脐血干细胞移植治疗肝功能衰竭及心肌损害的临床与实验研究

Clinical and experimental study of the therapeutic effect of umbilical cord blood stem cell transplantation on liver failure and heart damage in severe viral hepatitis patients

目的探讨脐血干细胞移植(UCBSCT)治疗合并心肌损害的重型肝炎肝功能衰竭的疗效并与无心肌损害的重型肝炎患者及成人新鲜血浆的疗效进行比较。观察UCBSCT治疗大鼠肝功能衰竭的效果。方法83例有(或无)心肌损害的重型肝炎患者分为4组,各组分别采用UCBSCT或成人新鲜血浆治疗。观察各组患者治疗前后肝功能及心肌酶学指标的变化并进行比较。动物实验用四氯化碳诱导急性肝损伤模型,分别自大鼠尾静脉输注等渗盐水、脐血清及脐血干细胞。分批将鼠杀死并取肝脏组织用免疫组织化学方法检测人肝细胞甲胎蛋白和白蛋白的表达及聚合酶链反应(PCR)检测人DNA的存在。结果UCBSCT改善肝脏功能的疗效明显优于成人新鲜血浆。心肌损害对于UCBSCT治疗重型肝炎的疗效无明显影响。UCBSCT对于患者的心肌损害也有一定的改善作用。动物实验中输注脐血干细胞后21d和1个月的大鼠肝脏中可见人甲胎蛋白和白蛋白表达;PCR方法可以扩增出人特异的DNA片段。结论UCBSCT对于重型肝炎具有较好的疗效,同时可以减轻患者合并的心肌损害。动物实验提示脐血干细胞可以减轻肝脏损害及促进肝细胞再生,人脐血干细胞可以在急性肝损伤的SD大鼠体内分化为肝细胞。

Objectives To investigate and compare the therapeutic effect of umbilical cord blood stem cell transplantation (UCBSCT) or adult fresh plasma in severe viral hepatitis liver failure with/without heart damage, and to study the effect of UCBSCT on liver lesions in rats. Methods 83 severe hepatitis patients with/without heart damage were included in the study between January 1994 and June 2003. The patients were treated with UCBSCT or given adult plasma transfusions. The therapeutic effect was evaluated by serial determination of liver function and myocardium enzymes in all patients before and after the treatment. The model of experimental hepatic failure was constructed in SD rats by injecting carbon tetrachloride. Then, the rats were given normal saline, neonate cord blood serum or neonate cord blood stem cells respectively. The expression of human AFP and Alb in SD rat livers was detected by immunohistochemistry; and human special DNA was detected by PCR. Results The UCBSCT group had much better effects in the improvement of liver function than the adult plasma group had, no matter whether the patients had heart damage or not. Moreover, UCBSCT can decrease heart impairment of the patients. The animal experiment demonstrated that AFP and Alb positive cells were present in the neonate cord blood stem cell group after 21 days and 1 month; human special DNA was detected by PCR in these SD rat livers. Conclusion UCBSCT displayed good therapeutic effects on severe viral hepatitis and improvement of heart injury of the patients. The rat liver immunohistochemistry indicated that neonate cord blood stem cell application can decrease the liver damage and increase hepatocellular regeneration. Human umbilical cord blood stem cells can differentiate into liver cells in acute damaged SD rat livers.