Ilyin G P, Rissel M, Malledant Y, Tanguy M, Guillouzo A
INSERM U 49, Unité de Recherches Hépatologiques, Hôpital de Pontchaillou, Rennes, France.
Biochem Pharmacol. 1994 Aug 3;48(3):561-7. doi: 10.1016/0006-2952(94)90286-0.
Biotransformation of anaesthetic halothane by cytochrome P450-dependent monooxygenases resulted in the production of reactive intermediate trifluoroacetyl (TFA) halide, capable of covalently binding to hepatocyte proteins. TFA-modified liver proteins can act as antigens and are implicated in the pathogenesis of halothane hepatitis in humans. The aim of this study was to investigate the formation of TFA-neoantigens in halothane-treated primary cultures of adult human hepatocytes and to evaluate the usefulness of this in vitro model for studying immune-mediated halothane hepatotoxicity. Cultured human hepatocytes were incubated with halothane under constant temperature, atmosphere and anaesthetic concentration conditions. The results obtained show that halothane-treated hepatocytes isolated from seven different donors produced TFA-antigens as detected by immunocytochemical and western immunoblot analysis using rabbit anti-TFA antiserum. TFA-adducts were localized mainly in the endoplasmic reticulum and in small amounts on the plasma membrane of parenchymal cells. By immunoblotting, several neoantigens, with molecular masses from 42 to 100 kDa, were detected in halothane-exposed hepatocytes. These observations are consistent with the formation of TFA-adducts through metabolism of the anaesthetic and suggest that primary cultures of human hepatocytes represent a suitable in vitro model to study the pathogenesis of immune-mediated halothane hepatotoxicity.
细胞色素P450依赖性单加氧酶对麻醉药氟烷的生物转化导致反应性中间体三氟乙酰（TFA）卤化物的产生，该卤化物能够与肝细胞蛋白共价结合。TFA修饰的肝蛋白可作为抗原，并与人类氟烷性肝炎的发病机制有关。本研究的目的是研究在氟烷处理的成人原代肝细胞培养物中TFA新抗原的形成，并评估该体外模型在研究免疫介导的氟烷肝毒性方面的实用性。将培养的人肝细胞在恒定温度、气氛和麻醉浓度条件下与氟烷孵育。所得结果表明，使用兔抗TFA抗血清通过免疫细胞化学和western免疫印迹分析检测到，从七个不同供体分离的经氟烷处理的肝细胞产生了TFA抗原。TFA加合物主要定位于内质网，在实质细胞的质膜上有少量分布。通过免疫印迹法，在暴露于氟烷的肝细胞中检测到几种分子量在42至100 kDa之间的新抗原。这些观察结果与通过麻醉药代谢形成TFA加合物一致，表明人肝细胞原代培养物是研究免疫介导的氟烷肝毒性发病机制的合适体外模型。
