肝细胞系Chang liver细胞移植对改善急性肝衰竭存活率的作用的研究

目的 通过体外实验及体内急性肝衰竭模型验证其肝细胞功能,为生物人工肝支持系统寻找一种可用的细胞源。方法 通过Western-Blotting和激光共聚焦检测Chang Liver细胞的肝细胞特异性蛋白表达,在体外评估其肝细胞功能。采用RT-PCR从基因水平检测肝细胞功能相关基因的表达情况。采用大鼠90%肝切除术诱导急性肝衰竭模型(ALF)。取生长良好的Chang Liver细胞移植入脾脏内,并于移植后24 h行90%肝切除术诱导急性肝衰竭,每隔24 h采血1次,测定肝功能指标变化情况。结果 在体外,Chang Liver细胞表达肝细胞特异性标志蛋白,如白蛋白(ALB)、葡糖醛酸基转移酶(UGT)及细胞色素酶P450(CYP3A4)。RT-PCR 同样可检测到相关基因在mRNA水平的表达。在体内,实验组接受Chang Liver细胞移植,其长期生存率达到40%,而对照组的生存率为0。移植Chang Liver细胞的大鼠的肝功能在急性肝衰竭后24 h较对照组获得明显的改善,表现为血样标本中谷丙转氨酶(ALT),谷草转氨酶(AST),胆红素和碱性磷酸酶(ALP)等肝功能指标的下降。结论 Chang Liver细胞可像正常肝细胞一样表达功能蛋白,明显改善急性肝衰竭大鼠的肝功能,改善存活率,可以作为生物人工肝支持系统较好的细胞源。

Transplantation of a hepatocye cell line Chang liver cell improves the survival of acute liver faliure

Objective To verify the function of hepatocytes by in vitro experiments and in vivo acute hepatic failure model and search for a novel cell source for liver support system.Methods the expressions of specific hepatocyte markers were detected by western-blotting and laser confocal microscopy to evaluate the hepatocytic function of Chang Liver cells in vitro. Related gene expression of hepatic function was detected by RT-PCR.The acute liver failure (ALF) model was established by 90% partial hepatectomy.Chang Liver cells were intra-splenically implanted 24 hours before the surgery and the serum samples were collected per 24 hours to test the liver functions. And the survivals were assessed.Results In vitro, conspicuous expression of hepatocytic markers,such as albumin (ALB),uridine diphosphate glucuronosyltransferase (UGT) and cytochrome P450 3A4 (CYP3A4) in Chang Liver cells.RT-PCR also exhibited the expression of related genes at mRNA level.In vivo,the survival of rats receiving Chang Liver cells was significantly improved and reached 40% against 0% in controls.Liver functions of rats receiving Chang Liver cells were also significantly improved 24 h after ALF,which was evidenced by the decreased levels of alanine transaminase (ALT), aspartate aminotransferase (AST), bilirubin and alkaline phosphates (ALP) in blood samples of rats implanted with Chang liver cells.Conclusion Chang Liver cells express hepatocytic markers and significantly improve liver function and survival after ALF, which can serve for liver support systems cells.