Drug-induced hydrogen peroxide production in isolated rat hepatocytes

A method for measuring drug-induced hydrogen peroxide production in freshly isolated rat hepatocytes is described. 3-Amino-1,2,4-triazole, an irreversible inhibitor of the enzyme catalase markedly reduces the capacity of isolated hepatocytes to metabolize hydrogen peroxide, with maximum inhibition (80%) being observed after 40 min of co-incubation. The present method is based on the observation that this inhibition of catalase by 3-amino-1,2,4-triazole is prevented by methanol and that the effect of methanol is reversed in the presence of hydrogen peroxide. Using this assay we could demonstrate increased hydrogen peroxide production during the metabolism of diquat, paraquat, xanthine, benzylamine and glycolate by hepatocytes. Inhibition of the hydrogen peroxide metabolic capacity was greatest with glycolate and diquat, whereas paraquat and benzylamine only had a minor effect.