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

(1)It认为在培养的小肝细胞的肝样结构中构成的胆小管网的大多数不规则收缩。通过对离体胆管收缩变化的特异性分析，将这种运动表示为计算数。此外，还研究了一些环境条件及其对几种膜上转运蛋白的影响。在这个研究计划中已经发表了一些有趣的结果。（2）从6-10周龄雄性SD大鼠肝脏中分离纯化小肝细胞，成功地完成了体外肝样组织的构建。然后用半定量RT-PCR和免疫荧光方法研究了肝样组织中小肝细胞膜上转运蛋白的分布。用冷冻箱保存的小肝细胞进行反复培养，结果表明， 关于我们 在培养开始后4周内，集落肝细胞略有增大，并在培养的肝细胞之间形成具有胆管小梁图案的肝样组织。RT-PCR分析表明，胆红素和胆汁酸的摄取转运蛋白Oatp（organic anion transporting polypeptide）家族、牛磺胆酸钠共转运蛋白Ntcp（Na taurocholate co-transporting polypeptide）、多药耐药相关蛋白2 Mrp 2（multiple resistance associated protein 2）和胆汁盐输出泵Besp（bile salt exportpump）表达上调。TNF-α诱导Oatp 2和CAR（组成型雄烷受体）蛋白表达下调或减少。(3)人小肝细胞是从我科肝切除术后切除的肝脏中分离出来的。通过血清分析对无HBV和HCV感染的材料进行这些程序。在所有情况下，都取得了成功的结果，但确定了结果的广泛差异。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