidant rich sources of antioxidants. So, it is advice to add beet capacity of bran and debranned flour. Journal of root powder and the waste (pulp) of beet root to bakery Cereal Science, 56: 652-658. product and consume it as a routine diet to hepatic 11. Hwang, E.S. and N. Do Thi, 2014. Effects of Extraction disease patients. Also, patients suffering from liver and Processing Methods on Antioxidant Compound diseases may drink beet root juice to enhancing liver Contents and Radical Scavenging Activities of Laver functions and increase antioxidant enzymes. (Porphyra tenera). Prev. Nutr. Food Sci., 19: 40-48. 12. Reeves, R.G., F.H. Nielsen and G.C. Fahey, 1993. REFERENCES "AIN-93 Purified Diets for Laboratory Rodents". J. Nutr., 123(1): 1939-1951. 1. Gonzales, C.A., 2006. Nutrition and cancer: the 13. Sundaresan, S. and P. Subramanian, 2003. S- current epidemiological evidence. Br. J. Nutr., allylcysteine inhibits circulatory lipid peroxidation 96: S42-S45. and promotes antioxidants in N-nitrosodiethylamine 2. Morisco, F., P. Vitaglione, D. Amoruso, B. Russo, induced carc-inogenesis. Pol. J. Pharmacol., 55: 37-42. V. Fogliano and N. Caporaso, 2008. Foods and liver 14. SPSS, 1986. Statistical package for social science, health. Mol. Aspects Med., 29: 144-150. version 16. SPSS Inc., II. U.S.A.

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