肝细胞移植治疗肝硬化的疗效

Curative effect of hepatocyte transplantation on hepafic cirrhosis

目的建立SD大鼠药物性肝硬化模型,探讨原代肝细胞移植治疗大鼠肝硬化的疗效。方法经四氯化碳(Carbon tetrachloride,CCL4)诱导大鼠肝硬化模型,将40只模型大鼠分为2组,每组20只,Ⅰ组经脾尾注射约2×107个肝细胞,Ⅱ组经脾尾注射0.4 ml Hank’s液。观察每组大鼠的存活情况及肝功能变化;HE染色观察脾脏病理变化;免疫荧光观察植入的肝细胞在大鼠脾内合成白蛋白(Serum albumin level,ALB)的功能。结果经CCL4诱导10周后,大鼠肝脏经HE染色发现假小叶形成,表明成功建立了肝硬化模型;移植术后8周,Ⅰ组大鼠存活率(75%)显著高于Ⅱ组(35%),差异有统计学意义(P<0.01);Ⅰ组大鼠的血清ALB水平较Ⅱ组明显升高(P<0.05),血氨(Serum ammonia level,AMM)水平较Ⅱ组明显降低(P<0.05);Ⅰ组大鼠移植术后1、8周,大量肝细胞在脾脏红髓中定植下来;Ⅰ组大鼠移植术后5周,植入的肝细胞在大鼠脾内具有合成白蛋白的功能。结论 CCL4能稳定诱导肝硬化动物模型;同种异体肝细胞移植能提高肝硬化大鼠的生存率,并改善肝性脑病和低蛋白血症。

Objective To establish SD rat model of drug-induced hepafic cirrhosis and investigate the curative effect of primary hepatocyte transplantation on the disease.Methods Rat model of hepafic cirrhosis was established by induction with carbon tetrachloride.Forty model rats were randomly divided into two groups.The rats in group Ⅰ were injected with 2 × 107 hepatocytes through intraplenic injection,while those in groupⅡ with 0.4 ml of Hanks solution.The rats in various groups were observed for survival rate and change of liver function,while for pathological changes of liver and spleen by HE staining,and for synthesis of albumin in transplanted hepatocytes by IFA.Results Pseudolobule was formed in liver of rats 10 weeks after induction with carbon tetrachloride as proved by HE staining.The survival rate of rats in groupⅠ was significantly higher than that in groupⅡ within 8 weeks after transplantation（P 0.01）.The serum ammonia and albumin levels of rats in groupⅠ were improved significantly（both P 0.05）.A large quantity of hepatocyes were found in splenic red pulp of rats in group Ⅰ 1 and 8 weeks after transplantation.However,scattered green fluorescence was observed in spleen of rats in group Ⅰ 5 weeks after transplantation.Conclusion Carbon tetrachloride induced hepafic cirrhosis in animal model stably.Allogenic hepatocyte transplantation increased the survival rate of rats with hepafic cirrhosis,and improved hepatic encephalopathy and hypoalbuminemia.