人胚胎肝细胞移植治疗大鼠急性肝衰竭的效果分析

Effective analysis on treatment of rat acute liver failure by human fetal hepatocyte transplantation

目的探讨人胚胎肝细胞移植治疗急性肝衰竭(ALF)的疗效及能否成为移植细胞源。方法采用四甲基偶氮唑盐法检测人胚胎肝细胞增殖情况;免疫细胞化学检测其ALB、细胞角蛋白-18(CK-18)的表达;D-氨基半乳糖(D-gal)药物诱导ALF的实验动物模型;人胚胎肝细胞移植治疗ALF的动物模型,包括移植组与对照组在肝功能指标[ALT、AST、碱性磷酸酶(ALP)、总胆红素(TBIL)]的差异性比较。免疫组织化学法检测植入脾脏中的人胚胎肝细胞ALB及CK-18的表达。结果培养第5天左右细胞数量达到最高峰,每天完成4～5次分裂,胚胎肝细胞的生长曲线呈抛物线型。免疫细胞化学检测提示其具有表达ALB、CK-18的功能;D-gal 1.6 g/kg腹腔注射72 h后大鼠ALF模型制备成功;移植第3～5天后,移植组与对照组比较,肝功能(ALT、AST、ALP、TBIL)指标差异有统计学意义(P<0.01)。植入脾内的肝细胞具有分泌并表达CK-18、ALB的功能。结论人胚胎肝细胞脾内移植能有效治疗ALF,并可能成为肝细胞移植的靶细胞源。

Objective To investigate the effect of treating rat acute liver failure and alternative sources of cells for transplantation by human fetal hepatocyte transplantation Methods The proliferation,expression of albumin and cytokeratin-18 of human fetal hepatocyte were detected by MTT and immunocytochemistry.The animal models of rat acute liver failure were made by D-galactosamine.Hepatic functions,such as alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin were measured between the treated and the control groups.The expression of albumin and cytokeratin-18 of human fetal hepatocyte transplanted in spleen were detected by immunohistochemistry.Results The cell population reached a peak,when human fetal hepatocyte were cultivated for 5 d.The cell division had 4-5 times per day,its growth curve present parabola.Immunocytochemistry revealed the positive expression of albumin and cytokeratin-18 of human fetal hepatocyte.The models of rat acute liver failure were made succeedly in 72 h by D-galactosamine 1.6 g/kg intraperitoneal injection.Alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin were measured and there were significant difference between the treated and the control groups（P0.01）.Immunohistochemistry revealed the positive expression of albumin and cytokeratin-18 of human fetal hepatocyte transplanted in spleen.Conclusion These findings demonstrate the treating of rat acute liver failure by human fetal hepatocyte transplantation is effective and alternative sources of cells for transplantation.