Ramdhan Doni Hikmat, Kamijima Michihiro, Yamada Naoyasu, Ito Yuki, Yanagiba Yukie, Nakamura Daichi, Okamura Ai, Ichihara Gaku, Aoyama Toshifumi, Gonzalez Frank J, Nakajima Tamie
Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Japan.
Toxicol Appl Pharmacol. 2008 Sep 15;231(3):300-7. doi: 10.1016/j.taap.2008.04.020. Epub 2008 May 2.
Cytochrome P450 (CYP) 2E1 was suggested to be the major enzyme involved in trichloroethylene (TRI) metabolism and TRI-induced hepatotoxicity, although the latter molecular mechanism is not fully understood. The involvement of CYP2E1 in TRI-induced hepatotoxicity and its underlying molecular mechanism were studied by comparing hepatotoxicity in cyp2e1+/+ and cyp2e1-/- mice. The mice were exposed by inhalation to 0 (control), 1000, or 2000 ppm of TRI for 8 h a day, for 7 days, and TRI-hepatotoxicity was assessed by measuring plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and histopathology. Urinary metabolites of trichloroethanol and trichloroacetic acid (TCA) were considerably greater in cyp2e1+/+ compared to cyp2e1-/- mice, suggesting that CYP2E1 is the major P450 involved in the formation of these metabolites. Consistent with elevated plasma ALT and AST activities, cyp2e1+/+ mice in the 2000 ppm group showed histopathological inflammation. TRI significantly upregulated PPARalpha, which might function to inhibit NFkappaB p50 and p65 signalling. In addition, TRI-induced NFkappaB p52 mRNA, and significantly positive correlation between NFkappaB p52 mRNA expression and plasma ALT activity levels were observed, suggesting the involvement of p52 in liver inflammation. Taken together, the current study directly demonstrates that CYP2E1 was the major P450 involved in the first step of the TRI metabolism, and the metabolites produced may have two opposing roles: one inducing hepatotoxicity and the other protecting against the toxicity. Intermediate metabolite(s) from TRI to chloral hydrate produced by CYP2E1-mediated oxidation may be involved in the former, and TCA in the latter.
细胞色素P450(CYP)2E1被认为是参与三氯乙烯(TRI)代谢及TRI诱导的肝毒性的主要酶,尽管后者的分子机制尚未完全明确。通过比较cyp2e1+/+和cyp2e1-/-小鼠的肝毒性,研究了CYP2E1在TRI诱导的肝毒性中的作用及其潜在分子机制。将小鼠每天吸入0(对照)、1000或2000 ppm的TRI,持续8小时,共7天,并通过测量血浆丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)活性以及组织病理学来评估TRI肝毒性。与cyp2e1-/-小鼠相比,cyp2e1+/+小鼠尿中的三氯乙醇和三氯乙酸(TCA)代谢物含量显著更高,表明CYP2E1是参与这些代谢物形成的主要P450。与血浆ALT和AST活性升高一致,2000 ppm组的cyp2e1+/+小鼠出现了组织病理学炎症。TRI显著上调PPARα,其可能发挥抑制NFκB p50和p65信号传导的作用。此外,观察到TRI诱导NFκB p52 mRNA,且NFκB p52 mRNA表达与血浆ALT活性水平之间存在显著正相关,提示p52参与肝脏炎症。综上所述,本研究直接表明CYP2E1是参与TRI代谢第一步的主要P450,产生的代谢物可能具有两种相反的作用:一种诱导肝毒性,另一种保护免受毒性。由CYP2E1介导的氧化作用从TRI生成水合氯醛的中间代谢物可能参与前者,而TCA参与后者。