Use of the Chemiluminigenic Probes Luminol and Lucigenin for the Detection of Active Oxygen Species in Hepatic Microsomes and in Intact Hepatocytes
During microsomal NADPH oxidation, reduced oxygen species are produced which must be considered a normal by-product of the NADPH-cytochrome P-450 reductase/cytochrome P-450 system (Gillette, Brodie and LaDu, 1957; Hildebrandt, Speck and Roots,1973; Estabrook, Kawano, Werringloer et al., 1979; Cederbaum and Dicker, 1983). Formation of such reactive oxygen molecules is enhanced in the presence of many monooxygenase substrates (Hildebrandt and Roots, 1975; Hildebrandt, IIeinemeyer and Roots, 1982) and may contribute to the toxicity of foreign compounds undergoing microsomal biotransformation ( Farber and Gerson, 1984 ). It has been suggested that superoxide anion radicals are released during autoxidation of the oxycytochrome P-450 complex and are subsequently dismutated to yield hydrogen peroxide ( Ullrich and Kuthan, 1980 ); hydroxyl radical formation may then result from hydrogen peroxide cleavage in the presence of superoxide anion by a Haber-Weiss reaction ( Haber and Weiss, 1934 ).
KeywordsButylate Hydroxy Toluene Propyl Gallate Butylate Hydroxyanisole Propyl Gallate Photon Yield
Unable to display preview. Download preview PDF.
- Allen, R.C., 1982, Biochemiexcitation: Chemiluminescence and the study of biological oxygenation reactions, in: “ Chemical and Biological Generation of Excited States”, Academic Press, New York, p. 309.Google Scholar
- Cederbaum, A.I., and Dicker, E., 1983, Inhibition of microsomal oxidation of alcohols and of hydroxyl radical-scavenging agents by the ironchelating agent desferrioxamin, Biochem.J., 210: 10.Google Scholar
- Cummings, S.W., and Prough, R.A., 1983, Butylated hydroxyanisolestimulated NADPH oxidase activity in rat liver microsomal fractions, J.biol.Chem., 258: 1 2315.Google Scholar
- Farber, J.L., and Gerson, R.J., 1984, Mechanisms of cell injury with hepatotoxic chemicals, Pharmacol.Rev. 36: 715.Google Scholar
- Fridovich, I, 1976, Oxygen Radicals, hydrogen peroxide, and oxygen toxicity, in: “Free Radicals in Biology”, W.A.Pryor, ed., Academic Press, New York, p. 239.Google Scholar
- Orrenius, S., Thor, H., Eklöw, L., Moldeus, P., and Jones, D.P., 1982, Drug-stimulated H202 formation in hepatocytes. Possible toxicological implications, in: “Biological Reactive Intermediates”, R.Snyder, D.V.Parke, J.J.Kocsis, D.Jollow, C.G.Gibson, C.M.Witmer, eds., Plenum Press, New York and London, p. 395.Google Scholar
- Powis, G., Svingen, B.A., Appel, P., 1982, Factors affecting the intracellular generation of free radicals from quinones, in: “Biological Reactive Intermediates”, R.Snyder, D.V.Parke, J.J.Kocsis, D.J.Jollow, C.G.Gibson, C.M.Witmer, eds., Plenum Press, New York and London, P. 349.Google Scholar
- Ullrich, V., and Kuthan, H., 1980, Autoxidation and uncoupling in microsomal monooxygenations, in: “Biochemistry, Biophysics and Regulation of Cytochrome P-450, J.Gustafsson, J. Carlstedt-Duke, A.Mode, J.Rafter, eds., Elsevier, Amsterdam, p. 267.Google Scholar