Formation and disposition of<i>N</i>-hydroxylated metabolites of aniline and nitrobenzene by isolated rat hepatocytes

Abstract

The formation and secretion of phenylhydroxylamine plus nitrosobenzene was studied in cultures of isolated rat hepatocytes after addition of aniline or nitrobenzene. With aniline concn. up to 10 mM, N-oxygenated metabolites were secreted linearly with time over 2 h. Phenobarbitone pretreatment in vivo for c. 5 d increased aniline N-hydroxylation by a factor of 2.8. Nitrobenzene reduction by isolated rat hepatocytes, yielding phenylhydroxylamine plus nitrosobenzene in the medium, was stimulated 1.9-fold and 4.3-fold after phenobarbitone pretreatment in vivo for 5 and 10 d, respectively. After reduction of nitrobenzene by isolated hepatocytes, the secretion of N-oxygenated products into the medium was non-linear with time for substrate concn. higher than 2.5 mM, probably due to the formation of cytotoxic concn. of nitrosobenzene. Isolated rat hepatocytes reduced phenylhydroxylamine to aniline. Results indicate that isolated rat hepatocytes are a reliable and sensitive system to demonstrate N-oxygenated metabolites of aromatic amino- and reduction of nitrocompounds.