title: Chlorpropham induces mitochondrial dysfunction in rat hepatocytes.
abstract: The metabolism and action of chlorpropham (isopropyl N-(3-chlorophenyl)carbamate; CIPC, a post-harvest agent) and its metabolites were studied in freshly isolated rat hepatocytes and isolated rat hepatic mitochondria, respectively. The exposure of hepatocytes to CIPC caused a concentration (0.25-1.0 mM)- and time (0-3h)-dependent cell death accompanied by loss of cellular ATP and adenine nucleotides. CIPC at a weakly toxic level (0.5 mM) was metabolized to isopropyl N-(3-chloro-4-hydroxyphenyl)carbamate (4OH-CIPC) and subsequently to its glucuronide and sulfate conjugates (major metabolites) or alternatively to a minor metabolite 3-chloroaniline (3CA). The addition of SKF-525A (50 microM), an inhibitor of microsomal monooxygenase, enhanced the CIPC (0.5 mM)-induced cytotoxicity accompanied by loss of ATP and 4OH-CIPC and inhibited the decrease in the concentration of the parent compound. CIPC led to a strong decrease in cellular ATP content compared to its metabolites, 4OH-CIPC and 3CA. On the other hand, the exposure of isolated hepatic mitochondria to CIPC reduced State 3 respiration with a FAD-linked substrate (succinate plus rotenone) and/or with a NAD+ -linked substrate (pyruvate plus malate), whereas State 3 respiration with ascorbate plus tetramethyl-p-phenylendiamine (cytochrome oxidase-linked respiration) was not affected markedly by CIPC. Further, the addition of CIPC caused an increase in the rate of State 4 oxygen consumption, indicating an uncoupling effect, and a decrease in the rate of State 3 oxygen consumption in a concentration-dependent manner, respectively. In contrast, the addition of neither 4OH-CIPC nor 3CA markedly affected the rate of states 3 and/or 4 oxygen consumption. These results indicate that CIPC-induced cytotoxicity is mediated by the parent compound rather than by its metabolites such as 4OH-CIPC and 3CA, and that the toxicity is associated with a rapid depletion of ATP via impairment of mitochondrial function related to oxidative phosphorylation.
authors.affiliation: Divisions of Pharmacology, Tokyo Metropolitan Institute of Public Health, 3-24-1 Hyakunin-cho, Shinjuku-ku, 169-0073, Japan. yoshio_1_nakagawa@member.metro.tokyo.jp
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Nakagawa
authors.forename: Kazuo
authors.initials: K
authors.lastname: Nakajima
authors.forename: Toshinari
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D000227:Adenine Nucleotides / Q000378:metabolism; D000818:Animals; D001711:Biotransformation; D016923:Cell Death / Q000187:drug effects; D002469:Cell Separation; D002748:Chlorpropham / Q000378:metabolism / Q000633:toxicity*; D002851:Chromatography, High Pressure Liquid; D004791:Enzyme Inhibitors / Q000633:toxicity; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism*; D006540:Herbicides / Q000378:metabolism / Q000633:toxicity*; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D013058:Mass Spectrometry; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors; D010101:Oxygen Consumption / Q000187:drug effects; D011335:Proadifen / Q000494:pharmacology; D051381:Rats; D011916:Rats, Inbred F344
chemical_list: D000227:Adenine Nucleotides; D004791:Enzyme Inhibitors; D006540:Herbicides; D002748:Chlorpropham; D011335:Proadifen; D006899:Mixed Function Oxygenases