The role of biotransformation and oxidative stress in 3, 5-dichloroaniline (3, 5-DCA) induced nephrotoxicity in isolated renal cortical cells from male Fischer 344 rats. (3rd February 2016)

• Record Type: Journal Article
• Authors: Racine, Christopher R.; Ferguson, Travis; Preston, Debbie; Ward, Dakota; Ball, John; Anestis, Dianne; Valentovic, Monica; Rankin, Gary O.
• Is Part Of: Toxicology. Volume 341/343(2016)
• Page Start: 47
• Page End: 55
• Publication Date: 2016-02-03
• Subjects: 3, 5-DCA 3, 5-dichloroaniline -- IRCC isolated renal cortical cells -- LDH lactate dehydrogenase -- ALT alanine aminotransferase -- GPT glutamic-pyruvic transaminase -- GSH glutathione -- GSSG oxidized glutathione -- DMSO dimethyl sulfoxide -- DNP 2, 4-dinitrophenylhydrazone -- FMO flavin-containing monooxygenase -- CYP cytochrome P450 -- PiBx piperonyl butoxide -- DEDTCA diethyldithiocarbamate -- ASC ascorbate -- NAC N-acetyl-l-cysteine -- 3, 5-DCAA 3, 5-dichloroacetanilide -- 3, 5-DCPHA 3, 5-dichlorophenylhydroxylamine -- 3, 5-DCNB 3, 5-dichloronitrobenzene; Nephrotoxicity -- In vitro -- Rat -- 3, 5-Dichloroaniline -- Oxidative stress; Toxicology -- Periodicals; Chemicals -- Physiological effect -- Periodicals; 615.9005
• DOI: 10.1016/j.tox.2016.01.006
• Languages: English
• ISSNs: 0300-483X

Abstract

Abstract: Among the mono- and dichloroanilines, 3, 5-dichloroaniline (3, 5-DCA) is the most potent nephrotoxicant in vivo and in vitro. However, the role of renal biotransformation in 3, 5-DCA induced nephrotoxicity is unknown. The current study was designed to determine the in vitro nephrotoxic potential of 3, 5-DCA in isolated renal cortical cells (IRCC) obtained from male Fischer 344 rats, and the role of renal bioactivation and oxidative stress in 3, 5-DCA nephrotoxicity. IRCC (∼4 million cells/ml) from male rats were exposed to 3, 5-DCA (0–1.0 mM) for up to 120 min. In IRCC, 3, 5-DCA was cytotoxic at 1.0 mM by 60 min as evidenced by the increased release of lactate dehydrogenase (LDH), but 120 min was required for 3, 5-DCA 0.5 mM to increase LDH release. In subsequent studies, IRCC were exposed to a pretreatment (antioxidant or enzyme inhibitor) prior to exposure to 3, 5-DCA (1.0 mM) for 90 min. Cytotoxicity induced by 3, 5-DCA was attenuated by pretreatment with inhibitors of flavin-containing monooxygenase (FMO; methimazole, N -octylamine), cytochrome P450 (CYP; piperonyl butoxide, metyrapone), or peroxidase (indomethacin, mercaptosuccinate) enzymes. Use of more selective CYP inhibitors suggested that the CYP 2C family contributed to 3, 5-DCA bioactivation. Antioxidants (glutathione, N -acetyl-l -cysteine, α-tocopherol, ascorbate, pyruvate) also attenuated 3, 5-DCA nephrotoxicity, but oxidized glutathione levels and the oxidized/reduced glutathione ratios were not increased. These results indicate that 3, 5-DCA may be activated via several renal enzyme systems to toxic metabolites, and that free radicals, but not oxidative stress, contribute to 3, 5-DCA induced nephrotoxicity in vitro.