Increased generation of hydrogen peroxide possibly from mitochondrial respiratory chain after UVB irradiation of murine fibroblasts

Abstract

The purpose of this study is to detect the generation of active oxygens in UVB-irradiated murine fibroblasts and to propose new mechanisms. Decreased survival of fibroblasts under UVB irradiation was partially recovered by addition of catalase, DMSO or deferoxamine, suggesting the contribution of several types of active oxygen species. Then we examined the formation of active oxygen species and found that fibroblasts under UVB irradiation generated Superoxide anion radicals (·O − 2), intracellular H 2O 2, and hydroxyl radicals as estimated by the ESR-spin trapping method. Addition of thenoyltrifluoroacetone, which is an inhibitor of the mitochondrial respiratory chain, decreased 29% of the intracellular H 2O 2 levels in UVB-irradiated cells, but allopurinol, which is an inhibitor of xanthine oxidase, had no effect on them. On the basis of these results, we propose a possible mechanism for damage of murine fibroblasts exposed to UVB in terms of generation of active oxygen species. The mitochondrial respiratory chain reaction stimulated by UVB irradiation enhances the generation of ·O − 2, which is in turn dismutated to H 2O 2 and O 2 by Superoxide dismutase. H 2O 2 is then converted to hydroxyl radicals, catalyzed by trace elements such as iron, as suggested by Fenton-like reaction. Thus, hydroxyl radicals with higher reaction rate-constants than those of other active oxygen species to biomolecules are indicated to be responsible for the cytotoxicity in cells under UV irradiation.