Carbendazim and n-butylisocyanate: metabolites responsible for benomyl double action on cytochrome P450 in HepG2 cells.

Changes in the cytochrome P450 monooxygenase system were investigated in HepG2 cells treated for 24 h with 1.25, 2.5, 5, 10 and 20 microg/ml of carbendazim (MBC) and n-butylisocyanate (BIC), the principal benomyl metabolites. The results show that n-butylisocyanate leads to a decrease in both ethoxyresorufin deethylase (P4501A1) (EROD) and ethoxycoumarin deethylase (P4502B) (ECOD), whereas MBC has no effect on EROD and increases ECOD. The decrease in ECOD and EROD activities after BIC treatment can be attributed to the detrimental action of this substance. The MBC-induced increase in ethoxycoumarin can be considered an enzyme-specific inductive phenomenon. This hypothesis was confirmed by Western immunoblot analysis and treatment with actinomycin D 8 x 10(-4) microM: the first showed an increase in P4502B isoenzyme content and the second evidence of a partial block of the increase in ECOD activity induced by MBC. Given these results, MBC and BIC seem to be the metabolites responsible for the double opposite action of their parent compound benomyl. Data deriving from an equimolar mixture of the two metabolites suggest that benomyl activity on some cytochrome P450 isoenzymes is the result of a balance between the action of the single metabolites (Radice et al., 1996).