Effect of dietary supplementation of grape skin and seeds on liver fibrosis induced by dimethylnitrosamine in rats

  • Shin, Mi-Ok (Department of Food and Nutrition, Silla University) ;
  • Moon, Jeon-Ok (College of Pharmacy, Pusan National University)
  • Received : 2010.02.03
  • Accepted : 2010.10.01
  • Published : 2010.10.31


Grape is one of the most popular and widely cultivated fruits in the world. Although grape skin and seeds are waste product of the winery and grape juice industry, these wastes contain large amounts of phytochemicals such as flavonoids, phenolic acids, and anthocyanidins, which play an important role as chemopreventive and anticancer agents. We evaluated efficacies of grape skin and seeds on hepatic injury induced by dimethylnitrosamine (DMN) in rats. Treatment with DMN significantly increased levels of serum alanine transaminase, aspartate transaminase, alkaline phosphatase, and bilirubin. Diet supplementation with grape skin or seeds (10% daily for 4 weeks) prevented these elevations. The grape skin and seeds also restored serum albumin and total protein levels, and reduced the hepatic level of hydroxyproline and malondialdehyde. Furthermore, grape skin and seeds reduced DMN-induced collagen accumulation, as estimated by histological analysis of liver tissue stained with Sirius red. Grape skin and seeds also reduced hepatic stellate cell activation, as assessed by ${\alpha}$-smooth muscle actin staining. In conclusion, grape skin and seeds exhibited in vivo hepatoprotective and antifibrogenic effects against DMN-induced liver injury, suggesting that grape skin and seeds may be useful in preventing the development of hepatic fibrosis.


Supported by : Pusan National University


  1. Friedman SL. Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver. Physiol Rev 2008;88:125-72.
  2. Moreira RK. Hepatic stellate cells and liver fibrosis. Arch Pathol Lab Med 2007;131:1728-34.
  3. Li D, Friedman SL. Liver fibrogenesis and the role of hepatic stellate cells: new insights and prospects for therapy. J Gastroenterol Hepatol 1999;14:618-33.
  4. Nassiri-Asl M, Hosseinzadeh H. Review of the pharmacological effects of Vitis vinifera (Grape) and its bioactive compounds. Phytother Res 2009;23:1197-204.
  5. Haggerty HG, Holsapple MP. Role of metabolism in dimethylnitrosamine- induced immunosuppression: a review. Toxicology 1990;63:1-23.
  6. Terracini B, Magee PN, Barnes JM. Hepatic pathology in rats on low dietary levels of dimethylnitrosamine. Br J Cancer 1967; 21:559-65.
  7. Weng HL, Cai WM, Liu RH. Animal experiment and clinical study of effect of gamma-interferon on hepatic fibrosis. World J Gastroenterol 2001;7:42-8.
  8. Lee ES, Lee HE, Shin JY, Yoon S, Moon JO. The flavonoid quercetin inhibits dimethylnitrosamine-induced liver damage in rats. J Pharm Pharmacol 2003;55:1169-74.
  9. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193:265-75.
  10. Junqueira LC, Bignolas G, Brentani RR. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J 1979;11:447-55.
  11. Woessner JF. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys 1961;93:440-7.
  12. Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol 1978;52:302-10.
  13. Kolios G, Valatas V, Kouroumalis E. Role of Kupffer cells in the pathogenesis of liver disease. World J Gastroenterol 2006; 12:7413-20.
  14. Bataller R, Brenner DA . Liver fibrosis. J Clin Invest 2005;115: 209-18.
  15. Zhu W, Fung PC. The roles played by crucial free radicals like lipid free radicals, nitric oxide, and enzymes NOS and NADPH in CCl(4)-induced acute liver injury of mice. Free Radic Biol Med 2000;29:870-80.
  16. Vendemiale G, Grattagliano I, Caruso ML, Serviddio G, Valentini AM, Pirrelli M, Altomare E. Increased oxidative stress in dimethylnitrosamine-induced liver fibrosis in the rat: effect of N-acetylcysteine and interferon-alpha. Toxicol Appl Pharmacol 2001;175:130-9.
  17. Tsukamoto H, Rippe R, Niemela O, Lin M. Role of oxidative stress in activation of Kupffer cell and Ito cells in liver fibrogenesis. J Gastroenterol Hepatol 1995;10:S50-3.
  18. Parola M, Pinzani M, Casini A, Albano E, Poli G, Gentillini A, Gentillini P, Dianzani MU. Stimulation of lipid peroxidation or 4-hydroxynonenal treatment increases procollagen alpha 1 (I) gene expression in human liver fat-storing cells. Biochem Biophys Res Commun 1993;194:1044-50.
  19. Falchi M, Bertelli A, Lo Scalzo R, Morassut M, Morelli R, Das S, Cui J, Das DK. Comparison of cardioprotective abilities between the flesh and skin of grapes. J Agric Food Chem 2006;54: 6613-22.
  20. Yadav M, Jain S, Bhardwaj A, Nagpal R, Puniya M, Tomar R, Singh V, Parkash O, Prasad GB, Marotta F, Yadav H. Biological and medicinal properties of grapes and their bioactive constituents: an update. J Med Food 2009;12:473-84.
  21. Lee ES, Shin MO, Yoon S, Moon JO. Resveratrol inhibits dimethylnitrosamine-induced hepatic fibrosis in rats. Arch Pharm Res 2010;33:925-32.
  22. Dulundu E, Ozel Y, Topaloglu U, Toklu H, Ercan F, Gedik N, Sener G. Grape seed extract reduces oxidative stress and fibrosis in experimental biliary obstruction. J Gastroenterol Hepatol 2007;22:885-92.
  23. Shin MO, Yoon S, Moon JO. The proanthocyanidins inhibit dimethylnitrosamine-induced liver damage in rats. Arch Pharm Res 2010;33:167-73.
  24. Leifert WR, Abeywardena MY. Cardioprotective actions of grape polyphenols. Nutr Res 2008;28:729-37.
  25. Friedman SL. Mechanisms of hepatic fibrogenesis. Gastroenterology 2008;134:1655-69.
  26. Lazze MC, Pizzala R, Gutiérrez Pecharroman FJ, Gaton Garnica P, Antolin Rodriguez JM, Fabris N, Bianchi L. Grape waste extract obtained by supercritical fluid extraction contains bioactive antioxidant molecules and induces antiproliferative effects in human colon adenocarcinoma cells. J Med Food 2009;12:561-8.
  27. Friedman SL. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem 2000; 275:2247-50.
  28. Chavez E, Reyes-Gordillo K, Segovia J, Shibayama M, Tsutsumi V, Vergara P, Moreno MG, Muriel P. Resveratrol prevents fibrosis, NF-kappaB activation and TGF-beta increases induced by chronic CCl4 treatment in rats. J Appl Toxicol 2008;28:35-43.
  29. Jezequel AM, Mancini R, Rinaldesi ML, Ballardini G, Fallani M, Bianchi F, Orlandi F. Dimethylnitrosamine-induced cirrhosis. Evidence for an immunological mechanism. J Hepatol 1989;8: 42-52.

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