PubMed1981
The metabolism of [1,2-14C]vinyl bromide (VBR) to products irreversibly bound to DNA and protein was examined in rat liver microsomes, reconstituted cytochrome P-450 systems, and isolated hepatocytes. A role for cytochrome P-450 was confirmed using inhibition and reconstitution experiments. The major form of cytochrome P-450 involved in VBR metabolism does not appear to be either of the major isozymes induced by phenobarbital or beta-naphthoflavone, as determined by induction, reconstitution, and antibody inhibition studies. 2-Bromoethylene oxide and 2-bromoacetaldehyde, suspected metabolites of VBR, were synthesized and found to be substrates for rat liver epoxide hydrolase and equine liver alcohol dehydrogenase, respectively. These enzymes were used to probe the roles of the two possible metabolites in the irreversible binding of products of VBR to protein and DNA. Alcohol dehydrogenase was more effective than epoxide hydrolase in inhibiting the binding of VBR metabolites to protein in microsomal incubations. Epoxide hydrolase was effective in inhibiting the binding of VBR or vinyl chloride metabolites to calf thymus DNA added to such systems, but alcohol dehydrogenase was not. Similar results were obtained for binding of VBR metabolites to DNA in a reconstituted enzyme system. Reduced glutathione blocked nonenzymatic binding of 2-bromo[1,2-14C]acetaldehyde to protein but not DNA. Binding of vinyl chloride and VBR metabolites to protein was blocked by reduced glutathione, but binding to DNA was not. These results are consistent with the view that 2-haloethylene oxides are the major alkylating agents bound to DNA, and 2-haloacetaldehydes are the major alkylating agents bound to protein in these experimental systems. Studies with labeled 2-bromoacetaldehyde indicate that the slow kinetics of DNA binding by this compound is responsible in part for this phenomenon. Studies with isolated rat hepatocytes suggest that a significant portion of the total and reactive metabolites are able to leave these cells. In these systems, binding of metabolites of vinyl chloride to DNA outside the hepatocytes could be partially blocked by epoxide hydrolase or by alcohol dehydrogenase, implying that, as target farther away from sources of reactive species are considered, the stabilities of these species become more important for reaction with nucleophilic sites.