The site specificity and sensitivity of the rat liver to butylated hydroxytoluene-induced damage

The food additive butylated hydroxytoluene (BHT) is capable of damaging centrilobular or periportal cells in the liver according to the dose and duration of treatment. The effect of two hepatotoxicity potentiating agents on the site specificity of acute cell damage was investigated in Sprague-Dawley rats. A 500 mg/kg oral dose of BHT did not cause overt hepatic necrosis or alter the cytochrome P450 concentration, but increased ethoxycoumarin-O-deethylation, implying an alteration in the ratio of P450 isoenzymes. Pretreatment with either phenobarbitone (3 X 80 mg/kg, ip) or the glutathione depleting agent buthionine sulfoximine (900 mg/kg, ip) produced liver necrosis in approximately 50% of animals: mainly in centrilobular areas, but with some necrosis in midzonal or periportal areas. Phenobarbitone and BHT did not significantly change the cytochrome P450 concentration, but did alter the ratio of P450 isoenzymes. In phenobarbitone-pretreated rats centrilobular hepatocyte damage was clearly localized in cells with high immunocytochemical staining for the cytochrome P450IIB subfamily. Buthionine sulfoximine and BHT reduced the cytochrome P450 concentration without reducing ethoxycoumarin-O-deethylase activity, implying a different alteration in the ratio of P450 isoenzymes. These results indicate that phenobarbitone-inducible enzymes are capable of activating high doses of BHT to reactive oxidizing intermediates, which in the absence of adequate glutathione can cause cell death. Enzymes of the P450IIB subfamily are implicated in this mechanism.