Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8993135 | Drug Metabolism and Pharmacokinetics | 2005 | 10 Pages |
Abstract
Previously, we reported that singlet oxygen (1O2) was involved in rat liver microsomal P450-dependent substrate oxygenations in such reactions as p-hydroxylation of aniline, O-deethylation of 7-ethoxycoumarin, Ï- and (Ï-1)-hydroxylations of lauric acid, O-demethylation of p-nitroanisole, and N-demethylation of aminopyrine. In order to confirm the generality of 1O2 involvement, we have further investigated which kinds of reactive oxygen species (ROS) are formed during P450-dependent substrate oxygenation in microsomes. We examined CYP2E1-dependent hydroxylation of p-nitrophenol in rat liver microsomes in the presence of some ROS scavengers, because CYP2E1 has been reported to predominantly generate ROS in the hepatic microsomes and to relate with the oxidative stress in the body. The addition of 1O2 quenchers, β-carotene, suppressed the hydroxylation of p-nitrophenol. Furthermore, a nonspecific P450 inhibitor, SKF525A, and a ferric chelator, deferoxamine, both suppressed the hydroxylation. No other ROS scavengers such as superoxide dismutase (SOD), catalase, or mannitol altered the reaction. 1O2 was detectable during the reaction in the microsomes as measured by an electron spin resonance (ESR) spin-trapping method when 2,2,6,6-tetramethyl-4-piperidone (TMPD) was used as a spin-trapping reagent. The 1O2 was quenched by additions of β-carotene, p-nitrophenol, and SKF525A. The reactivity of p-nitrophenol and 1O2 correlated linearly with its hydrox- ylation rate in the microsomes. On the basis of these results, we conclude that 1O2 contributes to the p-nitrophenol hydroxylation in rat liver microsomes, by adding a new example of 1O2 involvement in the CYP2E1-dependent substrate oxygenations.
Keywords
p-Nitrophenol hydroxylationDeferoxamineDMPOTMPD4-Oxo-TEMPOESR1O2CYP2E1P450NADPHDFOCAT2,2,6,6-Tetramethyl-4-piperidoneDMSOROS·OHHydrogen peroxideSinglet oxygen (1O2)Singlet oxygenDimethylsulfoxideSuperoxide anion radicalHydroxyl radicalElectron spin resonanceSODSuperoxide dismutaseCytochrome P450NADHRate constantNicotinamide adenine dinucleotide reduced formnicotinamide adenine dinucleotide phosphate reduced formH2O2CatalaseReactive oxygen species
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Authors
Seiko Hayashi, Hiroyuki Yasui, Hiromu Sakurai,