Inhibition of the hepatocarcinogenicity of aflatoxin B1 in rats by low levels of the phenolic antioxidants butylated hydroxyanisole and butylated hydroxytoluene.

Abstract

The phenolic antioxidants butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) were studied for inhibition of aflatoxin B1 (AFB1) hepatocarcinogenesis in male Fischer 344 rats. The antioxidants were administered at 5, 25, or 125 ppm in AIN-76A diet for 42 weeks. Beginning with week 2, 5 micrograms/kg of AFB1 was given by intragastric instillation three times a week for 40 weeks either alone or concurrently with BHA or BHT feeding. The development of hepatocellular altered foci (HAF) induced by AFB1, as indicators of hepatocarcinogenesis, was monitored using immunohistochemical staining for the placental form of glutathione S-transferase. By 16 weeks the multiplicity of foci was 1.97/cm2 of liver area in rats given only AFB1, and this increased to 4.11/cm2 at 24 weeks and to 10.60/cm2 at 32 weeks. At the final sacrifice at 42 weeks, the multiplicity of foci was 12.90/cm2 compared to 0.75/cm2 in untreated controls. In rats given antioxidants in addition to AFB1, the high dose of BHA reduced the multiplicity to 7.72/cm2 and the high dose of BHT reduced the multiplicity to 9.35/cm2. Lower levels did not reduce foci induction. Thus, in male rats under the conditions of this experiment, the level of 125 ppm of either BHA or BHT inhibited the initiation of hepatocarcinogenesis by AFB1. The BHA effect was slightly greater than that of BHT.