Time course of 72-kilodalton heat shock protein induction and appearance of trifluoroacetyl adducts in livers of halothane-exposed rats

WQ Lin, RA van Dyke, HM Marsh and JR Trudell

Department of Anesthesia, Standford School of Medicine, California 94305-5117.

Previous studies have shown that exposure of phenobarbital-pretreated rats to halothane in 10% O2 causes centrilobular necrosis, induces expression of the 72-kDa heat shock protein (HSP72), and produces several trifluoroacetylated adducts. In the present study the time course of development of the centrilobular lesion, as measured by histochemistry, was compared with the time course of appearance of both trifluoroacetylated adducts and HSP72, as measured by Western blotting. One group of 20 rats was pretreated with phenobarbital for 5 days, whereas a second group of two rats was left as untreated controls. Ten phenobarbital-pretreated rats were exposed for 2 hr to 1% halothane in 10% O2 and 10 were exposed to 1% halothane in 20% O2. At either 2, 4, 6, or 24 hr after exposure, livers were excised and frozen without fixation. Thin sections stained with hematoxylin and eosin demonstrated that centrilobular lesions occurred at 6 hr and became extensive at 24 hr in rats pretreated with phenobarbital and exposed to 1% halothane in 10% O2. The time course of appearance of both trifluoroacetylated adducts and HSP72 was determined by Western blotting. Trifluoroacetylated adducts appeared in all rats exposed to halothane by 2 hr, lasted until 6 hr, and then diminished by 24 hr. In contrast, HSP72 was induced only in the rats pretreated with phenobarbital and exposed to 1% halothane in 10% O2. HSP72 appeared in both the nuclear and supernatant fractions at 6 hr after exposure and was intense 24 hr after exposure.

Volume 46, Issue 4, pp. 639-643, 10/01/1994
Copyright © 1994 by American Society for Pharmacology and Experimental Therapeutics