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Glutathione and antioxidants protect microsomes against lipid peroxidation and enzyme inactivation

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Lipids

Abstract

The study investigated the relationship between lipid peroxidation and enzyme inactivation in rat hepatic microsomes and whether prior inactivation of aldehyde dehydrogenase (ALDH) exacerbated inactivation of other enzymes. In microsomes incubated with 2.5 μM iron as ferric sulfate and 50 μM ascorbate, ALDH, glucose-6-phosphate (G6Pase) and cytochrome P450 (Cyt-P450) levels decreased rapidly and concurrently with increased levels of thiobarbituric acid-reactive substances. Microsomal glutathioneS-transferase and nicotinamide adenine dinucleotide phosphate-cytochromec reductase were little affected during 1 hr of incubation. Addition of reduced glutathione partially protected, andN,N′-diphenyl-p-phenylenediamine and butylated hydroxytoluene completely protected microsomes against inactivation of ALDH, G6Pase and Cyt-P450, as well as lipid peroxidation induced by iron and ascorbate. ALDH was more susceptible than G6Pase to inactivation by iron and ascorbate, and was thus an excellent marker for oxidative stress. Inhibition of ALDH by cyanamide injection of rats exacerbated the inactivation of G6Pase in microsomes incubated with 0.1 mM, but not 25 μM 4-hydroxynonenal (4-HN). 4-HN did not stimulate lipid peroxidation. Thus, 4-HN may play a minor role in microsomal enzyme inactivation. In contrast, lipid, peroxyl radicals play an important role in microsomal enzyme inactivation, as evidenced by the prevention of both lipid peroxidation and enzyme inactivation by chain-breaking antioxidants.

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Abbreviations

Aa:

ascorbic acid

ALDH:

aldehyde dehydrogenase(s)

BHT:

butylated hydroxytoluene

Cyt-c-Rx:

cytochromec reductase

Cyt-P450:

cytochrome P450

DPPD:

N,N′-diphenyl-p-phenylenediamine

EDTA:

disodium ethylenedinitrilotetraacetate

GSH:

reduced glutathione

G6Pase:

glucose-6-phosphase

GSHS-Tr:

glutathioneS-transferase

4-HN:

4-hydroxynonenal

TBARS:

thiobarbituric acid-reactive substances

References

  1. Tappel, A.L. (1973)Fed. Proc. 32, 1870–1875.

    PubMed  CAS  Google Scholar 

  2. Davies, K.J.A. (1988) inCellular Antioxidant Defense Mechanisms (Chow, C.K., ed.) Vol. 2, pp. 27–40, CRC Press, Boca Raton.

    Google Scholar 

  3. Poli, G., Biasi, C.F., Leonarduzzi, G., Comoglio, A., Albano, E., and Dianzani, M.U. (1989)Adv. Biosci. 76, 73–81.

    Google Scholar 

  4. Esterbauer, H. (1985) inFree Radicals in Liver Injury (Poli, G., Cheeseman, K.H., Dianzani, M.U., and Slater, T.F., eds.), pp. 29–47, IRL Press, Oxford.

    Google Scholar 

  5. Brattin, W.J., Glende, Jr., E.A., and Recknagel, R.O. (1985)J. Free Radicals Biol. Med. 1, 27–38.

    Article  CAS  Google Scholar 

  6. Eckl, P., and Esterbauer, H. (1989)Adv. Biosciences 76, 141–157.

    Google Scholar 

  7. Witz, G. (1989)Free Radical Biol. Med. 7, 333–349.

    Article  CAS  Google Scholar 

  8. Haenen, G.R.M.M., Tsoi, J.N.L.T.T., Vermeulen, N.P.E., Timmerman, H., and Bast, A. (1987)Arch. Biochem. Biophys. 259, 449–456.

    Article  PubMed  CAS  Google Scholar 

  9. Koivula, T., and Koivusalo, M. (1975)Biochim. Biophys. Acta 397, 9–23.

    PubMed  CAS  Google Scholar 

  10. Nakayasu, M., Mihara, K., and Sato, R. (1978)Biochem. Biophys. Res. Commun. 83, 697–703.

    Article  PubMed  CAS  Google Scholar 

  11. Hjelle, J.J., Grubbs, J.H., Beer, D.G., and Petersen, D.R. (1981)J. Pharmacol. Exp. Therapeu. 219, 821–826.

    CAS  Google Scholar 

  12. Mitchell, D.Y., and Petersen, D.R. (1987)Toxicol. Appl. Pharmacol. 87, 403–410.

    Article  PubMed  CAS  Google Scholar 

  13. Mitchell, D.Y., and Petersen, D.R. (1989)Arch. Biochem. Biophys. 269, 11–17.

    Article  PubMed  CAS  Google Scholar 

  14. Hu, M.-L., Frankel, E.N., Leibovitz, B.E., and Tappel, A.L. (1989)J. Nutr. 119, 1574–1582.

    PubMed  CAS  Google Scholar 

  15. Beuge, J.A., and Aust, S.D. (1978)Methods Enzymol. 52, 301–310.

    Google Scholar 

  16. Harper, A.E. (1963)Methods of Enzymatic Analysis (Bergmeyer, H.U., ed.), pp. 788–792, Academic Press, New York.

    Google Scholar 

  17. Masters, B.S.S., Williams, Jr., C.H., and Kamin, H. (1967)Methods Enzymol 13, 565–574.

    Article  Google Scholar 

  18. Omura, T., and Sato, R. (1964)J. Biol. Chem. 239, 2370–2378.

    PubMed  CAS  Google Scholar 

  19. Habig, W.H., Pabst, M.J., and Jakoby, W.B. (1974)J. Biol. Chem. 249, 7130–7139.

    PubMed  CAS  Google Scholar 

  20. Thomas, P.D., and Poznansky, M.J. (1990)J. Biol. Chem. 265, 2684–2691.

    PubMed  CAS  Google Scholar 

  21. Itoh, F., Minamide, Y., Horie, T., and Awazu, S. (1989)Lipids 24, 905–908.

    PubMed  CAS  Google Scholar 

  22. Nordlie, R.C., and Sukalski, K.A. (1985) inThe Enzymes of Biological Membranes (Martonosi, A.N., ed.) Vol. 2, pp. 349–398, Plenum Press, New York.

    Google Scholar 

  23. Dillard, C.J., Hu, M.-L., and Tappel A.L. (1991)Free Radical Biol. Med. 10, 51–60.

    Article  CAS  Google Scholar 

  24. Morgenstern, R., and Depirre, W. (1983)Eur. J. Biochem. 134, 591–597.

    Article  PubMed  CAS  Google Scholar 

  25. Ando, M., and Tappel, A.L. (1985)Chem.-Biol. Interact. 55, 317–326.

    Article  PubMed  CAS  Google Scholar 

  26. Speth, M., Baake, N., and Schulze, H.-U. (1989)Arch. Biochem. Biophys. 275, 202–214.

    Article  PubMed  CAS  Google Scholar 

  27. Woenkhaus, C., Bieber, E., and Jeck R. (1987) inEnzymology and Molecular Biology of Carbonyl Metabolism (Weiner, H., and Flynn, T.G., eds.), pp. 53–65, Alan R. Liss, New York.

    Google Scholar 

  28. Lame, M.W., and Segall, H.J. (1987)Chem.-Biol. Interact. 62, 59–75.

    Article  PubMed  CAS  Google Scholar 

  29. Benedetti, A., Casini, A.F., Ferrali, M., and Comporti, M. (1979)Biochem. Pharmacol. 28, 2909–2918.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Food Science and Technology, University of California, 109 Food Science and Technology Building, 95616, Davis, CA

    Miao-Lin Hu & Al L. Tappel

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  1. Miao-Lin Hu
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  2. Al L. Tappel
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Hu, ML., Tappel, A.L. Glutathione and antioxidants protect microsomes against lipid peroxidation and enzyme inactivation. Lipids 27, 42–45 (1992). https://doi.org/10.1007/BF02537057

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  • DOI: https://doi.org/10.1007/BF02537057

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