Effect of cytochrome P450 isozyme inhibitors on 3,5‐ dichloroaniline nephrotoxicity in vitro

Abstract

Chlorinated anilines are used in making agricultural chemicals, dyes, industrial compounds, and pharmaceuticals, and can induce nephrotoxicity in vivo and in vitro. 3,5‐Dichloroaniline (3,5‐DCA, 1.0 mM) induced nephrotoxicity in isolated rat renal cortical cells (IRCC) following 90 min exposure. IRCC pretreated with non‐selective cytochrome P450 (CYP) inhibitors partially attenuated 3,5‐DCA toxicity, suggesting that CYPs may play a role in 3,5‐ DCA bioactivation. The purpose of the this study was to further explore the role of CYP mediated 3,5‐DCA bioactivation. IRCC were obtained from male Fischer 344 rats and incubated (4 ×106 cells/ml; 3mL) with shaking (90 min) with dimethyl sulfoxide or 3,5‐DCA (1.0mM). IRCC were pretreated with various CYP inhibitors [isoniazid (1.0 mM), ketoconazole (0.1 mM), omeprazole (0.01 mM), diethyldithiocarbamate (DEDTCA; 0.1 mM), oleandomycin triacetate (0.5 mM), or sulfaphenazole (0.1 mM)] and cytotoxity was determined by measuring lactate dehydrogenase release. Pretreatment with DEDTCA, omeprazole, or sulfaphenazole partially attenuated 3,5‐DCA induced nephrotoxicity, while ketoconazole, isoniazid, oleandomycin triacetate had no effect. These results suggest that 3,5‐DCA is bioactivated via multiple pathways, including the CYP2C family. (Supported by NIH Grant 8P20GM103434 to West Virginia IDeA Network for Biomedical Research Excellence)