Metabolism of 3-methylindole by prostaglandin H synthase in ram seminal vesicles

Abstract

3-Methylindole (3MI) causes a highly tissue- and species-selective lesion of the lung. Metabolic activation of 3MI by the NADPH-dependent mixed function oxidase (MFO) system is the initial event in the lung-specific toxicity. One-electron co-oxidation of 3MI by prostaglandin H synthase (PHS) has been implicated as an alternative mechanism for toxicity in the lung that contains high PHS activity. The objective of this study was to determine if 3MI can be co-oxidized by the arachidonic acid dependent PHS complex. Ram seminal vesicle (RSV) microsomes,which lack MFO activity, were used as a source of PHS. Incubations of RSV microsomes with 3MI, at a concentration as low as 0.01 mM, showed an increase in PHS activity, as indicated by an enhanced rate of oxygen consumption. This effect was arachidonic acid dependent and was inhibited (98%) by indomethacin. Addition of 3MI resulted in a concentration-dependent increase in PHS-catalyzed prostaglandin biosynthesis from [14C]arachidonic acid. PHS-dependent oxidative metabolism of [14C]3MI resulted in a twofold increase in ethyl acetate extracted radiolabelled metabolites. ESR spin-trapping studies demonstrated the presence of a 3MI free radical generated from the metabolism of 3MI by horseradish peroxidase, a model system of PHS hydroperoxidase. The results indicate that 3MI can be co-oxidized by the arachidonic acid-dependent PHS complex. Co-oxidation of 3MI by PHS may play a role in the tissue specificity of 3MI-induced pneumotoxicity.