In vitro studies on the metabolism and covalent binding of [14C]1,1-dichloroethylene by mouse liver, kidney and lung.
The metabolism and covalent binding of 1,1-dichloro[1,2-14C]ethylene (DCE) to subcellular fractions of liver, kidney and lung of C57BL/6N mice have been investigated in vitro. Covalent binding was NADPH- and cytochrome P-450-dependent. The microsomal fraction bound more radiolabel than any other subcellular fraction, and the levels of covalent binding in cell fractions correlated well with their cytochrome P-450 content. Covalent binding by mouse liver and lung microsomes also reflected their cytochrome P-450 content. However, although mouse kidney microsomes contained twice as much total cytochrome P-450 as the lung, no detectable covalent binding of DCE-derived radioactivity occurred in kidney. Omission of NADPH, heat inactivation of microsomes, carbon monoxide, addition of SKF-525A, piperonyl butoxide or reduced glutathione (GSH), all inhibited (40-90%) covalent binding of radiolabel to liver and lung microsomes. The absence of O2 (incubation under N2) did not greatly affect the metabolism and covalent binding. Pretreatment of mice with various inducers, phenobarbital (PB), beta-naphthoflavone (beta-NF), pregnenolone 16 alpha-carbonitrile (PCN) and 3-methylcholanthrene (3-MC), evoked increases in total liver microsomal cytochrome P-450 content (2-fold) and corresponding increases in covalent binding (3-fold). However, microsomes from PCN-treated mice showed only a 50% increase in DCE binding. Kidney microsomes from control, PB-, and beta-NF-pretreated mice were incapable of covalent binding of radiolabel but those from PCN- and 3-MC-pretreated mice showed levels of binding similar to untreated mouse lung microsomes. It is proposed that the nephrotoxicity of DCE may be due to translocation of reactive metabolites from the liver to the kidney.
D000818:Animals; D001571:Benzoflavones / Q000494:pharmacology; D002248:Carbon Monoxide / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism; D004000:Dichloroethylenes / Q000378:metabolism*; D004790:Enzyme Induction; D005978:Glutathione / Q000494:pharmacology; D006358:Hot Temperature; D006843:Hydrocarbons, Chlorinated / Q000378:metabolism*; D066298:In Vitro Techniques; D007668:Kidney / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008168:Lung / Q000378:metabolism*; D008748:Methylcholanthrene / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D009249:NADP / Q000378:metabolism; D010634:Phenobarbital / Q000494:pharmacology; D010882:Piperonyl Butoxide / Q000494:pharmacology; D011285:Pregnenolone Carbonitrile / Q000494:pharmacology; D011335:Proadifen / Q000494:pharmacology; D013347:Subcellular Fractions / Q000378:metabolism; D019324:beta-Naphthoflavone
D001571:Benzoflavones; D004000:Dichloroethylenes; D006843:Hydrocarbons, Chlorinated; D011285:Pregnenolone Carbonitrile; C029297:vinylidene chloride; D009249:NADP; D008748:Methylcholanthrene; D019324:beta-Naphthoflavone; D002248:Carbon Monoxide; D003577:Cytochrome P-450 Enzyme System; D011335:Proadifen; D005978:Glutathione; D010882:Piperonyl Butoxide; D010634:Phenobarbital