Toluene metabolism in isolated rat hepatocytes: effects of in vivo pretreatment with acetone and phenobarbital.
Hepatocytes isolated from control, acetone- and phenobarbital-pretreated rats were used to study the metabolic conversion of toluene to benzyl alcohol, benzaldehyde, benzoic acid and hippuric acid at low (< 100 microM) and high (100-500 microM) toluene concentrations. The baseline formation rates of toluene metabolites (benzyl alcohol, benzoic acid and hippuric acid) were 2.9 +/- 1.7 and 10.0 +/- 2.3 nmol/mg cell protein/60 min at low and high toluene concentrations, respectively. In vivo pretreatment of rats with acetone and phenobarbital increased the formation of metabolites: at low toluene concentrations 3- and 5-fold, respectively; at high toluene concentrations no significant increase (acetone) and 8-fold increase (phenobarbital). Apparent inhibition by ethanol, 7 and 60 mM, was most prominent at low toluene concentrations: 63% and 69%, respectively, in control cells; 84% and 91% in acetone-pretreated cells, and 32% (not significant) and 51% in phenobarbital-pretreated cells. Ethanol also caused accumulation of benzyl alcohol. The apparent inhibition by isoniazid was similar to that of ethanol at low toluene concentrations. Control and acetone-pretreated cells were apparently resistant towards metyrapone; the decrease was 49% and 64% in phenobarbital-pretreated cells at low and high toluene concentrations, respectively. In these cells, the decrease in presence of combined ethanol and metyrapone was 95% (low toluene concentrations). 4-Methyl-pyrazole decreased metabolite formation extensively in all groups. Benzaldehyde was only found in the presence of an aldehyde dehydrogenase inhibitor. Increased ratio benzoic/hippuric acid was observed at high toluene concentrations. These results demonstrate that toluene oxidation may be studied by product formation in isolated hepatocytes. However, the influence of various enzymes in the overall metabolism could not be ascertained due to lack of inhibitor specificity.
Smith-Kielland
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National Institute of Forensic Toxicology, Oslo, Norway.
Ripel
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D000096:Acetone / Q000494:pharmacology*; D000818:Animals; D001547:Benzaldehydes / Q000378:metabolism; D001565:Benzoates / Q000378:metabolism; D019817:Benzoic Acid; D019905:Benzyl Alcohol; D001592:Benzyl Alcohols / Q000378:metabolism; D001667:Binding, Competitive; D002470:Cell Survival / Q000187:drug effects; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005998:Glycine / Q000378:metabolism; D006626:Hippurates / Q000378:metabolism; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D010084:Oxidation-Reduction; D010634:Phenobarbital / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D014050:Toluene / Q000378:metabolism*
D001547:Benzaldehydes; D001565:Benzoates; D001592:Benzyl Alcohols; D065607:Cytochrome P-450 Enzyme Inhibitors; D006626:Hippurates; D007527:Isoenzymes; D000096:Acetone; D014050:Toluene; D019817:Benzoic Acid; D003577:Cytochrome P-450 Enzyme System; D019905:Benzyl Alcohol; C032175:benzaldehyde; D005998:Glycine; D010634:Phenobarbital