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

(1)提示培养的小肝细胞在类肝样结构中所构成的胆小管网状收缩主要是不规则的。通过对体外胆小管收缩变化的特殊软件分析，将这一运动表示为计算数。此外，还研究了一些情况及其对膜上几种转运蛋白的影响。(2)从SD大鼠(雄性，6-10周龄)肝脏中提纯培养的小肝细胞，成功地完成了类肝组织的体外构建。然后用半定量RT-PCR和免疫荧光方法研究定植的小肝细胞膜上转运体的分布。将保存在培养瓶中的小肝细胞进行反复培养，成功的结果是…。培养开始后4周内，集落的肝细胞稍有增大，并在培养的肝细胞间形成具有胆小梁结构的类肝样组织。通过RT-PCR分析，明确了OATP(有机阴离子转运多肽)家族、NTCP(牛磺胆酸钠共转运多肽)、MRP2(多药耐药相关蛋白2)和BESP(胆盐输出泵)等胆红素和胆汁酸摄取转运体或排泄型转运体的表达上调。在加入肿瘤坏死因子-α的培养条件下，OATP2和CAR的蛋白表达下调或减少。(3)从我科肝切除后切除的肝组织中分离出人小肝细胞。这些手术都是通过血清分析对没有感染乙肝病毒和丙型肝炎病毒的材料进行的。所有病例都获得了成功的结果，但结果之间存在很大差异。将继续进行进一步的研究。较少

(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