Advertisement

Cell Biology and Toxicology

, Volume 19, Issue 6, pp 355–366 | Cite as

Adaptation of subcellular glutathione detoxification system to stress conditions in choline-deficient diet induced rat fatty liver

  • I. GrattaglianoEmail author
  • P. Caraceni
  • P. Portincasa
  • M. Domenicali
  • V.O. Palmieri
  • F Trevisani
  • M Bernardi
  • G. Palasciano
Article

Abstract

The response of fatty liver to stress conditions (t-butyl hydroperoxide [t-BH] or 36 h of fasting) was investigated by assessing intracellular glutathione (GSH) compartmentation and redox status, GSH peroxidase (GSH-Px) and reductase (GSSG-Rx) activities, lipid peroxidation (TBARs) and serum ALT levels in rats on a choline-deficient diet. Baseline cytosolic GSH was similar between fatty and normal livers, while the mitochondrial GSH content was significantly lower in fatty livers. With the except of cytosolic GSH-Px activity, steatosis was associated with significantly higher GSH-related enzymes activities. Liver TBARs and serum ALT levels were also higher. Administration of t-BH significantly decreased the concentration of cytosolic GSH, increased GSSG levels in all the compartments, and increased TBARs levels in cytosol and mitochondria and serum ALT; all these alterations were more marked in rats with fatty liver. Fasting decreased the concentration of GSH in all the compartments both in normal and fatty livers, increased GSSG, TBARs and ALT levels, and decreased by 50% the activities of GSH-related enzymes. Administration of diethylmaleimide (DEM) resulted in cytosolic and microsomal GSH pool depletion. Administration of t-BH to DEM-treated rats further affected cytosolic GSH and enhanced ALT levels, whereas the application of fasting to GSH depleted rats mainly altered the mitochondrial GSH system, especially in fatty livers. This study shows that fatty livers have a weak compensation of hepatic GSH regulation, which fails under stress conditions, thus increasing the fatty liver's susceptibility to oxidative damage. Differences emerge among subcellular compartments which point to differential adaptation of these organelles to fatty degeneration.

choline-deficient diet fasting fatty liver glutathione depletion microsomes mitochondria 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adams JD, Lauterburg BH, Mitchell JR. Plasma glutathione and glutathione disulfide in the rat: regulation and response to oxidative stress. J Pharmacol Exp Ther. 1983;227:749–54.Google Scholar
  2. Alpers DH, Sabesin SM, White HM. Fatty liver: biochemical and clinical aspects. In: Shiff L, Shiff E, eds. Disease of the liver. Philadelphia: Lippincott; 1993:825–55.Google Scholar
  3. Arrigo AP. Gene expression and the thiol redox state. Free Radic Biol Med. 1999;27:936–44.Google Scholar
  4. Berson A, de Beco V, Letteron P, et al. Steato-hepatitis inducing drugs cause mitochondrial dysfunction and lipid peroxidation in rat hepatocytes. Gastroenterology. 1998;114:764–74.Google Scholar
  5. Brodie AE, Reed DJ. Glutathione disulfide reduction in tumor mitochondria after t-butyl hydroperoxide treatment. Chem Biol Interact. 1992;84:125–32.Google Scholar
  6. Caraceni P, Nardo B, Domenicali M, et al. Ischemia-reperfusion injury in the rat fatty liver: role of nutritional status. Hepatology. 1999;29:1139–46.Google Scholar
  7. Carlberg I, Mannervik B. Glutathione reductase. Methods Enzymol. 1985;113:484–90.Google Scholar
  8. Chiba T, Takahashi S, Sato N, Ishii S, Kikuchi K. Fas-mediated apoptosis is modulated by intracellular glutathione in human T cells. Eur J Immunol. 1996;26:1164–9.Google Scholar
  9. Clottes E, Burchell A. Three thiol groups are important for the activity of the liver micrososmal glucose-6-phosphatase system. Unusual behavior one thiol located in the glucose-6-phosphate translocase. J Biol Chem. 1998;273:19391–7.Google Scholar
  10. DeLeve LD, Kaplowitz N. Glutathione metabolism and its role in hepatotoxicity. Pharmacol Ther. 1991;52:287–305.Google Scholar
  11. Dianzani MU, Muzio G, Biocca ME, Canuto RA. Lipid peroxidation in fatty liver induced by caffeine in rats. Intl J Tiss React. 1991;13:79–85.Google Scholar
  12. Fernandez-Checa JC, Hirano T, Tsukamoto H, Kaplowitz N. Mitochondrial glutathione depletion in alcoholic liver disease. Alcohol. 1993;10:469–75.Google Scholar
  13. Fernandez-Checa JC, Jian-R YI, Garcia-Ruiz C, Okhtens M, Kaplowitz N. Plasma membrane and mitochondrial transport of hepatic reduced glutathione. Semin Liv Dis. 1996;16:147–58.Google Scholar
  14. Flohé L, Gunzler WA. Assays of glutathione peroxidase. Methods Enzymol. 1984;105:114–21.Google Scholar
  15. Folch J, Lees M, Sloane-Stanley GH. A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem. 1957;226:497–502.Google Scholar
  16. Ginn-Pease ME, Whisler RL. Optimal NF kappa B mediated transcriptional responses in Jurkat T cells exposed to oxidative stress are dependent on intracellular glutathione and costimulatory signals. Biochem Biophys Res Commun. 1996;226:695–702.Google Scholar
  17. Grattagliano I, Vendemiale G, Lauterburg BH. Reperfusion injury of the liver: role of mitochondria and protection by glutathione ester. J Surg Res. 1999;86:2–8.Google Scholar
  18. Grattagliano I, Vendemiale G, Caraceni P, et al. Antioxidant defense is impaired by starvation in rat fatty livers induced by a choline-depleted diet. J Nutr. 2000;130:2131–6.Google Scholar
  19. Griffith OW. Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem. 1980;106:207–12.Google Scholar
  20. Guerrieri F, Vendemiale G, Grattagliano I, Cocco T, Pellecchia G, Altomare E. Mitochondrial oxidative alterations following partial hepatectomy. Free Radic Biol Med. 1999;26:34–41.Google Scholar
  21. Guerrieri F, Vendemiale G, Turturro N, et al. Alteration of mitochondrial F0F1 ATP syntahse during aging. Possible involvement of oxygen free radicals. Ann NY Acad Sci. 1996;786:62–71.Google Scholar
  22. Gupta M, Dobashi K, Greene EL, Orak JK, Singh I. Studies on hepatic injury and antioxidant enzyme activities in rat subcellular organelles following in vivo ischemia and reperfusion. Mol Cell Biochem. 1997;176:337–47.Google Scholar
  23. Hakamada K, Sasaki M, Takahashi K, Umehara Y, Konn M. Sinusoidal flow block after warm ischemia in rats with diet-induced fatty liver. J Surg Res. 1997;70:12–20.Google Scholar
  24. Hammond CL, Lee TK, Ballatori N. Novel roles for glutathione in gene expression, cell death, and membrane transport of organic solutes. J Hepatol. 2001;34:946–54.Google Scholar
  25. Hoppel CJ, Dimarco D, Tandler B. Riboflavin and rat hepatic cell structure and function. J Biol Chem. 1979;254:4164–70.Google Scholar
  26. Huang ZZ, Chen C, Zeng Z, et al. Mechanism and significance of increased glutathione level in human hepatocellular carcinoma and liver regeneration. FASEB J. 2001;15:19–21.Google Scholar
  27. Hwang C, Sinskey AJ, Lodish HF. Oxidized redox state of glutathione in the endoplasmic reticulum. Science. 1992;257:1496–502.Google Scholar
  28. Lauterburg BH, Velez ME. Glutathione deficiency in alcoholics: risk factor for paracetamol hepatotoxicity. Gut. 1988;29:1153–7.Google Scholar
  29. Martensson J, Meister A. Glutathione deficiency decreases tissue ascorbate levels in newborn rats; ascorbate spares glutathione and protects. Proc Natl Acad Sci USA. 1991;89:4656–60.Google Scholar
  30. Meister A. Glutathione deficiency produced by inhibition of its synthesis, and its reversal; applications in research and therapy. Pharmacol Ther. 1991;51:155–94.Google Scholar
  31. Mitchell JR, Jollow JD, Potter WZ, Gillette JR, Brodie BB. Acetaminophen-induced hepatic necrosis. IV. Protective role of glutathione. J Pharmacol Exp Ther. 1973;187:211–27.Google Scholar
  32. Mosialou E, Ekstrom G, Adang AE, Morgenstern R. Evidence that rat liver microsomal glutathione transferase is responsible for glutathione dependent protection against lipid peroxidation. Biochem Pharmacol. 1993;45:1645–51.Google Scholar
  33. Nagai H, Matsumaru K, Feng G, Kaplowitz N. Reduced glutathione depletion causes necrosis and sensitization to tumor necrosis factor-α-induced apoptosis in cultured mouse hepatocytes. Hepatology. 2002;36:55–64.Google Scholar
  34. Nardo B, Grattagliano I, Domenicali M, et al. Mitochondrial oxidative injury in rat fatty livers exposed to warm ischemia-reperfusion. Transplant Proceed. 2000;32:51.Google Scholar
  35. Olafsdottir K, Reed DJ. Retention of oxidizied glutathione by isolated rat liver mitochondria during hydroperoxide treatment. Biochim Biophys Acta. 1988;964:377–82.Google Scholar
  36. Palamanda JR, Kehrer JP. Inhibition of protein carbonyl formation and lipid peroxidation by glutathione in rat liver microsomes. Arch Biochem Biophys. 1992;293:103–9.Google Scholar
  37. Reed DJ, Orrenius S. The role of methionine in glutathione biosynthesis by isolated hepatocytes. Biochem Biophys Res Commun. 1977;77:1257–64.Google Scholar
  38. Robertson G, Lequercq I, Farrell GC. Non-alcoholic steatosis and steato-hepatitis. II. Cytochrome P-450 enzymes and oxidative stress. Am J Physiol Gastrointest Liver Physiol. 2001;281:G1135–G42.Google Scholar
  39. Russmann S, Junker E, Lauterburg BH. Remethylation and transsulfuration of methionine in cirrhosis: studies with L-[H3-methyl-1C]methionine. Hepatology. 2002;36:1190–6.Google Scholar
  40. Santori G, Domenicotti C, Belloccio A, et al. Effects of acute glutathione depletion induced by L-buthionine-(S,R)-sulfoximine on rat liver glucose-6-phosphatase activity. Res Commun Mol Pathol Pharmacol. 1997;98:165–78.Google Scholar
  41. Schafer FQ, Buettner GR. Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radic Biol Med. 2001;30:1191–212.Google Scholar
  42. Sies H. Glutathione and its role in cellular functions. Free Radic Biol Med. 1999;27:916–21.Google Scholar
  43. Sies H, Akerboom TPM. Glutathione disulfide (GSSG) efflux from cells and tissues. Methods Enzymol. 1984;105:445–51.Google Scholar
  44. Slater T, Sawyer B. The stimulatory effect of carbon tetrachloride and other halogenoalkanes on peroxidative reactions in rat liver fraction in vitro. Biochem J. 1971;123:805–14.Google Scholar
  45. Soltys K, Dikdan G, Koneru B. Oxidative stress in fatty livers of obese zucker rats: rapid amelioration and improved tolerance to warm ischemia with tocopherol. Hepatology. 2001;34:13–8.Google Scholar
  46. Spolarics Z, Meyenhofer M. Augmented resistance to oxidative stress in fatty rat livers induced by a short-term sucrose-rich diet. Biochim Biophys Acta. 2000;1487:190–200.Google Scholar
  47. Tietze F. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: application to mammalian blood and other tissues. Anal Biochem. 1969;27:502–22.Google Scholar
  48. Vendemiale G, Guerrieri F, Grattagliano I, Didonna D, Muolo L, Altomare E. Mitochondrial oxidative phosphorylation and intracellular glutathione compartmentation during rat liver regeneration. Hepatology. 1995;21:1450–4.Google Scholar
  49. Vendemiale G, Grattagliano I, Altomare E, Turturro N, Guerrieri F. Effect of acetaminophen administration on hepatic glutathione compartmentation and mitochondrial energy metabolism in the rat. Biochem Pharmacol 1996;52:1147–54.Google Scholar
  50. Vendemiale G, Grattagliano I, Caraceni P, et al. Mitochondrial oxidative injury and energy metabolism alteration in rat fatty liver: effect of the nutritional status. Hepatology. 2001;33:808–15.Google Scholar
  51. Yang CF, Shen, HM, Ong CN. Ebselen induces apoptosis in HepG(2) cells through rapid depletion of intracellular thiols. Arch Biochem Biophys. 2000;374:142–52.Google Scholar
  52. Yang S, Zhu H, Li Y, et al. Mitochondrial adaptation to obesity-related oxidant stress. Arch Biochim Biophys. 2000;378:259–68.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • I. Grattagliano
    • 1
    Email author
  • P. Caraceni
    • 2
  • P. Portincasa
    • 1
  • M. Domenicali
    • 2
  • V.O. Palmieri
    • 1
  • F Trevisani
    • 2
  • M Bernardi
    • 2
  • G. Palasciano
    • 1
  1. 1.Section of Internal Medicine and Department of Internal Medicine and Public Medicine (DIMIMP)University Medical School of BariItaly
  2. 2.Section of Internal Medicine and Department of Internal Medicine, Cardioangiology and HepatologyUniversity Medical School of BolognaItaly

Personalised recommendations