Antioxidants vitamin E and C attenuate hepatic fibrosis in biliary-obstructed rats.

AIM To investigate whether antioxidants vitamin E and C can retard development of hepatic fibrosis in the biliary-obstructed rats. METHODS Fifty Wistar albino rats were randomly assigned to 5 groups (10 rats in each). Bile duct was ligated in 40 rats and they were treated as follows: group vitC, vitamin C 10 mg/kg sc daily; group vitE, vitamin E 15 mg/kg sc daily; group vitEC, both of the vitamins; bile duct-ligated (BDL, control) group, physiological saline sc. The fifth group was assigned to sham operation. At the end of fourth week, the rats were decapitated, and hepatic tissue biochemical collagen content and collagen surface area were measured. Hepatic tissue specimens were histopathologically evaluated according to Scheuer system. Serum hyaluronate levels were measured by ELISA method. RESULTS Despite being higher than sham group, hepatic collagen level was significantly decreased in each of the vitC, vitE and vitEC groups (32.7 +/- 1.2, 33.8 +/- 2.9, 36.7 +/- 0.5 mug collagen/mg protein, respectively) compared to BDL (48.3 +/- 0.6 mg collagen/g protein) (P < 0.001 for each vitamin group). Each isolated vitamin C, isolated vitamin E and combined vitamin E/C supplementation prevented the increase in hepatic collagen surface density (7.0% +/- 1.1%, 6.2% +/- 1.7%, 12.3% +/- 2.0%, respectively) compared to BDL (17.4% +/- 5.6%) (P < 0.05 for each). The same beneficial effect of vitamin C, vitamin E and combined vitamin E/C treatment was also observed on the decrease of serum hyaluronate levels compared to BDL group (P < 0.001). The relative liver and spleen weights, serum transaminases, cholestatic enzymes, bilirubins and histopathological inflammation scores were not different between the antioxidant treatment groups and the control. However, fibrosis staging scores were obviously reduced only in the vitamin E/C combination group (vit EC: 2.4 +/- 0.8 vs BDL: 3.1 +/- 0.7; P < 0.05). CONCLUSION Each antioxidant vitamin E, vitamin C and their combination retard hepatic fibrosis in biliary-obstructed rats. Oxidative stress may play a role in the pathogenesis of hepatic fibrosis in secondary biliary cirrhosis.

[1]  G. Poli,et al.  Vitamin E dietary supplementation protects against carbon tetrachloride—induced chronic liver damage and cirrhosis , 1992, Hepatology.

[2]  G. Robino,et al.  Oxidative stress-related molecules and liver fibrosis. , 2001, Journal of hepatology.

[3]  R. Sokol,et al.  Enrichment of rat hepatic organelles by vitamin E administered subcutaneously. , 2004, Free radical biology & medicine.

[4]  G. Poli,et al.  The role of lipid peroxidation in liver damage. , 1987, Chemistry and physics of lipids.

[5]  Yau-Huei Wei,et al.  Oxidative-stress-related changes in the livers of bile-duct-ligated rats. , 2003, Journal of biomedical science.

[6]  P. Hayes,et al.  Serum hyaluronan ‐ a non‐invasive test for diagnosing liver cirrhosis , 2000, European journal of gastroenterology & hepatology.

[7]  D. Schuppan,et al.  Matrix as a modulator of hepatic fibrogenesis. , 2001, Seminars in liver disease.

[8]  A. Gentilini,et al.  Stimulation of lipid peroxidation or 4-hydroxynonenal treatment increases procollagen alpha 1 (I) gene expression in human liver fat-storing cells. , 1993, Biochemical and biophysical research communications.

[9]  P. Scheuer,et al.  Classification of chronic viral hepatitis: a need for reassessment. , 1991, Journal of hepatology.

[10]  D. Schuppan,et al.  Silymarin retards collagen accumulation in early and advanced biliary fibrosis secondary to complete bile duct obliteration in rats , 1997, Hepatology.

[11]  P. Muriel,et al.  Role of glutathione, lipid peroxidation and antioxidants on acute bile-duct obstruction in the rat. , 1999, Biochimica et biophysica acta.

[12]  G. Poli Pathogenesis of liver fibrosis: role of oxidative stress. , 2000, Molecular aspects of medicine.

[13]  Jin-yan Luo,et al.  Estrogen reduces CCL4- induced liver fibrosis in rats. , 2002, World journal of gastroenterology.

[14]  P. Muriel,et al.  Effects of silymarin and vitamins E and C on liver damage induced by prolonged biliary obstruction in the rat. , 2004, Basic & clinical pharmacology & toxicology.

[15]  S. Krähenbühl,et al.  Reduced antioxidative capacity in liver mitochondria from bile duct ligated rats , 1995, Hepatology.

[16]  N. Gedik,et al.  Effects of vitamins E and C supplementation on hepatic glutathione peroxidase activity and tissue injury associated with ethanol ingestion in malnourished rats. , 2006, Current therapeutic research, clinical and experimental.

[17]  D. Brenner,et al.  d-alpha-tocopherol inhibits collagen alpha 1(I) gene expression in cultured human fibroblasts. Modulation of constitutive collagen gene expression by lipid peroxidation. , 1991, The Journal of clinical investigation.

[18]  Harold Anthony Harper,et al.  Harper's review of biochemistry , 1983 .

[19]  S. Friedman Molecular Regulation of Hepatic Fibrosis, an Integrated Cellular Response to Tissue Injury* , 2000, The Journal of Biological Chemistry.

[20]  D. Schuppan,et al.  Antifibrotic effect of silymarin in rat secondary biliary fibrosis is mediated by downregulation of procollagen α1(I) and TIMP-1 , 2001 .

[21]  A. Burt,et al.  In vivo responses of macrophages and perisinusoidal cells to cholestatic liver injury. , 1993, The American journal of pathology.

[22]  I. Dianzani,et al.  Vitamin E dietary supplementation inhibits transforming growth factor β1 gene expression in the rat liver , 1992, FEBS letters.

[23]  M. Rojkind,et al.  A simple micromethod for collagen and total protein determination in formalin-fixed paraffin-embedded sections. , 1985, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[24]  H. Gallo-Torres Obligatory role of bile for the intestinal absorption of vitamin E , 1970, Lipids.

[25]  M. Buck,et al.  Long- and short-termd-α-tocopherol supplementation inhibits liver collagen α1(Ι) gene expression. , 1998, American journal of physiology. Gastrointestinal and liver physiology.

[26]  G. Poli,et al.  Oxidative damage and fibrogenesis. , 1997, Free radical biology & medicine.

[27]  L. Xie,et al.  Effects of dietary supplementation with vitamin E and selenium on rat hepatic stellate cell apoptosis. , 2005, World journal of gastroenterology.

[28]  T. Nakamura,et al.  Inhibitory effect of Y-27632, a ROCK inhibitor, on progression of rat liver fibrosis in association with inactivation of hepatic stellate cells. , 2001, Journal of hepatology.

[29]  J. Kountouras,et al.  Prolonged bile duct obstruction: a new experimental model for cirrhosis in the rat. , 1984, British journal of experimental pathology.

[30]  R. Sokol,et al.  Evidence for involvement of oxygen free radicals in bile acid toxicity to isolated rat hepatocytes , 1993, Hepatology.

[31]  B. Fleshler,et al.  The Liver: Biology and Pathobiology , 1983 .

[32]  R. Roth,et al.  Bile and bile salts potentiate superoxide anion release from activated, rat peritoneal neutrophils. , 1988, Toxicology and applied pharmacology.

[33]  C. Trautwein,et al.  Stimulation of collagen α1(I) gene expression is associated with lipid peroxidation in hepatocellular injury: A link to tissue fibrosis? , 1994, Hepatology.

[34]  P. Montilla,et al.  Melatonin versus vitamin E as protective treatment against oxidative stress after extra‐hepatic bile duct ligation in rats , 2001, Journal of pineal research.

[35]  P. Hayashi,et al.  Vitamin E and vitamin C treatment improves fibrosis in patients with nonalcoholic steatohepatitis , 2003, American Journal of Gastroenterology.

[36]  P. Flecknell,et al.  Experimental and surgical technique in the rat. , 1980 .

[37]  E. Ah-Sing,et al.  Antioxidant defenses in the bile duct-ligated rat. , 1992, Gastroenterology.

[38]  Rong-hua Liu,et al.  Influence factors of serum fibrosis markers in liver fibrosis. , 2003, World journal of gastroenterology.

[39]  K. Hickman Fat-Soluble Vitamins , 1943 .

[40]  P. Scheuer,et al.  LIVER BIOPSY INTERPRETATION , 1973, The Ulster Medical Journal.