COCAINE HEPATOTOXICITY - 2 DIFFERENT TOXICITY MECHANISMS FOR PHENOBARBITAL-INDUCED AND NONINDUCED RAT HEPATOCYTES

Hepatocytes isolated from both phenobarbital-induced and control rats were short-term cultured and exposed to cocaine (8-2000 mu M) for varying times. Intracellular lactate dehydrogenase activity, free calcium levels ([Ca2+](i)), reduced glutathione (GSH) and lipid peroxidation were investigated to evaluate the toxic effect of cocaine on hepatocytes. Cytochrome P450 induction by phenobarbital potentiated the in vitro cytotoxicity of cocaine by a factor of 13 (IC50 = 84 mu M in induced cells vs 1100 mu M in non-induced cells). This difference in the susceptibility of the two types of hepatocytes to cocaine correlated well with the activity of cytochrome P450 2B1/2. Rapid depletion of GSH, reaching 30% of the control levels, and massive lipid peroxidation thereafter were the two most remarkable phenomena preceding cell death in phenobarbital-induced hepatocytes. On the other hand, a sustained rise in [Ca2+](i) starting 2 hr after incubation with cocaine was the most noteworthy finding in non-induced liver cells. We suggest two different pathways for cocaine hepatotoxicity: in phenobarbital-induced hepatocytes oxidative metabolism of the drug causes GSH depletion, subsequent extensive lipid peroxidation and cell death, at low concentrations of cocaine. In non-induced hepatocytes these changes are less relevant, and the major alteration caused by cocaine is a non-transient rise in [Ca2+](i) that is evident at higher concentrations of the drug.