Arginine increases genotoxicity induced by methyl methanesulfonate in human lymphocytes

Abstract

Nitric oxide (NO) is a free radical that is produced in cells from L-arginine. NO is involved in the physiological control of different tissues, but it can act as a toxic mediator in the cells. In this study we investigated the effect of L-arginine on the genotoxicity induced by methyl methanesulfonate (MMS) in human lymphocytes. Blood was treated with N(G)-nitro-L-arginine methyl ester (L-NAME) as an inhibitor of nitric oxide synthase for finding out the role of NO in this effect. Human whole blood was treated with L-arginine (50, 100 and 250μM) and/or L-NAME, then it was treated in vitro with MMS after 24h of culture. The lymphocytes were stimulated by phytohemagglutinin to find out the micronuclei in cytokinesis-blocked binucleated cells. DNA fragmentation of lymphocytes was detected by using a fluorescence microscope after propidium iodide staining. These data showed that arginine increased the frequency of MMS-induced micronuclei in lymphocytes. However, the genotoxicity was decreased by using L-NAME. Arginine and L-NAME have not shown any DNA damage in cultured human lymphocytes. In conclusion, addition of L-arginine to MMS as an alkylating agent caused an increase of DNA damage in human lymphocytes. This enhancement of genotoxicity was reduced by NAME as NO inhibitor. It is thus cleared that an increase of DNA damage by arginine and MMS is related to NO production.