Bioactivation of 1,1-Dichloroethylene by CYP2E1 and CYP2F2 in Murine Lung

• Published in: The Journal of pharmacology and experimental therapeutics 2004-09, Vol.310 (3), p.855-864
• Main Authors: Simmonds, Andrea C, Ghanayem, Burhan I, Sharma, Ashish, Reilly, Christopher A, Millen, Brandie, Yost, Garold S, Forkert, Poh-Gek
• Format: Article
• Language: English
• Subjects: Animals; Blotting, Western; Chlorzoxazone - metabolism; Cytochrome P-450 CYP2E1 - metabolism; Cytochrome P-450 Enzyme System - metabolism; Cytosol - metabolism; Dichloroethylenes - metabolism; Female; Hydroxylation; Lung - enzymology; Lung - metabolism; Mice; Nitrophenols - metabolism
• ISSN: 0022-3565; 1521-0103
• DOI: 10.1124/jpet.104.068171

1,1-Dichloroethylene (DCE) exposure evokes lung toxicity with selective damage to bronchiolar Clara cells. Recent in vitro studies have implicated CYP2E1 and CYP2F2 in the bioactivation of DCE to 2- S -glutathionyl acetate [C], a putative conjugate of DCE epoxide with glutathione. An objective of this study was to test the hypothesis that bioactivation of DCE is catalyzed by both CYP2E1 and CYP2F2 in murine lung. Western blot analysis of lung microsomal proteins from DCE-treated CD-1 mice showed time-dependent loss of immunodetectable CYP2F2 and CYP2E1 protein. Dose-dependent formation of conjugate [C] was observed in the lungs of CD-1 mice treated with DCE (75–225 mg/kg), but it was not detected after pretreatment with 5-phenyl-1-pentyne (5-PIP). Treatment of mice with 5-PIP and also with diallyl sulfone (DASO 2 ) significantly inhibited hydroxylation of p- nitrophenol (PNP) and chlorzoxazone (CHZX). Incubation of recombinant CYP2F3 (a surrogate for CYP2F2) and recombinant CYP2E1 with PNP and CHZX confirmed that they are substrates for both of the recombinant enzymes. Incubation of the recombinant enzymes with DASO 2 or 5-PIP significantly inhibited hydroxylation of both PNP and CHZX. Bronchiolar injury was elicited in CD-1 mice treated with DCE (75 mg/kg), but it was abrogated with 5-PIP pretreatment. Bronchiolar toxicity also was manifested in the lungs of CYP2E1-null and wild-type mice treated with DCE (75 mg/kg), but protection ensued after pretreatment with 5-PIP or DASO 2 . These results showed that bioactivation of DCE in murine lung occurred via the catalytic activities of both CYP2E1 and CYP2F2 and that bioactivation by these enzymes mediated the lung toxicity.