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Effects of 2- and 3-tert-Butyl-4-hydroxyanisole on Glutathione S-Transferase and Epoxide Hydrolase Activities and Sulfhydryl Levels in Liver and Forestomach of Mice¹

Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, Minnesota 55455

Butylated hydroxyanisole, a food additive, has been found to inhibit the neoplastic effects of a wide variety of chemical carcinogens. The commercially available preparations of butylated hydroxyanisole contain two isomers, 2-tert-butyl-4-hydroxyanisole (2-BHA) and 3-tert-butyl-4-hydroxyanisole (3-BHA). Both isomers induce increased activities of glutathione (GSH) S-transferase and epoxide hydrolase and increase acidsoluble sulfhydryl concentration in hepatic and forestomach tissues of A/HeJ mice. The inductions were assayed after 2 weeks of feeding diets containing the two isomers. 3-BHA induced an increase in the activity of hepatic microsomal epoxide hydrolase by 1.4 times that of the control. The activity of cytosolic GSH S-transferase was enhanced by both isomers. In the liver, the 3-BHA induction was more than 3 times higher than that of 2-BHA. In the forestomach, however, the induction effect of the two isomers was reversed. The overall magnitude of the induction was much lower in the forestomach than in the liver. Synergistic effects on the induction of GSH S-transferase activity were observed in the forestomach cytosol when mixtures of different proportions of the two isomers were added to the diet. Maximum enzyme activity was obtained at a ratio of 75% 2-BHA and 25% 3-BHA. No synergistic effect was observed with the corresponding hepatic cytosolic enzyme. The relative inductive effects of 2- and 3-BHA on the acid-soluble sulfhydryl level of liver and forestomach tissues followed closely those on GSH S-transferase activity. The results of the present study show that the two isomers of butylated hydroxyanisole differ in the magnitude of their effects on carcinogenmetabolizing systems of the liver and forestomach.

¹ Supported by USPHS Grants CA-14146 and CA-09599 from the National Cancer Institute. Presented in part at the Second Chemical Congress of the North American Continent, Las Vegas, Nev., August 24 to 29, 1980 (14).

² To whom requests for reprints should be addressed.

Received 2/17/81. Accepted 7/10/81.

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