Effects of Vitamins C and E on Hepatic Drug Metabolizing Function in Nypoxia/Reoxygenation

저산소 및 산소재도입시 vitamin C와 E가 간장 약물대사 기능에 미치는 영향

  • Published : 2000.06.01

Abstract

Liver isolated from 18 hours fasted rats was subjected to $N_2$hypoxia (for 45 min) followed by reoxygenation (for 30 min). The perfusion medium used was Krebs-Henseleit bicarbonate buffer (pH 7.4, $37^{\circ}C$). Vitamin C (0.5 mM) and trolox C (0.5 mM), soluble vitamin E analog, were added to perfusate. Lactate dehydrogenase (LDH), total glutathione, oxidized glutathione, lipid peroxide and drug-metabolizing enzymes were measured. After hypoxia LDH significantly increased but this increase was attenuated by vitamin C and combination of vitamin C and E. Total glutathione and oxidized glutathione in perfusate markedly increased during hypoxia and this increase was inhibited by vitamins C, E and its combination. Similarly; oxidized glutathione and lipid peroxide in liver tissue increased after hypoxia and reoxygenation and this increase was inhibited by vitamin I and combination of vitamin C and E. Hepatic drug metabolizing function (phase I, II) were suppressed during hypoxia but improved during reoxygenation. While vitamins C and E only increased glucuronidation, the combination of vitamin C and E increased the oxidation, glucuronidation and sulfation. Our findings suggest that vitamins C and E synergistically ameliorates hepatocellular damage as indicated by abnormalities in drug metabolizing function during hypoxia/reoxygenation and that this protection is in major part, caused by decreased oxidative stress.

Keywords

References

  1. Acta. Physiol. Scand. v.548 Ischemia-reperfusion injury:role of oxygen-derived free radicals Granger. D.N.;Hollwarth. M.E.;Parks. D.A.
  2. In Free radicals from basic science to medicine Pharmaceutical intervention for the prevention of post-ischemic reperfusion injury Mayumi. T.;Schiller. H.J.;Bukley .G.B.;G. Poli.(ed.)
  3. Hepatology v.21 Alteration of xanthine oxidase activity in sinusoidal endothelial cells and morphological changes of Kuffer cells in hypoxic and rexygenated rat liver. Angermuller. S.;Schunk. M.;Kusterer. K.
  4. Am. J. physiol. v.G355 Neutrophil and Kupffer cell-induced oxidant stress and ischemia-reperfusion injury in rat liver Jaeschke. H.;Farhood. A.
  5. Transplant. Proc. v.27 no.1 Inactivation of Kupffer cells minimizes reperfusion injury in fat-loaded livers from ethanol-treated rats Zhong. Z.;qu. W.;Connor. H.D.;Thurman. R.G.
  6. Hepatology v.3 Trolox protects rat hepatocytes against oxyradical damage and the ischemic rat liver from reperfusion injury Wu. T.W.;Hashimoto. N.;Au. J.X.;Wu. J.;Carry. D.
  7. Surgery v.99 Role of free radicals in ischemic rat liver cell injury. Prevention of damage by α-tocopherol administration Marubayashi. S.;Dohi. K.;Ochi. K.;Kawasaki. T.
  8. J. Pharmacol. Exp. Ther. v.236 New, simple models to evaluate zone-specific damage due to hypoxia in the perfused rat liver: time course and effect of nutritional state Bradford. B.U.;Marotto. M.;Lemasters. J.J.;Thurman. R.G.
  9. Transplantation v.58 The effects of aging and nutritional state on hypoxiareoxygenation injury in the perfused rat liver. Le Couteur. D.G.;Rivory. L.P.;Pond. S.M.
  10. Bichem. Pharmacol. v.38 Enhancement of acute ethanol hepatotoxicity under conditions of low oxygen supply and ischemia/reperfusion Younes. M.;Wagner. H.;Strubelt. O.
  11. Anal. Biochem. v.74 Enzymatic assay for glutathione Brehe. J.E.;Burch. H.B.
  12. Anal. Biochem. v.106 Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinyl pyridine Griffith. O.W.
  13. Int. J. Vitam. Nutr. Res. v.46 Effect of vitamin E deficiency on the level of superoxide dismutase, gluatathione peroxidase, catalase and lipid peroxidation Masugi. F.;Nakamura. T.
  14. Anal. Biochem. v.72 A rapid and sensitive method of the quantitation of microgram quantities of protein utilizing the principle of protein dye binding Bradford. M.M.
  15. Chem. Biol. Interactions v.61 Functional relationship between intital oxidation of 7-ethoxy-coumarin and subsequent conjugation of 7-hydroxy-coumarin in isolated perfused rat livers Cha. Y.N.;Dong. M.S.;Hong. S.S.
  16. Am. J. Physiol. v.253 Fructose prevents hypoxic cell death in liver Anundi. I.;King. J.;Owen. D.A.;Schneider. H.;Lemasters. J.J.;Thurman. R.G.
  17. J. Clin. Invest. v.87 Enhanced activity of the free radical producing enzyme xantine oxidase in hypoxic rat liver Brass. C.A.;Narciso. J.;Gollan. J.L.
  18. Am. J. Physiol. v.255 Role of xanthine oxidase and granulocytes in ischemia-reperfusion injury Granger. D.N.
  19. J. Clin. Invest. v.81 Reactive oxygen species during ischemia-reflow injury in isolated perfused rat liver Jaschke. H.;Smith. C.V.;Mitchell. J.R.
  20. Biochem. Biophys. Acta v.835 Antioxidant and co-antioxidant activity of vitamin C. The effect of vitamin C, either alone or in the presence of vitamin E of a water soluble vitamin E analogue upon the peroxidation of agueous multilameilar phospholipid liposomes Doba. T.;Burton. G.W.;Ingold. K.W.
  21. Biochem. Biophys. Acta. v.664 The cooperative interaction between vitamin E and vitamin C in suppression of peroxidation of membrane phospholioids Leung. H.W.;Vang. M.J.;Mavis. R.D.
  22. Biochem. Biophys. Res. Com. v.150 Hypoxic damage generates reactive oxygen species in isolated perfused rat liver Jaeschke. H.;Smith. C.V.;Mitchell. J.R.
  23. In Handbook of free radicals and antioxidants in biomedicine The physiological functions of gluathione Borg. D.C.;Schaich. K.M.;Jaime Miquel.(ed.)
  24. Biochem. Cell Biol. v.69 Trolox and ascorbate: are they synergistic in protecting liver cells in vitro and in vivo? Zeng. L.H.;Wu. J.;Carey. D.;Wu. T.W.
  25. Hepatology v.15 Effect of α-tocopherol on hepatic mixed function oxidase in hepatic ischemia/reperfusion Lee. S.M.;Clemens. M.G.
  26. Hoppeseylers Z. Physiol. Chem. v.353 The O-dealkylation of 7-ethoxycoumarin by liver microsomes Ullrich. W.;Weber. P.
  27. Drug Metab. Dispos v.24 [14C]7-ethoxycoumarin metabolism by precision-cut rat hepatic slices Simon. E.b.;Victoria. E.T.;Cesario. O.T.;Lim. H.K.;Joh. K.;Samuel. F.S.;Joann. S.
  28. Hepatology v.20 Mitochondrial dysfunction during anoxia/reoxygenation injury of liver sinusoidal endothelial cells Fujii. Y.;Johnson. M.E.;Gores. G.J.
  29. Am. J. Clin. Nutr. v.34 An increase in vitamin E requirement induced by high supplementation of vitamin C in rat. Chen. L.H.