Research of the mechanism of non-obstructive cholestasis in severe infection and of the treatment for hepatic insufficiency

(1)有研究认为，由培养的小肝细胞构成的肝样结构中，胆小管网的不规则收缩多为不规则收缩。这种运动是通过分析体外胆小管收缩变化的特异性软应用计算出来的数字。此外，还研究了一些环境及其对膜上几种转运蛋白的影响。在这个研究计划中，已经发表了一些有趣的结果。(2)体外培养纯化自Sprague-Dawley大鼠（雄性，6-10周龄）肝脏的小肝细胞成功完成肝样组织。然后用半定量RT-PCR和免疫荧光法研究定植的构成肝样组织的小肝细胞膜上转运体的分布。将保存在冰箱中的小肝细胞反复培养，成功的结果表明，在培养开始后的4周内，定植的肝细胞略有增大，培养的肝细胞之间形成了具有胆管小梁型的肝样组织。RT-PCR分析明确观察到胆红素和胆汁酸的摄取型转运蛋白Oatp（有机阴离子转运多肽）家族、Ntcp （Na牛磺酸胆酸共转运多肽）家族、Mrp2（多重耐药相关蛋白2）家族和Besp（胆汁酸出口泵）家族的上调或其排泄型转运蛋白的上调。在培养中添加TNF-α，诱导Oatp2和CAR（组成型雄烷受体）蛋白下调或降低。(3)从我科肝切除术后切除的肝脏中分离出人小肝细胞。这些操作是在没有HBV和HCV感染的材料上进行的血清分析。在所有案例中都获得了成功的结果，但确定了结果的巨大差异。将继续进行进一步研究。少

(1)It has been suggested that the mostly irregular contractions of bile canalicular net work constituted among the hepatoid structure by cultured small hepatocytes were. This movement was presented as calculated number by the specific-soft utilization of analysis for the changes of bile canalicular contraction in vitro. Moreover, some circumstances and their influences on several transporters on membrance have been studied. Interesting results have already been published during this research schedule.(2)The completion of hepatoid tissue in vitro by cultured small hepatocyte purified from Sprague-Dawley rat (male, 6-10 week age) liver had successfully been performed. And then the distribution of transportaters localized on membrane of colonied small hepatocytes which constituted hepatoid tissue was studied by the hemi-quantitative RT-PCR and the immunofluorescence method. Preserved small hepatocytes in refregerater had been tried to culture repeatedly and the successful results showed t … More hat the colonied hepatocytes were slightly enlarged during 4 weeks after the start of culture and the hepatoid tissue with trabecular patterns of bile canallculus formed among cultured hepatocytes was organized. On the analysis of RT-PCR, the up-regulations of Oatp (organic anion transporting polypeptide) family, Ntcp (Na taurocholate cotransporting polypeptide), Mrp2 (multiple resistance associated protein 2) and Besp (bile salt export pump), which of all are the up-take transporters of bilirubin and bile acids or the excretion-type transporters of them, were definitely observed. Under the TNF-α addition in culture, the down-regulation or decreased of proteins of Oatp2 and CAR (constitutive androstane receptor), was induced.(3) Human small hepatocyte was isolated from the resected liver after hepatectomy in our department. Those procedures were performed on the material without HBV and HCV infection by serum-analysis. Successful results were experienced in all cases but wide differences in results were ascertained. Further studied would be continued. Less