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Hepatic response to the oxidative stress induced by E. coli endotoxin: Glutathione as an index of the acute phase during the endotoxic shock

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Abstract

Reactive oxygen species are important mediators of cellular damage during endotoxic shock. In order to investigate the hepatic response to the oxidative stress induced by endotoxin, hepatic and plasma glutathione (total, GSH and GSSG), GSSG/GSH ratio as well as Mn-superoxide dismutase and catalase activities were determined during the acute and recovery phases of reversible endotoxic shock in the rat. A significant increase in liver and plasma total glutathione content was observed 5 h after endotoxin treatment (acute phase), followed by a diminution of these parameters below control values at 48 h (recovery phase). The significant increases of GSSG levels and GSSG/GSH ratio are indicative of oxidative stress occurring during the acute phase. Liver Mn-SOD activity showed a similar time dependency as the GSSG/GSH ratio; however, a marked decrease in the liver catalase activity was observed during the process. These results indicate the participation of liver glutathione in the response to endotoxin and the possible use of plasma glutathione levels and GSSG/GSH ratio as indicators of the acute phase during the endotoxic process. (Mol Cell Biochem 159: 115-121, 1996)

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References

  1. Arthur MJP, Bentley IS, Tanner AR, Saunders PK, Millward-Sadler GH, Wright R: Oxygen derived free radicals promote hepatic injury in the rat. Gastroenterol 89: 1114–1122, 1985

    Google Scholar 

  2. Bautista AP, Mészáros K, Bojta J, Spitzer JJ: Superoxide anion generation in the liver during the early stage of endotoxemia in rats. J Leukocyte Biol 48: 123–128, 1990

    Google Scholar 

  3. Bautista AP, Spitzer JJ: Superoxide anion generation by in situ perfused rat liver: effect of in vivo endotoxin. Am J Physiol 259: G907-G912, 1990

    Google Scholar 

  4. Comporti M: Biology of disease-lipid peroxidation and cellular damage in toxic liver injury. Lab Invest 53: 599–623, 1985

    Google Scholar 

  5. Fantone JC, Ward PA: Role of oxygen-derived free radicals and metabolites in leukocyte-dependent inflammatory reaction. Am J Physiol 107: 397–401, 1982

    Google Scholar 

  6. Freeman B, Crapo JD: Free radicals and tissue injury. Lab Invest 47: 412–426, 1982

    Google Scholar 

  7. Ogawa R, Morita T, Kunimoto F, Fujita T: Changes in hepatic lipoperoxide concentration in endotoxemic rats. Circ Shock 9: 369–374, 1982

    Google Scholar 

  8. Sevanian A, Kim E: Phospholipase A2 dependent release of fatty acids from peroxidized membranes. Free Rad Biol Med 1: 263–271, 1985

    Google Scholar 

  9. Portolés MT, Ainaga MJ, Pagani R: The induction of lipid peroxidation by E. coli lipopolysaccharide on rat hepatocytes as an important factor in the etiology of endotoxic liver damage. Biochim Biophys Acta 1158: 287–292, 1993

    Google Scholar 

  10. Geller BL, Winge DR: Subcellular distribution of superoxide dismutases in rat liver. Meth Enzymol 105: 105–114, 1984

    Google Scholar 

  11. Kawaguchi T, Takeyuasu A, Matsunobu K, Uda T, Ishizawa M, Suzuki K, Nishiura T, Ishikawa M, Taniguchi N: Stimulation of Mn-superoxide dismutase expression by tumor necrosis factor-α: Quantitative determination of Mn-SOD protein levels in TNF-resistant and sensitive cells by ELISA. Biochem Biophys Res Commun 171: 1378–1386, 1990

    Google Scholar 

  12. Masuda A, Longo DL, Kobayashi Y, Appella E, Oppenheim JJ, Matsushima K: Induction of mitochondrial manganese superoxide dismutase by interleukin 1. FASEB J 2: 3087–3091, 1988

    Google Scholar 

  13. Shaffer JB, Treanor CP, Del Vecchio PJ: Expression of bovine and mouse endothelial cell antioxidant enzymes following TNF-α exposure. Free Rad Biol Med 8: 497–502, 1990

    Google Scholar 

  14. Visner GA, Dougall WC, Wilson JM, Burr IM, Nick HS: Regulation of manganese superoxide dismutase by lipopolysaccharide, interleukin-1, and tumor necrosis factor. Role in the acute inflammatory response. J Biol Chem 265: 2856–2864, 1990

    Google Scholar 

  15. Visner GA, Block ER, Burr IM, Nick HS: Regulation of manganese superoxide dismutase in porcine pulmonary artery endothelial cells. Am J Physiol 260: L444–449, 1991

    Google Scholar 

  16. Wong GHW, Goeddel DV: Induction of manganous superoxide dismutase by tumor necrosis factor: Possible protective mechanism. Science 242: 941–944, 1988

    Google Scholar 

  17. Tyler DD: Polarographic assay and intracellular distribution of superoxide dismutase in rat liver. Biochem J 147: 493–504, 1975

    Google Scholar 

  18. Dhaunsi GS, Gulati S, Singh AK, Orak JK, Asaymama K, Singh I: Demonstration of Cu,Zn-superoxide dismutase in rat liver peroxisomes. J Biol Chem 267: 6870–6873, 1992

    Google Scholar 

  19. Keller GA, Warner TG, Steiner KS, Hallewell RA: Cu,Zn superoxide dismutase is a peroxisomal enzyme in human fibroblast and hepatoma cells. Proc Natl Acad Sci USA 88: 7381–7385, 1991

    Google Scholar 

  20. 20.Aebi H: Catalase in vitro. Meth Enzymol 105: 121–126, 1984

    Google Scholar 

  21. Lazarow PB, Fujiki Y: Biogenesis of peroxisomes. Ann Rev Cell Biol 1: 489–530, 1985

    Google Scholar 

  22. Splitz DR, Adams DT, Sherman CM, Roberts RJ: Mechanisms of cellular resistance to hydrogen peroxide, hyperoxia and 4-hydrogen peroxide, hyperoxia and 4-hydroxy-2-nonenal toxicity: the significance of increased catalase activity in H2O2-resistant fibroblast. Arch Biochem Biophys 292: 221–227, 1992

    Google Scholar 

  23. Guillemette J, Marion M, Denizeau F, Fournier M, Brousseau P: Characterization of the in vitro hepatocyte model for toxicological evaluation: repeated growth stimulation and glutathione response. Biochem Cell Biol 71: 7–13, 1993

    Google Scholar 

  24. Kaplowitz N, Aw TK, Ookhtens M: The regulation of hepatic glutathione. Ann Rev Pharmacol Toxicol 25: 715–744, 1985

    Google Scholar 

  25. Meister A, Anderson ME: Glutathione. Ann Rev Biochem 52: 711–760, 1983

    Google Scholar 

  26. Ketterer B, Coles B, Meyer DJ: The role of glutathione in detoxification. Environ Health Perspect 49: 59–69, 1983

    Google Scholar 

  27. Lauterburg BH, Adams JD, Mitchell JR: Hepatic glutathione homeostasis in the rat: efflux accounts for glutathione turnover. Hepatology 4: 586–590, 1984

    Google Scholar 

  28. Lu SC, Garcia-Ruiz C, Kuhlenkamp J, Ookhtens M, Salas-Prato M, Kaplowitz N: Hormonal regulation of glutathione efflux. J Biol Chem 265: 16088–16095, 1990

    Google Scholar 

  29. Sies H, Graf P: Hepatic thiol and glutathione efflux under the influence of vasopressin, phenylephrine and adrenaline. Biochem J 226: 545–549, 1985

    Google Scholar 

  30. Westphal O, Lüderitz OS, Bister F: Extraction of bacteria with phenol water. Z Naturf (b) 7: 148–155, 1952

    Google Scholar 

  31. Tietze F: Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem 27: 502–522, 1969

    Google Scholar 

  32. Griffith OW: Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinyl pyridine. Anal Biochem 106: 207–212, 1980

    Google Scholar 

  33. Marklund S, Marklund G: Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47: 469, 1974

    Google Scholar 

  34. Lowry OH, Rosebrough NJ, Farr AL, Randall R: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  35. Beer RF, Sizer WI: A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195: 133–140, 1952

    Google Scholar 

  36. Lück H. In: H.U. Bergmeyer (ed.). Methods of Enzymatic Analysis. Verlag Chemie, Weinheim-Bergsstrasse, Germany, 1965, pp 885–894

    Google Scholar 

  37. Garcia R, Diaz-Laviada I, Bosch MA, Abarca S, Portolés MT, Risco C, Ainaga MJ, Pagani R, Municio AM, Escalona J: Morphological studies of cytotoxic lesions in reversible endotoxic shock. Histol Histopath 4: 43–48, 1990

    Google Scholar 

  38. Bosch MA, Garcia R, Pagani R, Portolés MT, Diaz-Laviada I, Abarca S, Ainaga MJ, Risco C, Municio, AM: Induction of reversible shock by Escherichia coli lipopolysaccharide in rats. Changes in serum and cell membrane parameters. Br J Exp Pathol 69: 805–812, 1988

    Google Scholar 

  39. Shimizu M, Morita S: Effects of feeding and fasting on hepatolobular distribution of glutathione and cadmium-induced hepatotoxicity. Toxicol 75: 97–107, 1992

    Google Scholar 

  40. Ookhtens M, Mittur AV: Developmental changes in plasma thioldisulfidetumover in rats: a multicompartmental approach. Am J Physiol 267: R415-R425, 1994

    Google Scholar 

  41. Mathison JC, Ulevitch RJ: The clearance tissue distribution and cellular localization of intravenously injected lipopolysaccharide in rabbits. J Immunol 123: 2133–2143, 1979

    Google Scholar 

  42. Nolan JP: Endotoxin, reticuloendothelial function and liver injury. Hepatology 1: 458–465, 1981

    Google Scholar 

  43. Demling R, La Londe C, Daryani R, Zhu DG, Knox J, Youn YK: Relationship between the lung and systemic response to endotoxin. Comparison of physiologic change and the degree of lipid peroxidation. Circ Shock 34: 364–370, 1991

    Google Scholar 

  44. Kunimoto F, Morita R, Ogawa R, Fugita T: Inhibition of peroxidation improves survival rate of endotoxemic rats. Circ Shock 21: 15–22, 1987

    Google Scholar 

  45. Powell RJ, Machiedo GW, Rush BF Jr, Dikdan GS: Effect of oxygen-free radical scavengers on survival in sepsis. Am Surg 57: 86–88, 1991

    Google Scholar 

  46. Romero C, Bosch MA: Effect of dimethylthiourea in phosphathidyl choline biosynthesis by rat lung during reversible endotoxic shock. Mol Cell Biochem 129: 1–7, 1993

    Google Scholar 

  47. Schneider J, Matthiesen T: The prostacyclin analogue taprostene and recombinant human superoxide dismutase increase the permanent survival rate of endotoxemic rats. Life Sci 46: 1421–1426, 1990

    Google Scholar 

  48. Portolés MT, Arahuetes RM, Pagani R: Intracellular calcium alterations and free radical formation evaluated by flow cytometry in endotoxin-treated rat liver Kupffer and endothelial cells. Eur J Cell Biol 65: 200–205, 1994

    Google Scholar 

  49. Portolés MT, Ainaga MJ, Municio AM, Pagani R: Intracellular calcium and pH alterations induced by Escherichia coli endotoxin in rat hepatocytes. Biochim Biophys Acta 1092: 1–6, 1991

    Google Scholar 

  50. Jaeschke H: Vascular oxidant stress and hepatic ischemic-reperfusion injury. Free Rad Res Commun 12–13: 737–743, 1991

    Google Scholar 

  51. Jaeschke H: Enhanced sinusoidal glutathione efflux during endotoxin-induced stress in vivo. Am J Physiol: 263, G60-G68, 1992

    Google Scholar 

  52. Wohaieb SA, Godin DV: Starvation-related alterations in free radical defense mechanisms in rats. Diabetes 36: 167–173, 1987

    Google Scholar 

  53. Kaplan JH, Groves, JN: Liver and blood cell catalase activity of tumor-bearing mice. Cancer Res 32: 1190–1194, 1972

    Google Scholar 

  54. Gulati S, Singh AK, Irazu C: Ischemia-reperfusion injury: biochemical alterations in peroxisomes of rat kidney. Arch Biochem Biophys 295: 90–100, 1992

    Google Scholar 

  55. Pagani R, Portolés MT, Bosch MA, Diaz-Laviada I, Municio AM: Direct and mediated Escherichia coli lipopolysaccharide action in primary hepatocyte cultures. Eur J Cell Biol 43: 243–246, 1987

    Google Scholar 

  56. Bray RC, Cockle SA, Fielden EM, Roberts PB, Rotilio G, Calabrese L: Reduction and inactivation of superoxide dismutase by hydrogen peroxide. Biochem J 139: 43–48, 1974

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Faculty of Chemistry, Universidad Computense, 28040, Madrid, Spain

    M. Teresa Portolés, Myriam Catalá, Adolfo Antón & Raffaella Pagani

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  1. M. Teresa Portolés
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  2. Myriam Catalá
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  3. Adolfo Antón
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  4. Raffaella Pagani
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Portolés, M.T., Catalá, M., Antón, A. et al. Hepatic response to the oxidative stress induced by E. coli endotoxin: Glutathione as an index of the acute phase during the endotoxic shock. Molecular and Cellular Biochemistry 159, 115–121 (1996). https://doi.org/10.1007/BF00420913

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