EFFECTS OF MODULATION OF HEPATIC GLUTATHIONE ON BIOTRANSFORMATION AND COVALENT BINDING OF AFLATOXIN-B1 TO DNA IN THE MOUSE

Relative to the rat and most other species tested, the mouse is resistant to the carcinogenic effects of aflatoxin B1 (AFB). Previous investigations in our laboratory demonstrated that mouse liver cytosol has 52 times greater hepatic glutathione S-transferase (GST) activity toward the AFB-epoxide, compared with rat liver cytosol. To determine the importance of GST-mediated detoxification of the AFB-epoxide in the mouse in vivo, we examined the effects of glutathione (GSH) depletion on the covalent binding of AFB to hepatic DNA in control and 2(3)-butyl-4-hydroxyanisole (BHA)-treated mice. Male Swiss-Webster mice were fed control or 0.75% BHA diet for 14 days. Depletion of hepatic GSH was accomplished with D,L-buthionine-S-sulfoximine (BSO, 0.6 g/kg in saline) and diethyl maleate (DEM, 0.75 ml/kg), administered by ip injection at 2 and 1.5 hr, respectively, prior to administration of 3H-AFB (0.25 mg/kg, ip in DMSO). The combined BSO-DEM treatment depleted hepatic GSH by 97 and 70% in control and BHA-treated mice, respectively. In mice receiving the control diet, GSH depletion was associated with a 30-fold increased in the covalent binding of AFB to hepatic DNA. AFB-DNA binding in mice treated with dietary BHA alone was reduced to 54% of control. In BHA-treated mice, BSO-DEM treatment increased AFB-DNA binding by 62%. Dietary BHA increased hepatic S-9 mediated activation of AFB to the AFB-epoxide by eightfold in both control and BSO-DEM mice. BHA also increased GST activity toward the AFB-epoxide by 52 and 68% in control and BSO-DEM mice, respectively. The BSO-DEM treatment alone had no significant effect on the in vitro biotransformation of AFB. These results support the hypothesis that GST is the principal determinant of species differences in susceptibility to AFB-induced hepatocarcinogenicity. The results also support the hypothesis that BHA may protect against the toxic and carcinogenic effects of xenobiotics in part by preventing the depletion of hepatic GSH.