Bioactive Food as Dietary Interventions for Liver and ...

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Bioactive Foods and Supplements for Protection against Liver Diseases S.K. Shukla, V. Kumar International Centre for Genetic Engineering and Biotechnology, New Delhi, India

ABBREVIATIONS ALP Alkaline phosphatase ALT Alanine transaminase AST Aspartate transaminase CCl4 Carbon tetrachloride GSH Reduced glutathione HBV Hepatitis B virus HCC Hepatocellular carcinoma HCV Hepatitis C virus

1. INTRODUCTION Liver is the largest and most important metabolic organ of the human body, having an average weight of 2–3 pounds, located in the upper right-hand side of the abdomen. It is the center of various metabolic reactions, which occur in the body. Liver performs over 500 important functions like conversion of food component to essential blood components, storage of vitamins and minerals, production of many critical proteins and minerals, maintenance of hormonal balance and metabolism, and detoxification of toxic wastes of the body. It produces bile, which facilitates the process of digestion. Liver also plays a significant role in protein, lipid, and carbohydrate metabolism (Worman, 1999).

1.1 Liver Diseases Any disorder or condition which leads to malfunctioning of liver is termed as liver disease. Liver diseases are mainly categorized into two types: acute and chronic liver diseases. Acute disease occurs rapidly and usually exists for a very short duration. Chronic diseases are usually long term and exist for a long time, usually more than 6 months. In the clinical context, chronic liver disease is a disease process of the liver that involves a process of progressive destruction and regeneration of liver parenchyma leading to fibrosis and cirrhosis (Crawford, 2007). The main causes of chronic liver diseases are shown in Figure 36.1. Bioactive Food as Dietary Interventions for Liver and Gastrointestinal Disease http://dx.doi.org/10.1016/B978-0-12-397154-8.00019-1

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Unknown 17%

HBV 11%

Others 5%

HBV and alcohol 3%

HCV 26% Alcohol 24%

HCV and alcohol 14%

Figure 36.1 Primary causes of chronic liver diseases in human (based on a report of Center for Disease Control, USA).

1.2 Chronic Liver Disease Burden The burden of gastrointestinal disorder is on increase in both developing as well as developed world and chronic liver diseases following the same trend. About 350 million people are chronically infected with hepatitis B virus (HBV; Lok et al., 2001) and about 170 million people are infected worldwide with hepatitis C virus (HCV; Daniel, 2009). Despite viral infections, increasing rates of obesity and alcohol consumption worldwide predict that burden of chronic liver diseases related to alcohol as well as nonalcoholic steatohepatitis are set to increase. People having chronic liver diseases are more susceptible to develop serious disease symptoms of human immunodeficiency virus infection (World Health Organization, 2002). The aim of this chapter is to provide concise information about the different hepatoprotective ingredients of the food supplementary materials like spices, vegetables, fruits, drinks, and food grains.

2. HEPATOPROTECTIVE FOOD INGREDIENTS AND SUPPLEMENTS Several foods and supplements, which are used in routine diet of several Asiatic nations, possess hepatoprotective activity. This chapter provides a brief outline of hepatoprotective activity of different foods and supplements.

2.1 Hepatoprotective Spices 2.1.1 Turmeric Turmeric (Curcuma longa) is a widely used spice in India and other neighboring countries. It belongs to the ginger family. The main bioactive compound isolated from turmeric is a yellow color pigment known as curcumin. Curcumin exhibits a wide range of pharmacological activities including anti-inflammatory, antitumor, antioxidative, and free radical scavenging activity. In vitro as well as in vivo animal studies indicate the hepatoprotective

Bioactive Foods and Supplements for Protection against Liver Diseases

effect of curcumin in case of different hepatotoxic stresses caused due to carbon tetrachloride (CCl4), pentobarbital, galactosamine, 1-chloro-2,4-dinitrobenzene, and paracetamol. Although there are no clinical trials yet, these studies provide a strong base for the use of curcumin in different hepatic disorders (Luper, 1999). Curcumin also exhibits chemopreventive effect against hepatocarcinogenesis. Curcumin also shows antiproliferative, antiinvasive, and proapoptotic activity in different hepatocellular carcinoma (HCC)-derived cell lines (Bae et al., 2006). Based on these studies, it may be said that curcumin is a nontoxic and potent hepatoprotective natural compound. 2.1.2 Coriander Coriander (Coriandrum sativum) is one of the most important herbs used as a spice and flavoring agent. This is an annual herb and member of family Apiaceae. Coriander fruit is mostly present in all kinds of mixed spices used in India and other Asian countries. Hepatoprotective activity of essential oil of coriander has been studied in CCl4-induced hepatotoxicity in rat. It shows significant free radical scavenging activity and improved production of different antioxidative enzymes (Samojlik et al., 2010). 2.1.3 Garlic Garlic (Allium sativum) is a member of onion family Alliaceae. Mainly plant bulb is used as a spice. Ezaela et al. (2009) have shown that fresh garlic extract provides hepatoprotection against acetocarmine-induced liver injury. Garlic extract seems to provide hepatoprotection by lowering the alanine transaminase (ALT), alkaline phosphatase (ALP), and albumin level. In combination with silymarin, a well-known hepatoprotectant and clove oil, garlic extract shows hepatoprotective activity in N-nitrosodiethylamine with CCl4 and lead-induced hepatotoxicity, respectively, by reducing serum ALT, aspartate transaminase (AST), ALP, reduced glutathione (GSH), glutathione-S-transferase, and glutathione-S-reductase levels (Ajai et al., 2009). Pharmacokinetics of specific components of garlic extract has not been studied well in detail. Overall, garlic provides a good level of hepatoprotection in preclinical animal models. 2.1.4 Red chili Red chili is a member of genus Capsicum and family Solanaceae. Fruit is mainly used as a spice in different culinary preparations. The main active constituent of chili is capsaicin (8-methyl-N-vanilyl-6-nonenamide) along with other chemical constituents. Lee et al. (2004) have shown that capsaicin acts as proapototic factor for cancerous liver cells and suggested that capsaicin may be beneficial in therapeutic intervention of hepatoma. Capsaicin has shown chemotherapeutic and chemopreventive effect in different experimental models. It also exhibits antiproliferative and proapoptotic properties in HCC cell lines (Kim et al., 2005). Thus, these studies show the potential hepatoprotective effect of red chili.

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2.1.5 Black pepper Black pepper (Piper nigrum) is an annual vine that belongs to family Piperaceae. Its dry fruits are used as a spice worldwide. Black pepper possesses antioxidant, antipyretic, and anti-inflammatory property (Duke et al., 2003). Recently, Liu et al. (2010) have shown the anti-inflammatory and anticancer property of black pepper. Black pepper treatment resulted in reduced level of cyclooxygenase and NFkB with reduction in lipid peroxidation. Ganesh and Chandrasekhara (1987) reported that dietary feeding of black pepper leads to increase in bile secretions. Thus, black pepper consumption seems to confer hepatoprotection (Table 36.1).

2.2 Hepatoprotective Fruits 2.2.1 Grape Grape is a perennial vine, member of genus Vitis. Vitis vinifera (grape) fruit is used as food and for making the jam, jellies, and so on. It is widely used for the preparation of different Table 36.1 Hepatoprotective Spices Name Figure

Molecular mechanism

References

Turmeric (Curcuma longa)

Antioxidative, antiinflammatory, proapoptotic, antiproliferative

Bae et al. (2006), Luper (1999)

Coriander (Coriandrum sativum)

Free radical scavenger

Samojlik et al. (2010)

Garlic (Allium sativum)

Reduces serum ALT and AST levels, antioxidant

Ajai et al. (2009), Ezaela et al. (2009)

Chili (Capsicum annuum)

Proapoptotic and antiproliferative to HCC cell lines

Kim et al. (2005)

Black pepper (Piper nigrum)

Antioxidant and antiinflammatory

Duke et al. (2003)

Bioactive Foods and Supplements for Protection against Liver Diseases

kinds of wine. Resveratrol (main bioactive constituent of grape) is a potent antiinflammatory and antioxidant molecule. Resveratrol treatment to hepatoma-bearing mice have shown dramatic reduction in tumor volume and increased survival of experimental rat (Carbo et al., 1999). In vitro studies have shown that resveratrol inhibits proliferation and acts as proapoptotic agent when administered to different hepatoma cell lines (Ciolino et al., 1998). These studies represent the promising potential of grape fruit in hepatoprotection. 2.2.2 Custard apple Custard apple (Annona squamosa) tree is a small shrub of genus Annona and family Annonaceae. The fruit is popularly known as custard or sugar apple. There are several reports about the antioxidative, anti-inflammatory, and analgesic function of custard apple extracts and its active constituents (Chavan et al., 2010). Hepatoprotective effect of custard apple extract has been studied in isoniazid- and rifampicin-induced hepatotoxic model in Wistar rats. It has shown improved histopathology and reduced bilirubin level along with AST and ALT levels. The treatment group also showed less hepatocytic necrosis and inflammation in comparison to the control group (Saleem et al., 2008). 2.2.3 Apple Apple (Pyrus malus) is the most widely used fruit across the world. Apple fruit tree is perennial and member of family Rosaceae. Recently, hepatoprotective effects of apple polyphenols have been shown in CCl4-induced acute liver damage in mice. Apple polyphenol treatment prevents increase in ALT and AST level in CCl4-induced hepatotoxicity. Apple polyphenol-treated mice also show reduced malondialdehyde, increased level of superoxide dismutase, reduced level of GSH, and reduced lipid peroxidation (Yang et al., 2010). Miura et al. (2007) have also shown the antiproliferative and antiinvasive effect of apple polyphenols in rat ascites hepatoma cell line AH109A. Apple polyphenol treatment reduced the growth and metastasis of liver tumor and reduced lipid peroxide levels in rats transplanted with AH109A cells. These in vitro as well as in vivo findings provide a strong basis for the hepatoprotective effect of apple. 2.2.4 Indian gooseberry Indian gooseberry (Phyllanthus amarus) is a deciduous tree of family Phyllanthaceae. Fruit of P. amarus is edible and used in different parts of the world. It is known as amla in Hindi. Hydro-alcoholic extract of Phyllanthus fruit have shown improved liver function and reversal of profibrogenic events in CCl4-induced hepatotoxicity in male Wistar rats (Tasduq et al., 2005). Phyllanthus has also shown promising potential in treatment of patients with chronic HBV infection. It has shown greater extent of HBsAg clearance and HBeAg seroconversion. Phyllanthus along with interferon shows a greater potential in treatment of chronic HBV infection in comparison of interferon alone

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(Liu et al., 2001). These studies altogether indicate a strong hepatoprotective and antiviral activity of Phyllanthus. 2.2.5 Pomegranate Pomegranate (Punica granatum) is a deciduous tree of genus Punica and family Lythraceae. Pomegranate juice intake results in reduced hepatic oxidative stress in experimental mouse model (Faria et al., 2007). The crude pomegranate juice exhibited a strong antioxidative and antiproliferative property in comparison to the purified polyphenol components of pomegranate juice. Pomegranate peel extract also posses a strong hepatoprotective activity in hepatic fibrosis induced by bile duct ligation in rats. The peel extract treatment can reduce serum AST, ALT, and lactate dehydrogenase level significantly in these animals (Toklu et al., 2007). Thus, pomegranate extracts appear to have a strong hepatoprotective property. 2.2.6 Sea buckthorn Sea buckthorn (Hippophae rhamnoides) is a deciduous small tree of family Elaeagnaceae. Main consumable part is berry-like fruit of sea buckthorn. Zhao et al. (1987) have shown the hepatoprotective effect of sea buckthorns in CCl4-induced liver injury in rat model. Gao et al. (2003) have shown that fibrotic patients treated with sea buckthorn extract show improvement in liver functions and a significant decrease in the levels of inflammatory cytokines like tumor necrosis factor-alpha, interleukin-6, and reduced serum ALT and AST levels in the treated patients (Table 36.2).

2.3 Hepatoprotective Vegetables and Grains 2.3.1 Carrot Carrot (Daucus carota) is a biennial herb of family Apiaceae. Edible part of carrot is root. Carrot is a rich source of several carotenoids and vitamin A. It has been widely used as remedy of jaundice in Europe (Nadkarni, 1976). Bishayee et al. (1995) have shown hepatoprotective effect of carrot extract using CCl4-induced liver injury in Wistar rats. Administration of carrot extract to experimental rats resulted in decreased bilirubin and urea levels. It also reversed the increased levels of acid phosphatase and acid ribonuclease along with glucose-6-phosphatase and cytochrome P-450. 2.3.2 Ivy gourd Ivy gourd (Coccinia grandis), also known as parval/kundru, is a perennial herbaceous vine of family Cucurbitaceae. Its fruit is widely used as a vegetable in different parts of the world. Alcoholic extracts of C. grandis fruit possess the hepatoprotective property. When CCL4-induced hepatotoxicity rats are treated with the different doses of the alcoholic extract, they have shown lower value of serum ALT, AST, and ALP. It also reduced the bilirubin level comparable with the well-known hepatoprotective agent silymarin (Vadivu et al., 2008).

Bioactive Foods and Supplements for Protection against Liver Diseases

Table 36.2 Hepatoprotective Fruits Name Figure

Molecular mechanism

References

Grape (Vitis vinifera)

Anti-inflammatory, antioxidant, proapoptotic

Carbo et al. (1999), Ciolino et al. (1998)

Custard apple (Annona squamosa)

Anti-inflammatory, antioxidant, reduces serum ALT, AST, and ASP levels

Chavan et al. (2010), Saleem et al. (2008)

Apple (Pyrus malus)

Reduces lipid peroxidation, antiproliferative

Miura et al. (2007)

Indian gooseberry (Phyllanthus amarus)

Improves liver function, seroclearance of HBsAg

Liu et al. (2001), Tasduq et al. (2005)

Pomegranate (Punica granatum)

Aantiproliferative, reduces serum AST, ALT, and LDH levels

Toklu et al. (2007)

Sea buckthorn (Hippophae rhamnoides)

Anti-inflammatory, antioxidant

Gao et al. (2003)

2.3.3 Sweet corn Sweet corn (Zea mays var. saccharata), also known as Indian corn, sugar corn, or pole corn, is a variety of maize. Recently, Guo et al. (2009), using internal liver injury mice model, have shown that corn peptides possess hepatoprotective activity in vivo. Specifically, hepatoprotective activity of corn peptide was found significant at a dosage of 600 mg kg
1 body weight, and its hepatoprotective activity is very close to well-known hepatoprotective herb Silymarin (50 mg kg
1, body weight). Molecular mechanism underlying its hepatoprotective activity mainly includes its free radical scavenging activity, antioxidant property, activation of antioxidant enzymes, and reduces nitric oxide production. So the corn meals may provide beneficial effects in different kinds of chronic liver diseases, which cause liver injury. 2.3.4 Soy Soybean (Glycine max) is a member of family Leguminosae. Soy protein has high antioxidant property (Hu et al., 2004). Soy isoflavones and daidzein treatment to hepatoma cells like HepG2 and Huh-7 led to increased catalase activity (Kampkotter et al., 2008).

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Table 36.3 Hepatoprotective Vegetables and Grains Name Figure Molecular mechanism

References

Carrot (Daucus carota)

Antioxidant, reduces bilirubin and urea level

Bishayee et al. (1995)

Ivy gourd (Coccinia grandis)

Reduces serum AST and ALT levels

Vadivu et al. (2008)

Sweet corn (Zea mays var. saccharata)

Reduces nitric oxide production, antioxidant

Guo et al. (2009)

Soy (Glycine max)

Antioxidant, antiproliferative, antiinflammatory

Hu et al. (2004), Kampkotter et al. (2008)

Interestingly, in a cohort-based study in Japan, it has been found that soy consumption may reduce the risk of HCC (Sharp et al., 2005). Thus, soybean consumption has a beneficial effect of hepatoprotection in spite of rich dietary proteins (Table 36.3).

2.4 Hepatoprotective Drinks 2.4.1 Green tea All type of teas is derived from the leaves of Camellia sinensis, a member of family Theaceae. It is most widely used drink worldwide for over 5000 years. Green tea provides hepatoprotection against variety of hepatotoxic agents like naphthoquinone and alcohol. Catechins are very powerful antioxidants (Luper, 1999). Catechins have also been used for hepatitis B treatment since 1976 (Rauch, 1986) leading to clearance of HBeAg in chronic hepatitis B patients (Suzuki et al., 1986). In animal model of viral hepatitis, it has been found that green tea extract significantly reduced the ALT, AST, and ALP level. Green tea extract has shown therapeutic as well as preventive effects in diethyl nitrosamine-induced HCC in rat model (Luper, 1999). All these studies suggest strong hepatoprotective function of green tea. 2.4.2 Coffee Coffee (Coffea arabica) is widely used brewed drink worldwide, member of family Rubiaceae. Coffee powder is roasted seed of coffee beans. Phenolics and melanoidins are main bioactive constituents of coffee brew. Coffee consumption has shown beneficial

Bioactive Foods and Supplements for Protection against Liver Diseases

Table 36.4 Hepatoprotective Drinks Name Figure

Molecular mechanism

References

Tea (Camellia sinensis)

Antioxidant, reduces ALT, AST, and ALP levels

Luper (1999)

Coffee (Coffea arabica)

Antioxidant, inhibits HBV replication

Wang et al. (2009)

effect in different kinds of liver diseases like cirrhosis, HBV and HCV infections, and HCC. Coffee extract and its constituents have shown inhibitory effect on HBV replication in HepG2.2.25 cells as well as in duck HBV replication model (Wang et al., 2009). In another study, Freedman et al. (2009) have reported that coffee consumption results in slower disease progression in HCV-infected individuals as compared to non-coffee consumers. All these studies support the potent hepatoprotective function of coffee (Table 36.4).

3. CONCLUSIONS The herbal remedies and bioactive components of different plant products are taking center stage in the treatment of several liver diseases. Several phytochemicals present in food ingredients possess potential ability to prevent or reverse different kinds of liver injuries. Although underlying mechanism is not well understood, they represent a huge currency in terms of hepatoprotection. Although a large number of studies have been conducted at preclinical level, and many crude extract/molecule have been identified as potent hepatoprotective agent, more clinical studies are required for validation. It is high time to widen the horizon of study of hepatoprotective function of unexplored bioactive food and reap its benefit for mankind.

REFERENCES Ajai, G.O., Adeniyi, T.T., Babayemi, D.O., 2009. Hepatoprotective and some haematological aspects of Allium sativum and vitamin C in lead-exposed Wistar rats. International Journal of Medicine and Medical Sciences 1, 64–67. Bae, M.K., Kim, S.H., Jeong, J.W., et al., 2006. Curcumin inhibits hypoxia induced angiogenesis by down regulation of HIF-1. Oncology Report 15, 1557–1562. Bishayee, A., Sarkar, A., Chatterjee, M., 1995. Hepatoprotective activity of carrot (Daucus carota L.) against carbon tetrachloride intoxication in mouse liver. Journal of Ethnopharmacology 47, 69–74. Carbo, N., Costelli, P., Baccino, F.M., et al., 1999. Resveratrol, a natural product present in wine, decreases tumor growth in a rat tumor model. Biophysical Biochemical Research Communication 254, 739–743.

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Chavan, M.J., Wakte, P.S., Shinde, D.B., 2010. Analgesic and anti-inflammatory activities of 18-acetoxyent-kaur-16-ene from Annona squamosa L. Bark. Inflammopharmacology 18, 135–141. Ciolino, H.P., Daschner, P.J., Yeh, G.C., 1998. Resveratrol inhibits transcription of CYP1A1 in vitro by preventing activation of the aryl hydrocarbon receptor. Cancer Research 58, 5707–5712. Crawford, J.M., 2007. Basic mechanism in hepatopathology. In: Roderick, N.M., Burt, D.A., Portmann, B., Ferrell, L.D. (Eds.), Pathology of the Liver. fifth ed. Elsevier, Churchill Livingstone, pp. 119–146. Daniel, L., 2009. Global burden of hepatitis C. Liver International 29, 74–81. Duke, J.A., Bogensch-Godwin, M.J., deCellier, J., Duke, P.K., 2003. Piper nigrum L.(Piperaceae) black pepper. In: CRC Hand Book of Medicinal Spices. CRC Press, Washington, DC, pp. 253–263. Ezaela, C.C., Nweke, I.N., Unekwe, P.C., 2009. Fresh garlic extract protects the liver against acetaminophen-induced toxicity. The Internet Journal of Nutrition And Wellness 7 (1), 23–26. Faria, A., Monterio, R., Azevedo, I., Calhau, C., 2007. Effect of pomegranate (Punica granatum) juice intake on hepatic oxidative stress. European Journal of Nutrition 46, 271–278. Freedman, N.D., Everhart, J.E., Lindsay, K.L., et al., 2009. Coffee intake is associated with lower rates of liver disease progression in chronic hepatitis C. Hepatology 50, 1360–1369. Ganesh, B.B., Chandrasekhara, N., 1987. Effect of black pepper and piperine on bile secretion and composition in rats. Nahrung 31, 913–916. Gao, L., Gu, X.H., Cheng, F.T., Jiang, F.H., 2003. Effect of sea buckthorn on liver fibrosis: a clinical study. World Journal of Gastroenterology 9, 1615–1617. Guo, G., Sun, J., He, H., et al., 2009. Antihepatotoxic effect of corn peptides against Bacillus Calmette– Guerin/lipopolysaccharide-induced liver injury in mice. Food and Chemical Technology 47, 2431–2435. Hu, C.C., Hsiao, C.H., Huang, S.Y., et al., 2004. Antioxidant activity of fermented soybean extract. Journal of Agricultural and Food Chemistry 52, 5735–5739. Kampkotter, A., Wiegand, C., Timpel, C., et al., 2008. Increased expression of catalase in human hepatoma cells by the soy isoflavone, daidzein. Basic & Clinical Pharmacology & Toxicology 102, 437–442. Kim, J.A., Kang, Y.S., Lee, Y.S., 2005. A phospholipase-C dependent intracellular Ca2þ release pathway mediates the capsaicin-induced apoptosis in HepG2 human hepatoma cells. Archives of Pharmacal Research 28, 73–80. Lee, Y.S., Kang, Y.S., Lee, J.S., Nicolova, S., Kim, J.A., 2004. Involvement of NADPH oxidase mediated generation of reactive oxygen species in the apoptotic cell death by capsaicin in HepG2 human hepatoma cells. Free Radical Research 38, 405–412. Liu, J., Lin, H., McIntosh, H., 2001. Genus Phyllanthus for chronic hepatitis B virus infection: a systematic report. Journal of Viral Hepatology 8, 358–366. Liu, Y., Yadev, V.R., Aggarwal, B.B., Nair, M.G., 2010. Inhibitory effects of black pepper (Piper nigrum) extracts and compounds on human tumor cell proliferation, cyclooxygenase enzymes, lipid peroxidation and nuclear transcription factor-kappa-B. Natural Product Communications 5 (8), 1253–1257. Lok, A.S., Heathcote, E.J., Hoofnagle, J.H., 2001. Management of hepatitis B: 2000 – summary of a workshop. Gastroenterology 120, 1828–1853. Luper, S., 1999. A review of plants used in the treatment of liver disease: part two. Alternative Medicine Review 4, 178–188. Miura, D., Miura, Y., Yagasaki, K., 2007. Effect of apple polyphenol extract on hepatoma proliferation and invasion in culture and on tumor growth, metastasis, and abnormal lipoprotein profiles in hepatomabearing rats. Bioscience, Biotechnology, and Biochemistry 71, 2743–2750. Nadkarni, K.M., 1976. Indian Materia Medica, vol. 1. Popular Prokashan, Bombay. Rauch, G., 1986. The immune enhancing effect of cianidanol on macrophages and on the T-cell system. Methods and Findings in Experimental and Clinical Pharmacology 8, 147–150. Saleem, M.T.S., Christina, A.J.M., Chidambaranathan, N., et al., 2008. Hepatoprotective activity of Annona squamosa (Linn) on experimental animal model. International Journal of Applied Research in Natural Products 1, 1–7. Samojlik, I., Lakic, N., Mimica-Dukic, N., Dakovic-savicer, K., Bozin, B., 2010. Antioxidant and hepatoprotective potential of essential oils of coriander (Coriandrum sativum L.) and caraway (Carum carvi L.) (Apiaceae). Journal of Agricultural and Food Chemistry 15, 8848–8853.

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Sharp, G.B., Lagarde, F., Mizuno, T., et al., 2005. Relationship of hepatocellular carcinoma to soya food consumption: a cohort-based, case–control study in Japan. International Journal of Cancer 10, 290–295. Suzuki, H., Yamamoto, S., Hirayama, C., et al., 1986. Cinanidanol therapy for HBe antigen-positive chronic hepatitis: a multicentre, double blind study. Liver 6, 35–44. Tasduq, S.A., Mondhe, D.M., Gupta, D.K., Baleshwar, M., Johri, R.K., 2005. Reversal of fibrogenic events in liver by Emblica officinalis (Fruit), an Indian natural drug. Biological and Pharmaceutical Bulletin 28, 1304–1306. Toklu, H.Z., Dumlu, M.U., Sehirili, O., et al., 2007. Pomegranate peel extract prevents liver fibrosis in biliary obstructed rats. The Journal of Pharmacy and Pharmacology 59, 1287–1295. Vadivu, R., Krithika, A., Biplab, C., et al., 2008. Evaluation of hepatoprotective activity of the fruits Coccinia grandis Linn. Indian Journal of Health Research 1, 163–168. Wang, G., Shi, L., Ren, Y., et al., 2009. Anti-hepatitis B virus activity of chlorogenic acid, quinic acid and caffeic acid in vivo and in vitro. Antiviral Research 83, 186–190. World Health Organization, 2002. Diet, Physical Activity and Health. World Health Organization, Geneva. Worman, H.J., 1999. The Livers Disorders Source Book. Lowell house, Los Angles, USA. Yang, J., Li, Y., Wang, F., Wu, C., 2010. Hepatoprotective effect of apple polyphenols on CCL4 induced acute liver damage in mice. Journal of Agricultural and Food Chemistry 10, 6525–6531. Zhao, T.D., Cheng, Z.X., Liu, X.Y., et al., 1987. Protective effect of the sea buckthorn oil for liver injury induced by CCl4. Zhongcaoyao 18, 22–24.

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