The study of cholestasis using immunoelectron microscopy and rapid freezing deep-etching replica

用免疫荧光、透射电子显微镜和免疫电子显微镜观察了不同胆汁淤积模型大鼠肝细胞骨架的变化。以鬼臼乙素、细胞激活素B和胆汁淤积因子处理的大鼠为肝内胆汁淤积症模型和梗阻性黄疸模型，用KatSuma等人的方法处理的大鼠肝组织中可见肝周鞘(PS)。结扎胆管后，胆管结扎组微丝(MFS)和中间丝(IF)增多，PS扩张增厚。结扎胆管的大鼠出现局灶性PS断裂，胆管再通后PS断裂消失。我们还用8号和18号抗细胞角蛋白(CK)的单抗(Moll‘s分级)研究了胆汁淤积过程中肝细胞内IF的异质性，并观察了肝细胞增生性结节中IF的变化。免疫荧光和免疫电子显微镜显示，细胞角蛋白多肽在肝癌发生过程中的染色模式发生了变化，这可能有助于肝细胞癌的早期发现。

We investigated the cytoskeletal changes of hepatocytes in various kinds of cholestatic rat models using immunofluorescence, transmission electron microscope(TEM) and immunoelectron microscope(IEM). The rats treated with phalloidin, cytokalasin B and cholestatic factor were used as the model of in- trahepatic cholestasis and the rats as the model of obstructive jaundice.Pericanalicular sheath(PS) was clearly demonstrated by TEM in the liver tissues treated by the method of Katsuma et al. Almost all filaments made up PS were identified as intermediate filaments(IFs) by IEM.In the phalloidin treated and bile duct ligated rats, microfilaments(MFS) and IFs increased in amounts and PS was dilated and thickened. The focal disruption of the PS was observed in the bile duct ligated rats, which disappeared after recanalization of the bile ducts. These results suggest that IFs increase in amounts according to the increase of MFs in cholestasis.We also investigated the immunological hetetogeneity of IFs in hepatocytes using a few kinds of monoclonal anti cytokeratin(CK) antibodies including No.8 and No. 18(Moll's classification) and the change of IFs of hepatocytes was noted in the hyperplastic nodules in the experimental model of hepatocacinogenesis. Immunofluorescence and immunoelectron microscopy demonstrated that the staining pattern of cytokeratin polypeptides changed in the process of hepatocarcinogenesis, which might be useful in the early detection of hepatocellular carcinoma.