Dissociation of DDVP-induced DNA strand breaks from oxidative damage in isolated rat hepatocytes.

Dichlorvos (DDVP)-induced DNA single strand breaks were investigated in isolated rat hepatocytes. In a dose-response study in hepatocytes from PB-treated rats (80 mg/kg i.p., for 3 days), 250 microM DDVP substantially reduced cellular non-protein sulfhydryl (NPSH) content, but had no detectable effect on DNA. At 500 microM, the increase in DNA single strand breaks was significant, with a slight increase in cellular lipid peroxidation. At doses over 1000 microM DDVP, cell death was accompanied with considerable lipid peroxidation, and DNA single strand breaks were evident. When the antioxidant N,N'-diphenyl-p-phenylene diamine (DPPD) was added or if the hepatocytes were incubated under air instead of 95% O2, lipid peroxidation and cell death were attenuated but DNA single strand breaks and reduction in NPSH content were not. On the other hand, ferrous iron-induced DNA single strand breaks, lipid peroxidation, and depletion of NPSH content were all attenuated by DPPD or by incubating the cells under air. With respect to the subcellular lipid peroxidation, DDVP caused a significant increase mainly in the microsomal fraction, whereas ferrous iron caused rapid and substantial increases in mitochondrial, microsomal, and nuclear fractions. There were more DNA single strand breaks caused by N-nitrosodiethylamine (NDEA), which becomes genotoxic after microsomal metabolism, in hepatocytes from PB-treated rats than in those from control rats. The number of these breaks was reduced by adding the cytochrome P450 inhibitor metyrapone. On the other hand, the effect of DDVP on DNA was not affected by modification of the cytochrome P450 status. These results suggest that lipid peroxidation induced by DDVP in isolated rat hepatocytes plays a significant role in its cytotoxicity but not in its genotoxicity.