Effects of Chronic Alcohol Feeding and 2-Acetylaminofluorene Treatment on Microsomal Cytochrome P-450 and Glutathione Dependent Enzymes Activities in Rat Liver

만성 알코올 섭취시 2-Acetylaminofluorene 투여가 흰쥐간 Cytochrome P-450 및 Glutathione 이용 효소계 활성에 미치는 영향

  • 김정희 (서울여자대학교 영양학과) ;
  • 최옥희 (서울여자대학교 영양학과) ;
  • 윤혜진 (서울여자대학교 영양학과)
  • Published : 1995.12.01

Abstract

This study was done to investigate the effects of chronic ethanol feeding on hepatic microsomal cytochrome system, lipid peroxidation and peroxide metabolizing enzyme activities in 2-acetylaminofluorene(2-AAF) treated rats. Male Sprague-Dawley rats, weighing 120~125g, were pair-fed liquid diets containing 35% of total calories either as ethanol or isocaloric carbohydrates for 6 weeks. After 4 weeks of experimental diet feeding, 2-AAF(100mg/kg body weight) was injected twice a week intraperitoneally. Both weight and percent liver weight per body weight were significantly changed by ethanol feeding. Hepatic microsomal lipid peroxide value and the activities of glutathione(GSH) peroxidase and GSH reductase were not changed by either ethanol or 2-AAF treatment. However the analysis of cytochrome systems showed that both ethanol and 2-AAF increased cytochrome P-450 and bs contents although cytochrome P-450 content was moe affected by 2-AAF while cytochrome b5 content by ethanol. Cytosolic GSH S-transferase activity, which is often elevated during chemical carcinogenesis, also significantly increased by either ethanol feeding or 2-AAF treatment. Overall values for the cytochrome contents and GSH S-transferase activities were highest in 2-AAF treated rats fed ethanol. These results might support the hypothesis that the increase in liver cancer risk associated with chronic ethanol consumption might be due to, at least in part, enhancement of carcinogen bioactivation by ethanol.

본 연구에서는 액체식이로 알코올을 열량의 35%로 6주간 섭취시킨 흰주의 간조직내 지질과산화물과 glutathione 이용효소계의 활성도 및 cytochrome P-450에 미치는 영향을 살펴보고 아울러 간암의 발암원으로 알려진 2-AAF를 투여하여 이들의 상호효과를 조사한 결과 다음과 같은 결과를 얻었다. 1. 체중, 간무게, 그리고 체중에 대한 간무게의 알코올에 대한 효과는 유의적인 차이를 보여 체중은 알코올에 대한 효과는 유의적인 차이를 보여 체중은 알코올 섭취군이 유의적으로 감소하였고 간무게 및 체중에 대한 간무게는 알코올군의 유의적으로 증가하였다. 2. Microsome의 지질과산화물 함량 및 cytosol의 glutathione peroxidase, glutathione reductase 활성도는 알코올과 2-AAF 투여시 모두 유의적인 차이를 보이지 않았으나, cytosol의 glutathione S-transferase 활성도는 알코올과 2-AAF에 의해서 모두 유의적으로 증가하였고 알코올 섭추와 함께 투여시 GST 활성도가 가장 많이 증가하였다. 3. Microsome의 cytochrome P-450 및 cytochrome b5 함량에 대한 알코올 효과는 cytochrome P-450 함량을 증가시키는 경향이 있고 cytochrome b5는 유의적인 증가를 보여 주었으며 2-AAF 투여 역시 cytochrome P-450의 유의적인 증가를 유도하였다. 따라서 알코올 섭취와 2-AAF 함께 투여 시 cytochrome P-450의 함량이 대조군의 약 2.2배 정도 증가하였으며 cytochrome b5 함량이 1.7배로 높이 증가하였다. 이는 2-AAF가 cytochrome P-450을 유도하여 자신의 대사를 촉진시키며 알코올의 섭취 또한 2-AAF의 hydroxylation을 증가시킬 수 있는 것으로 생각된다. 이상의 결과에서 과량의 알코올을 만성적으로 섭취시 간조직내의 microsome의 MFO system에 영향을 미쳐서 발암물질의 생체 활성화를 촉진시킬 수 있고 또한 GST의 활성도를 증가시키므로 어느 정도 발암과정에 영향을 미치는 것으로 생각된다.

Keywords

References

  1. Medical and nutritional complications of alcoholism : Mechanisms and management Alohol and liver Lieber,C.S.;Leo,M.A.;Lieber,C.S.(ed.)
  2. Hepatology v.6 Ethanol induced liver fibrosis in rats fed high fat diets Tsukamoto,H.;Towner,S.J.
  3. Gastroenterology v.106 Alcohol and the liver : 1994 update Lieber,C.S.
  4. J. Clin. Invest. v.40 The effect of ethanol on fatty acid metabolism : Stimulation of hepatic fatty acid synthesis in vitro Lieber,C.S.;Schimid,R.
  5. Hepatology v.6 Alcohol and cancer Lieber,C.S.;Garro,A.;Leo,M.A.;Mak,K.M.;Wornor,T.
  6. Glutathione, alcohol and hepatotoxicity Nutrition and the origins of disease Kaplowitz,N.;Fernandez-Checa,J.;Ookhtens,M.;Halsted,C.H.(ed.);Rucker,R.B.(ed.)
  7. Biochem. Biophys. Res. Commun. v.143 Acetaldehyde-mediated hepatic lipid peroxidation : Role of superoxide and ferritin Shaw,S.;Jayatilleke,E.
  8. Pharmcol. Rev. v.24 The membrane actions of anesthetics and tranquilizers Seeman,P.
  9. J. Nutr. v.118 Differential effects of reinoids and chronic ethanol consumption on membranes in rats Kim,C.I.;Leo,M.A.;Lowe,N.;Lieber,C.S.
  10. Research advances in alcohol and drug problem v.7 Phsico-chemical interactions between alcohol and biological membranes Michaeli,E.K.;Michael,M.L.;Smart,R.G.(ed.);Glaser,H.(ed.);Israel,Y.(ed.);Kalant,J.M.(ed.);Popham,A.(ed.);Schmidt,A.(ed.)
  11. Lab. Invest. v.14 Hepatic changes associated with chronic alcoholism in rats Porta,E.A.;Hartroft,W.S.;de la lglesia,F.A.
  12. Arch. Biochem. Biophys. v.165 Effect of chronic ethanol treatment on mitochondrial functions : Damage to coupling site I Cederbaum,A.I.;Lieber,C.S.;Rubin,E.
  13. J. Biol. Chem. v.254 Effects of chronic ethanol intoxication on the respiratory chain of rat liver submitochondrial particles Thayer,W.S.;Rubin,E.
  14. Lab. Invest. v.42 The effect of ethanol on the temperature dependence of respiration and ATPase activities of rats liver mitochondria Rottenburg,H.;Robertson,D.;Rubin,E.
  15. Arch. Biochem. Biophys. v.183 The influence of chronic ethanol feeding to rats on the integrity of liver mitochondrial membrane as assessed with the $Mg^ {3+}$-stimulated ATPase enzyme Hosein,E.A.;Hofmann,I.;Linder,E.
  16. Ann. Rev. Biochem. v.44 Chemical carcinogenesis Heidelberger,C.
  17. Alcoholism : Clin. Exp. Res. v.10 Ethanol and carcinogenesis of the alimentary tract Seitz,H.K.;Simanowski,U.A.
  18. Nutr. Rev. v.36 Oxygen activation in the metabolism of lipids, drugs and carcinogens Coon,M.J.
  19. Alcohol & Alcoholism v.24 Liquid diet technique of ethanol administration 1989 update Lieber,C.S.;DeCarli,L.M.
  20. Methods in enzymology v.10 Isolation of liver and kidney mitochondria Johnson,D.;Lardy,H.;Estabrook,R.W.(ed.);Pullman,ME.(ed.)
  21. Plant. Physiol. v.86 Cytochemical localization of ATPase activity in roots Katz,D.B.;Sussman,M.R.;Mierzwa,R.J.;Evert,R.F.
  22. Methods in enzymology v.4 Lowry,O.H.;Sidney,P.C.(ed.);Nathan,O.(ed.)
  23. J. Biol. Chem. v.193 Protein measurment with the folin phenol reagent Lowry,O.H.;Rosebrough,N.J.;Randall,R.J.
  24. Can. J. Biochem. Phys. v.37 A rapid method of total lipid extraction and purification Bligh,E.G.;Dyer,N.J.
  25. Clin. Chem. v.20 Enzymatic determination of total serum cholesterol Allain,C.C.;Poon,L.S.;Chan,C.S.G.;Richmond,W.;Fu,P.C.
  26. Lipids v.5 Two dimensional thin layer chromatographic separation of polar lipids and determination of phospholipids by phosphorus Rouser,G.;Fleischer,S.;Yamamoto,A.
  27. J. Lipid. Res. v.23 Seperation of phospholipids and individual molecular species of phospholipids by high-performance liquid chromatography Patton,G.M.;Fasulo,J.M.;Robins,S.J.
  28. Anal. Chem. v.38 Rapid preparation of fatty acid esters from lipids for gas chromatographic analysis Metcalfe,L.D.;Schmitz,A.A.;Pelka,J.R.
  29. Alcoholism : Clin & Exp, Res. v.6 The feeding of alcohol in liquid diests : Two decade of applications and 1982 update Lieber,C.S.;DeCarli,L.M.
  30. Normal alcohols. Biochemistry v.15 Interactions between anesthetics and lipid mixtures Lee,A.G.
  31. Biochem. Biophys. Acta v.797 Decreased cytochrome oxiddase activity in hepatic mitochondria after chronic ethanol consumption and the possible role of decreased cytochrome a-3 content and changes in phospholipids Arai,M.;Gordon,E.R.;Leber,C.S.
  32. Biochem. Biophys. Acta v.712 Ethanol consumption on the phospholipid composition of rat liver microsomes and mitochondria Cunningham,C.C.;Filus,S.;Bottenus,R.E.;Spach,P.I.
  33. Science v.216 Lack of correlation between hepatic mitochondrial membrane structure and functions in ethanol-fed rats Gordon,E.R.;Arai,M.;Lieber,C.S.
  34. Korean J. Nutr. v.26 Effects of different dietary oils in hepatic mitochondria lipid composition, adenosine nucleotide translocase and ATPase activities in carcinogen treated rats Lee,M.S.;Kim,J.H.
  35. Lipids v.8 Effects of ethanol on membrane lipids, III. Quantitative changes in lipids and fatty acid composition of nonpolar and polar lipids of mouse total liver mitochondria and microsome following ethanol feeding Miceli,J.N.;Ferrell,W.J.
  36. Hepatology v.8 Effects of vitamin A and ethanol on liver plasma membrane fluidity Kim,C.I.;Leo,M.A.;Lowe,N.;Lieber,C.S.
  37. J. Nutr. v.114 Effect of dietary fat level and polyundaturated fatty acid content on the phospholipid composition of rat cardiac mitochondrial membranes and mitochondrial ATAase activity Robblee,N.M.;Clandinin,M.T.
  38. I. Hepatic mitochondria. Mech. Aging Rev. v.46 Age-related changes in respiration coupled to phosphorylation Kim,J.H.;Woldgiorgis,G.;Elson,C.E.;Shrago,E.