Satoh H, Fukuda Y, Anderson D K, Ferrans V J, Gillette J R, Pohl L R
J Pharmacol Exp Ther. 1985 Jun;233(3):857-62.
The fulminant hepatotoxicity caused by halothane has been thought to have an immunological basis because this toxicity occurs most often after repeated administration of halothane and because sera from patients recovering from severe halothane hepatotoxicity contain antibodies that bind to the surface membranes of hepatocytes of rabbits treated with halothane. In order to determine whether the major reactive metabolite of halothane, trifluoroacetyl halide, covalently binds to hepatocytes, we have developed specific and sensitive peroxidase enzyme-linked immunosorbent assays and an indirect immunofluorescence staining method for identifying trifluoroacetylated (TFA)-hepatocytes. Liver sections prepared from rats at 4 hr after halothane administration were stained preferentially in the centrilobular region with anti-TFA serum whereas livers of control rats showed no staining. The specificity of the assay for the TFA group was confirmed by the complete inhibition of the staining with 200 microM N-epsilon-TFA-L-lysine in the diluted antiserum. On the other hand, 2 mM halothane or L-lysine did not inhibit this staining. Moreover, treatment of rats with deuterated halothane resulted in significantly less staining than did halothane. At 24 hr after halothane administration, hepatocytes isolated and stained by indirect immunofluorescence showed a linear and granular pattern on their surface membranes. These results indicate that trifluoroacetyl halide either reacts directly with constituents of the plasma membranes or with other cellular components which become incorporated into the plasma membranes.
氟烷引起的暴发性肝毒性被认为具有免疫学基础，因为这种毒性最常发生在反复使用氟烷之后，而且从严重氟烷肝毒性中恢复的患者血清中含有能与用氟烷处理的兔肝细胞表面膜结合的抗体。为了确定氟烷的主要反应性代谢产物三氟乙酰卤是否与肝细胞共价结合，我们开发了特异性和灵敏的过氧化物酶联免疫吸附测定法以及间接免疫荧光染色法来鉴定三氟乙酰化（TFA）肝细胞。氟烷给药后4小时从大鼠制备的肝切片在小叶中心区域被抗TFA血清优先染色，而对照大鼠的肝脏则无染色。在稀释的抗血清中加入200微摩尔N-ε-TFA-L-赖氨酸可完全抑制染色，从而证实了该测定法对TFA基团的特异性。另一方面，2毫摩尔氟烷或L-赖氨酸不能抑制这种染色。此外，用氘代氟烷处理的大鼠染色明显少于氟烷处理的大鼠。氟烷给药后24小时，通过间接免疫荧光分离和染色的肝细胞在其表面膜上呈现线性和颗粒状模式。这些结果表明，三氟乙酰卤要么直接与质膜成分反应，要么与其他并入质膜的细胞成分反应。
