Received for publication July 29, 2005.
Revised November 1, 2005.
Accepted for publication November 2, 2005.

Pulmonary Bronchiolar Cytotoxicity and Formation of Dichloroacetyl Lysine Protein Adducts in Mice Treated with Trichloroethylene

Abstract

This study was undertaken to test the hypothesis that bronchiolar damage induced by trichloroethylene (TCE) is associated with bioactivation within the Clara cells with involvement of CYP2E1 and CYP2F2. Histopathology confirmed dose-dependent Clara cell injury and disintegration of the bronchiolar epithelium in CD-1 mice treated with TCE doses of 500 to 1000 mg/kg, i.p. Immunohistochemical studies, using an antibody that recognizes dichloroacetyl lysine adducts, revealed dose- dependent formation of adducts in the bronchiolar epithelium. Localization of dichloroacetyl adducts in the Clara cells coincided with damage to this cell type in TCE-treated mice. Pretreatment of CD-1 mice with diallyl sulfone, an inhibitor of CYP2E1 and CYP2F2, abrogated formation of the dichloroacetyl adducts and protected against TCE-induced bronchiolar cytotoxicity. Treatment of wild-type and CYP2E1-null mice with TCE (750 mg/kg, i.p.) also elicited bronchiolar damage that correlated with formation of adducts in the Clara cells. Immunoblotting, using lung microsomes from TCE-treated CD- 1 mice, showed dose-dependent production of dichloroacetyl adducts that co-migrated with CYP2E1 and CYP2F2. However, TCE treatment resulted in loss of immunoreactive CYP2E1 and CYP2F2 proteins and p- nitrophenol hydroxylation, a catalytic activity associated with both P450 enzymes. The TCE metabolite, chloral hydrate, was formed in incubations of TCE with lung microsomes from CD-1, wild-type and CYP2E1-null mice. The levels were higher in CD-1 than in either wild- type or CYP2E1-null mice, although levels were higher in CYP2E1-null than in wild-type mice. These findings supported the contention that TCE bioactivation within the Clara cells, predominantly involving CYP2F2, correlated with bronchiolar cytotoxicity in mice.

Key words: CYP2E1, CYP2F2, Clara cells, chloral hydrate, diallyl sulfone, protein adducts

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