Interaction of trichloroethane isomers with cytochrome P-450 in the perfused rat liver

Abstract

The real-time interactions of 1,1,1-trichloroethane (TCE) and 1,1,2-TCE with cytochrome P-450 were observed using in vivo optical methods to measure the spectral changes of cytochrome P-450 and the reduction-oxidation transition of pyridine nucleotides in the perfused liver of rats treated with phenobarbital. Changes in oxygen consumption and TCE uptake were also measured. The spectral changes of cytochrome P-450 indicated that both TCE isomers bound to low spin (substrate free) ferric cytochrome P-450 and formed a high spin (substrate complexed) form. However, 1,1,1-TCE bound more tightly to cytochrome P-450 and seemed to be only slowly metabolized compared to 1,1,2-TCE. The stoichiometry of the change in oxygen consumption rate to the change in 1,1,1-TCE uptake rate ranged between 5 1 and 9 1 , whereas that of 1,1,2-TCE was 1.4 to 2.0. Decreases in reduced pyridine nucleotides associated with TCE administration were significantly larger with 1,1,1-TCE than with 1,1,2-TCE. The inhibitory effect of 1,1,1-TCE on hexobarbital metabolism in the perfused liver was greater than that of 1,1,2-TCE. Considering our previous data indicating that TCE did not stimulate mitochondrial respiration, it is postulated that the far higher amount of oxygen consumption associated with the binding of 1,1,1-TCE to cytochrome P-450 than the amount which was necessary to mixed-function oxidation of this compound was due to an uncoupling effect of 1,1,1-TCE on the mixed-function oxidase system.