Time course of changes in antioxidant enzymes in human skin fibroblasts after UVA irradiation

Examining damage (inactivation) of antioxidant enzymes in the cells and the pattern of recovery after a single UV exposure might be a useful method for analyzing the mechanisms of chronic UV irradiation, because chronic UV irradiation means repeated single exposures. We irradiated human skin fibroblasts with a single exposure to UVA (1, 6 or 12 J/cm2) and examined the time course of changes in antioxidant enzymes over several days. Only catalase activity was inactivated at the end of the irradiation (66% for 6 J/cm2, 53% for 12 J/cm2), recovering by day 5. Superoxide dismutase (SOD) activity decreased on day 3 (63% for 6 J/cm2, 72% for 12 J/cm2), and recovered on day 5, although it was not changed at the end of exposure. The activities of glutathione peroxidase (GSH-Px) and glutathione reductase (GR) were nearly unchanged by irradiation. If repeated UV exposures occur before the inactivated enzyme activities recover, cellular damage will be significant due to the lowered antioxidant defense system. We examined the rates of synthesis and degradation of catalase in response to UVA irradiation. Both synthesis and degradation rates were changed by irradiation. These data indicate that catalase activity was still low on day 2 due to the existence of inactivated catalase produced at the end of UV irradiation, and catalase activities recovered by day 5 due to a significant increase in the synthesis rate. To elucidate the role of bound NADPH in catalase in response to UV irradiation, we measured the NADPH released from catalase after UVA irradiation using bovine liver catalase. UVA irradiation caused a release of NADPH from catalase (25% for 12 J/cm2), but this was not directly related to the inactivation of catalase.