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Nephrotoxic Potential of 3,5‐Dichloroaniline Putative Metabolites in Isolated Renal Cortical Cells from Fischer 344 Rats
Author(s) -
Rankin Gary,
Racine Christopher,
Ward Dakota,
Tyree Conner,
Pope Deborah,
Anestis Dianne
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.711.3
Subject(s) - metabolite , nephrotoxicity , chemistry , biotransformation , cytotoxicity , in vivo , lactate dehydrogenase , biochemistry , pharmacology , kidney , enzyme , toxicity , in vitro , biology , endocrinology , organic chemistry , microbiology and biotechnology
Halogenated anilines are widely used as chemical intermediates in the manufacture of dyes, agricultural agents and as industrial intermediates. Previous studies in our laboratory have found that 3,5‐dichloroaniline (3,5‐DCA) is the most potent nephrotoxicant among mono‐ and dichloroanilines, both in vivo and in vitro. The potential role of renal biotransformation in 3,5‐DCA‐induced nephrotoxicity has also been examined. When isolated renal cortical cells (IRCC) from male Fischer 344 rats were pretreated with various biotransformation enzyme inhibitors, 3,5‐DCA –induced cytotoxicity was significantly attenuated. These studies indicated that 3,5‐DCA could be biotransformed via multiple pathways in the kidney to one or more toxic metabolite(s). The current study was designed to explore the nephrotoxic potential of three putative 3,5‐DCA metabolites [2‐amino‐4,6‐dichlorophenol (2A4,6DCP), 3,5‐dichlorophenylhydroxylamine (3,5‐DCPHA), and 3,5‐dichloroacetanilide (3,5‐DCAA)] in IRCC from male Fischer 344 rats. Briefly, IRCC were isolated using a collagenase perfusion technique, and IRCC (~4.1 million cells/ml; 3 ml) were exposed to various concentrations of a putative metabolite [2A4,6DCP or 3,5‐DCAA (0.5–1.5 mM); 3,5‐DCPHA (0.25–1.0 mM)]or vehicle (DMSO) for 60 or 90 min. Cytotoxicity was determined by lactate dehydrogenase (LDH) release assays. Results showed that there were no significant changes in LDH release following exposure to 3,5‐DCAA at any time point or concentration tested. However, 2A4,6DCP was a nephrotoxicant at all concentrations tested, but only after 90 min exposure. In contrast, 3,5‐DCPHA significantly increased LDH release at concentrations of 0.5 mM or greater after 60 min and at all concentrations tested after exposure for 90 min. These results suggest that at least two putative 3,5‐DCA metabolites [3,5‐DCPHA and/or 2A4,6DCP, but not 3,5‐DCAA] may contribute to 3,5‐DCA induced nephrotoxicity in vitro. Support or Funding Information Supported in part by NIH grant P20GM103434