Effect of hydrogen peroxide on nitric oxide (NO)-induced mutagenicity in salmonella typhimurium

Abstract

Nitric oxide (NO) has been reported to impart, alone or in combination with reactive oxygen species (ROS), the cytotoxicity and putative genotoxicity associated with the immunological response. The present study examined the change in the mutagenic activity profile of the NO-donor spermine NONOate (SperNO) as a result of introduction of hydrogen peroxide (H2O2) to the Ames assay. The aim was to determine whether the assay could detect H2O2-induced co- or anti-mutagenic effects on NO-induced mutagenesis, and the Salmonella typhimurium base-pair substitution tester strain TA1535 provided an appropriate tool. While TA1535 was shown by the authors and others to be strongly sensitive to NO-induced mutagenesis, it has also been shown to be insensitive to H2O2-induced mutagenicity [1,2]. When H2O2 (0.25-4.0 μmol/pl) was added directly to cells treated with SperNO (0.01-1.0 μmol/pl), comutagenicity was not detected, but a drop in reversion count and detectable toxicity was observed, especially at doses > 0.1 μmol/pl. When glucose/glucose oxidase (GOX) or reduced glutathione (GSH) were used as H2O2-generation systems the results varied. Reversion induced by SperNO (1 μmol/pl) was moderately enhanced by GOX (10-20 mUnits/pl), but the increase albeit reproducible did not reach a doubling (co-mutagenicity). GOX (40 μmol/pl) induced a reduction in reversion count, but no visible toxicity. On the other hand, GSH (20- 80 μmol/pl) gave a strong co-mutagenic effect. Co-mutagenicity was highest (> 5x) at 80 μmol/pl GSH and 0.1 μmol/pl SperNO. Based on these findings, it could be concluded that a) H2O2, when steadily generated in the cell, has a modulatory effect on NO-mutagenicity, and such a conclusion is not inconsistent with the wide range of responses reported for the two chemicals, and/or b) the observed co-mutagenic effects of GSH may not be attributable solely to H2O2 generation. © 2001 Wiley-Liss, Inc.