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4‐Amino‐2‐chlorophenol nephrotoxicity in vitro: alteration of cytotoxicity by antioxidants
Author(s) -
Rankin Gary OˈNeal,
Ferguson Travis,
Baksi Sukanya,
Anestis Dianne,
Racine Christopher
Publication year - 2012
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.26.1_supplement.1051.10
Subject(s) - chemistry , glutathione , nephrotoxicity , cytotoxicity , pharmacology , antioxidant , metabolite , biochemistry , oxidative stress , toxicity , enzyme , in vitro , biology , organic chemistry
4‐Amino‐2‐chlorophenol (4A2CP, 1.0 mM) induces nephrotoxicity in isolated rat renal cortical cells (IRCC). The purpose of this study was to examine potential metabolic pathways (cytochrome P450 [CYP], flavin mono‐oxygenase [FMO], and oxidation‐reduction) for bioactivation of 4A2CP. IRCC were obtained from male Fischer 344 rats and incubated in Krebs‐Henseleit buffer. IRCC (4 million cells/ml; 3 ml) were incubated with dimethyl sulfoxide (DMSO) or 4A2CP (1.0 mM) for 60 min with shaking. In some experiments, IRCC were pretreated with an FMO inhibitor [N‐octylamine (0.2 mM) or methimazole (1.0 mM)], CYP inhibitor [isoniazid (1.0 mM), ketoconazole (1.0 mM), or metyrapone (1.0 mM)], or an antioxidant [ascorbate (1.0 mM), glutathione (1.0 mM), N‐acetyl‐L‐cysteine (NAC, 2.0 mM), DPPD (0.05 mM), deferoximine (0.1 mM), or alpha‐tocopherol (1.0 mM or 2.0 mM)]. Cytotoxicity was assessed by measuring the release of lactate dehydrogenase (LDH). FMO or CYP inhibitors didn't decrease 4A2CP cytotoxicity. Only ascorbate, glutathione and NAC provided protection from 4A2CP toxicity. These results suggest that the mechanism by which 4A2CP induces toxicity does not involve activation by FMO's, CYP2E1, 2B or 3A4. Antioxidant protection suggests that a toxic metabolite is created via an auto‐oxidation process and/or oxidative stress plays a role in 4A2CP nephrotoxicity. (Supported by NIH Grant 5P20RR016477 to WV‐INBRE)