Abstract
In the present study, we examined the effect of the intraperitoneal administration of vitamin E (100 mg/kg weight/24 h) on ascorbate (0.4 mM) induced lipid peroxidation of rat liver microsomes . We also analyzed the effect of hepatic cytosolic proteins on this process. The results indicate that the ascorbate induced light emission was 76% lower in microsomes (1 mg protein) obtained from vitamin E treated animals when compared with controls. In the presence of cytosolic protein (1 mg) the chemiluminescence of control microsomes diminished 55.8 and 59.5% when cytosol from controls and treated animals was used, respectively. The chemiluminescence of vitamin E microsomes diminished 25.03 and 22.08% when both types of cytosol were added to the medium. Dialyzed or treated at 70°C cytosol was also able to inhibit the lipid peroxidation of either control or vitamin E rat liver microsomes. By means of gas chromatography we analyzed the fatty acid composition of native and peroxidated microsomes from both animal groups. The peroxidation affected principally arachidonic acid and its diminution was more evident in the control microsomes than in the microsomes from the vitamin E treated group. By HPLC we analyzed the vitamin E content in all subcellular fractions employed. In microsomes from the vitamin E-group, the content of vitamin was 11 times higher than in the control ones (0.678 ± 0.1038 vs. 0.062 ± 0.0045 μg α-tocopherol/mg protein, respectively), while levels in the cytosol from the vitamin E-group were only 2 times higher than in the control cytosol (0.057 ± 0.0051 vs. 0.025 ± 0.0015 μg α-tocopherol/mg protein, respectively).
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References
McCay PB: Vitamin E: Interactions with free radicals and ascorbate. Annu Rev Nutr 5: 323–340, 1985
Duthie GG, McPhail DB, Morrice PC, Arthur JR: Dietary vitamin E decreases ESR signal intensity in hepatic microsomal preparations from malignant hyperthermia susceptible pigs. Free Rad Res Commun 14: 133–137, 1991
Dutta-Roy AK, Leishman DJ, Gordon MJ, Campbell FM, Duthie GG: Identification of a low molecular mass (14,2 Kda) α-tocopherol-binding protein in the cytosol of rat liver and heart. Biochem Biophys Res Commun 196: 1108–1112, 1993
Tangen O, Jonsson J, Orrenius S: Isolation of rat liver microsomes by gel filtration. Anal Biochem 54: 597–603, 1973
Wright JR, Rumbaugh RC, Colby HD, Miles PR: The relation between chemiluminescence and lipid peroxidation in rat hepatic microsomes. Arch Biochem Biophys 192: 344–351, 1979
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
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with Folin phenol reagent. J Biol Chem 193: 265–275, 1951
Malnoc A, Heuzelin Y, Stanley JC: Phospholipid fatty acid composition and vitamin E level in retina of obese (sa/sa) and lean Zucker rats. Biochim Biophys Acta 1212: 119–124, 1994
Castignani GL: An HPLC method for the simultaneous determination of retinol and α-tocopherol in plasma or serum. Meth Enzymol 123: 215–219, 1986
Tappel AL: Lipid peroxidation damage to cell components. Fed Proc 32: 1870–1874, 1973
Burton GW, Ingold KU: Autooxidation of biological molecules. 1. The antioxidant activity of vitamin E and related chain-breaking phenolic antioxidants in vitro. J Am Chem Soc 103: 6472–6477, 1981
Burton GW, Joyce A, Ingold KU: Is vitamin E the only lipid soluble chain breaking antioxidant in human blood plasma and erythrocites membranes? Arch Biochem Biophys 221: 281–290, 1983
Liebler DC, Baker PF, Kaysen KL: Oxidation of vitamin E: Evidence for competing autoxidation and peroxyl radical trapping reactions of the tocopheroxyl radical. J Am Oil Chem Soc 112: 6995–7000, 1990
Borello S, Minotti G, Palombini G, Grattagliano A, Galeotti T: Superoxide dependent lipid peroxidation and vitamin E of microsomes from hepatomas with different growth rate. Arch Biochem Biophys 238: 588–595, 1985
Ross D, Moldeus P: Antioxidant defense systems and oxidative stress. In: C Vigo-Pelfrey (ed). Membrane Lipid Oxidation. Boca Raton, Fl: CRC, 1991, pp 151–170
Lucy JA: Functional and structural aspects of biological membranes: A suggested structural role of vitamin E in the control of membrane permeability. Ann New York Acad Sci 203: 4–11, 1972
Molenaar Y, Hulstaert CE, Vos J: Membrane characteristics in vitamin E deficiency and the assessment of vitamin E status. Proc Nutr Soc 32: 249–254, 1973
Antonenkov V, Sies H: Ascorbate-dependent capacity of dialyzed rat liver cytosol to prevent nonenzymatic lipid peroxidation. Biol Chem Hoppe-Seyler 373: 1111–1116, 1992
Antonenkov V, Sies H: Some basic properties of ascorbate dependent antioxidative defence factor from rat liver cytosol. Chem Biol Interactions 94:169–181, 1995
Fukuzawa K, Chida H, Tokumura A, Tsukatani H: Antioxidative effect of α-tocopherol incorporation into lecithin liposomes on ascorbic acid-Fe2+-induced lipid peroxidation. Arch Biochem Biophys 206: 173–180, 1981
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Zanetti, R., Catalá, A. Ascorbate-Fe2+ lipid-peroxidation of rat liver microsomes: Effect of vitamin E and cytosolic proteins. Mol Cell Biochem 183, 49–54 (1998). https://doi.org/10.1023/A:1006843932049
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DOI: https://doi.org/10.1023/A:1006843932049