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Non-enzymatic and enzymatic lipid peroxidation of microsomes and nuclei obtained from rat liver

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Abstract

The present study investigates in a experimental system in vitrothe relationship between the non-enzymatic (ascorbate-Fe2+) and enzymatic (NADPH) lipid peroxidation in rat liver microsomes and nuclei. Chemiluminescence was measured as cpm/mg protein during 180 min at 37 °C. Approximately 50–55% of the fatty acids located in rat liver microsomes and nuclei are polyunsaturated with a prevalence of C18:2 n6 and C20:4 n6. The values of total light emission during the non-enzymatic and enzymatic lipid peroxidation were highest in microsomes than in nuclei. A significant decrease of C20:4 n6 and C22:6 n3 in rat liver microsomes and nuclei was observed during the lipid ascorbate-Fe2+-dependent peroxidation, whereas a significant decrease of C20:4 n6 in rat liver microsomes was observed during enzymatic lipid peroxidation. Over the time course studies, analysis of chemiluminescence in microsomes and nuclei demonstrated that the lipid peroxidation in the presence of ascorbate-Fe2+ reach a maximum at 50 and 30 min, respectively, whereas in the presence of NADPH it reachs a maximum at 20 min in both organelles. In liver microsomes and nuclei the peroxidizability index (pi) which indicates the degree of vulnerability to degradation of a selected membrane showed statistically significant differences between control versus ascorbate-Fe2+ when microsomes or nuclei were compared. Our results indicate that non-enzymatic (ascorbate-Fe2+) and enzymatic (NADPH) lipid peroxidation are operative in rat liver microsomes and nuclei but the sensitivities of both organelles to lipid peroxidation evidenced by chemiluminescence was greater in the presence of ascorbate-Fe2+ when compared with NADPH. (Mol Cell Biochem 265: 1–9, 2004)

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References

  1. Proteggente AR, Rehman A, Halliwell B, Rice-Evans CA: Potential problems of ascorbate and iron supplementation: Pro-oxidant effect in vivo? Biochem Biophys Res Commun 277: 535–540, 2000.

    Google Scholar 

  2. Welch KD, Davis TZ, Van Eden ME, Aust SD: Deleterious iron-mediated oxidation of biomolecules. Free Radic Biol Med 32: 577–583, 2002.

    Google Scholar 

  3. Girotti AW: Mechanisms of lipid peroxidation. J Free Radic Biol Med 1: 87–95, 1985.

    Google Scholar 

  4. Tadolini B, Hakim G: The mechanism of iron(III) stimulation of lipid peroxidation. Free Radic Res 25: 221–227, 1996.

    Google Scholar 

  5. Pamplona R, Portero-Otin M, Riba D, Ruiz C, Prat J, Bellmunt MJ, Barja G: Mitochondrial membrane peroxidizability index is inversely related to maximum life span in mammals. J Lipid Res 39: 1989–1994, 1998.

    Google Scholar 

  6. Ozgova S, Hermanek J, Gut I: Different antioxidant effects of polyphenols on lipid peroxidation and hydroxyl radicals in the NADPH-, Fe-ascorbate-and Fe-microsomal systems. Biochem Pharmacol 66: 1127–1137, 2003.

    Google Scholar 

  7. Dmitriev LF: Activity of key enzymes in microsomal and mitochondrial membranes depends on the redox reactions involving lipid radicals. Membr Cell Biol 14: 649–662, 2001.

    Google Scholar 

  8. Hsieh RJ, Kinsella JE: Oxidation of polyunsaturated fatty acids: Mech-anisms, products, and inhibition with emphasis on fish. Adv Food Nutr Res 33: 233–241, 1989.

    Google Scholar 

  9. Halliwell B, Gutteridge JMC: Role of free radicals and catalytic metal ions in human disease: An overview. Meth Enzymol 186: 1–85, 1990.

    Google Scholar 

  10. Palmer S: Antioxidant vitamins and cancer risk. Nutrition 10: 433–434, 1994.

    Google Scholar 

  11. Aruoma OI: Nutrition and health aspects of free radicals and antioxidants. Fd Chem Toxic 32: 671–683, 1994.

    Google Scholar 

  12. Bonorden WR, Pariza MW: Antioxidant nutrients and protection from free radicals. In: F.N. Kotsonis (ed). Nutritional Toxicology.Raven Press, New York, 1994, pp 19–45.

    Google Scholar 

  13. Vladimirov Yu A, Olenev VI, Suslova TB, Cheremisina ZP: Lipid peroxidation in mitochondrial membrane. Adv Lipid Res 17: 173–249, 1980.

    Google Scholar 

  14. Wright JR, Colby HD, Miles PR: Cytosolic factors which affects microsomal lipid peroxidation in lung and liver. Arch Biochem Biophys 206: 296–304, 1981.

    Google Scholar 

  15. Galleano M, Puntarulo S: Hepatic chemiluminescence and lipid peroxidation in mild iron overload. Toxicology 76: 27–38, 1992

    Google Scholar 

  16. Puntarulo S, Cederbaum AI: Stimulation of microsomal chemilumines cence by ferritin. Biochim Biophys Acta 1157: 1–8, 1993.

    Google Scholar 

  17. Gavazza M, Catala A: The effect of á-tocopherol on the lipid peroxida tion of mitochondria and microsomes obtained from rat liver and testis. Mol Cell Biochem 225: 121–128, 2001.

    Google Scholar 

  18. Marmunti M, Catalá A:Non-Enzymatic lipid peroxidation of rat liver nuclei and chromatin fractions. Int J Biochem Cell Biol 30: 967–972, 1998.

    Google Scholar 

  19. Tangen O, Jonsson J, Orrenius S: Isolation of rat liver microsomes by gel filtration. Anal Biochem 54: 597–603, 1973.

    Google Scholar 

  20. Yasmineh WG, Yunis JJ: Localization of mouse satellite DNA in constitutive heterochromatin. Exp Cell Res 59: 69–75, 1970.

    Google Scholar 

  21. Yasmineh WG, Yunis JJ: Isolation of mammalian heterochromatin and euchromatin. In: D.M. Prescott (ed). Methods in Cell Biology,vol. VIII, Academic Press, New York, 1974, pp 151–177.

    Google Scholar 

  22. Wright JR, Rumbaugh RC, Colby HD, Miles PR: The rela tionship between chemiluminescence and lipid peroxidation in rat hepatic microsomes. Arch Biochem Biophys 192: 344–351, 1979.

    Google Scholar 

  23. Tadolini B, Cabrini L, Menna C, Pinna GG, Hakim G: Iron[III] stimu lation of lipid hydroperoxide dependent lipid peroxidation. Free Radic Res 27: 563–576, 1997.

    Google Scholar 

  24. Folch J, Lees N, Sloane Stanley GA: A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226: 497–509, 1957.

    Google Scholar 

  25. Pirozhkov SV, Eskelson CD, Watson RR, Hunter GC, Piotrowski JJ, Bernhard V: Effect of chronic consumption of ethanol and vitamin E on fatty acids composition and lipid peroxidation in rat heart tissue. Alcohol 9: 329–334, 1992.

    Google Scholar 

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

    Google Scholar 

  27. Di Mascio P, Murphy ME, Sies H: Antioxidants defense systems: The role of carotenoids, tocopherols and thiols. Ann J Clin Nutr 53: 194S–200S, 1991.

    Google Scholar 

  28. Dix TA, Aikens J: Mechanims and biological relevance of lipid peroxidation initiation. Chem Res Toxicol 6: 2–18, 1993.

    Google Scholar 

  29. Monahan FJ, Gray JI, Asghar A, Haug A, Strasburg GM, Buckley DJ, Morrissey PA: Influence of diet on lipid oxidation and membrane struc ture in porcine muscle microsomes. J Agric Food Chem 42: 59–63, 1994.

    Google Scholar 

  30. Vaca EC, Wilthelm J, Harms-Rindahl M: Studies on lipid peroxidation in rat liver nuclei and isolated nuclear membranes. Biochim Biophys Acta 958: 375–387, 1988.

    Google Scholar 

  31. Zanetti R, Catalá A:Changes in n-6 and n-3 polyunsaturated fatty acids during lipid peroxidation of mitochondria obtained from rat liver and several brain regions: Effect of á-tocopherol. Prostaglandins Leukot Essent Fatty Acids 62: 379–385, 2000.

    Google Scholar 

  32. Mlakar A, Spiteller G: Distinction between enzymic and nonenzymic lipid peroxidation. J Chromatogr A 743: 293–300, 1996.

    Google Scholar 

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Authors and Affiliations

  1. Cátedra de Bioquímica, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, La Plata, Argentina

    Mónica Marmunti, Mariana Gavazza & Angel Catalá

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  1. Mónica Marmunti
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  2. Mariana Gavazza
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  3. Angel Catalá
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Marmunti, M., Gavazza, M. & Catalá, A. Non-enzymatic and enzymatic lipid peroxidation of microsomes and nuclei obtained from rat liver. Mol Cell Biochem 265, 1–9 (2004). https://doi.org/10.1023/B:MCBI.0000044302.59193.00

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