Reaction of glutathione with the electrophilic metabolites of 1,1-dichloroethylene.
1,1-Dichloroethylene (DCE) requires cytochrome P450-catalyzed bioactivation to electrophilic metabolites (1,1-dichloroethylene oxide, 2-chloroacetyl chloride and 2,2-dichloroacetaldehyde) to exert its cytotoxic effects. In this investigation, we examined the reactions of these metabolites with glutathione by spectroscopic and chromatographic techniques. In view of the extreme reactivity of 2-chloroacetyl chloride, primary reactions are likely to include alkylation of cytochrome P450, conjugation with GSH to give S-(2-chloroacetyl)-glutathione, or hydrolysis to give 2-chloroacetic acid. Our results showed conjugation of GSH with 1,1-dichloroethylene oxide, through formation of the mono- and di-glutathione adducts, 2-S-glutathionyl acetate and 2-(S-glutathionyl) acetyl glutathione, respectively. The observed equilibrium constant between the hydrate of 2,2-dichloroacetaldehyde and S-(2,2-dichloro-1-hydroxy)ethylglutathione was estimated from 1H-NMR experiments to be 14 +/- 2 M-1. Thus, 2,2-dichloroacetaldehyde is unlikely to make a significant contribution to GSH depletion as GSH concentrations above normal physiological levels would be necessary to form significant amounts of S-(2,2-dichloro-1-hydroxy)ethylglutathione. We also compared the formation of the glutathione conjugates in rat and mouse liver microsomes using 14C-DCE. The results demonstrated a species difference; the total metabolite production was 6-fold higher in microsomes from mice, compared with samples from rat. Production of DCE metabolites in hepatic microsomes from acetone-pretreated mice was 3-fold higher than those from untreated mice suggesting a role for P450 2E1 in DCE bioactivation. These results indicate that the epoxide is the major metabolite of DCE that is responsible for GSH depletion, suggesting that it may be involved in the hepatotoxicity evoked by DCE. Furthermore, this metabolite is formed to a greater extent in mouse than in rat liver microsomes and this difference may underlie the enhanced susceptibility found in the former species.
Dowsley
T F
TF
Department of Anatomy, Queen's University Kingston, Ontario, Canada.
Forkert
P G
PG
Benesch
L A
LA
Bolton
J L
JL
D000079:Acetaldehyde / Q000031:analogs & derivatives / Q000378:metabolism; D000085:Acetates / Q000378:metabolism; D000477:Alkylating Agents / Q000737:chemistry / Q000378:metabolism; D000478:Alkylation; D000818:Animals; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004000:Dichloroethylenes / Q000378:metabolism / Q000633:toxicity*; D005978:Glutathione / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D051379:Mice; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley
D000085:Acetates; D000477:Alkylating Agents; D004000:Dichloroethylenes; C056921:S-(2-chloroacetyl)glutathione; C029297:vinylidene chloride; C006972:chloroacetic acid; D003577:Cytochrome P-450 Enzyme System; C015798:dichloroacetaldehyde; D005978:Glutathione; D000079:Acetaldehyde; C045557:chloroacetyl chloride