脐血干细胞与骨髓干细胞联合移植治疗大鼠急性肝衰竭实验研究

Experimental study of transplantations of umbilical cord blood stem cells combined with bone marrow stem cells for rats with acute liver failure

目的观察脐血干细胞和自体骨髓干细胞共同移植治疗D-半乳糖苷所致急性肝衰竭大鼠的疗效。方法采集大鼠脐血和骨髓中单个核细胞,以促肝细胞生长因子和干细胞因子培养3w,采用免疫细胞化学法检测白蛋白和甲胎蛋白表达情况;以D-半乳糖苷腹腔注射法建立大鼠急性肝衰竭模型,24 h后每日经鼠尾静脉分别注入脐血干细胞,或骨髓干细胞,或混合的脐血干细胞和骨髓干细胞,对照组注射等量生理盐水,治疗7d,观察4组大鼠存活率、肝功能和肝组织病理学变化。结果在促肝细胞生长因子及干细胞因子的诱导下,脐血干细胞和骨髓干细胞可以在体外扩增并分化为肝细胞;脐血干细胞移植、骨髓干细胞移植和联合细胞移植组大鼠9d存活率分别为55.6%、50.0%和77.8%,均明显高于生理盐水对照组(16.7%,P<0.01);联合移植组存活率高于任何一种单纯干细胞移植(P<0.01)。结论脐血干细胞和骨髓干细胞移植对大鼠急性肝损伤有一定的保护作用,两者联合移植有协同作用。

Objective To observe the therapeutic effects of co-transplanted umbilical cord blood stem cells and bone marrow stem cells for rats with acute liver failure induced by D-galactose. Methods Mononuclear cells isolated from umbilical cord blood and bone marrow of rats were cultured in medium containing hepatocyte growth factor（HGF） and stem cell factor（SCF） for 3 weeks,and the expression of hepatocyte markers,such as AFP and ALB,were detected by immunocytochemistry;A rat model of acute liver failure was established by D-galactose in-jection （1.4 g.kg-1,intraperitoneally） for 24 h. Rats with acute liver failure were subjected to transplantation of bone marrow stem cells,umbilical cord blood stem cells,and umbilical cord blood stem cell mixed with bone mar-row stem cell,or an equal amount of saline per day for seven days. The survival rates,liver function and patho-logical changes in liver were studied. Results Bone marrow stem cells and umbilical cord blood stem cells were successfully proliferated and differentiated into hepatocytes in vitro by stimulation of HGF and SCF. The 9-day survival rates of rats with acute liver failure in umbilical cord blood stem cell,bone marrow stem cell and mixed stem cell transplantation group were 55.6%,50.0% and 77.8%,respectively,all of which were markedly higher than that in rats treated with saline（16.7%,P〈0.01）;The 9-day survival rate of rats in mixed cell transplantation group was significantly higher than that of rats in umbilical cord blood stem cell or bone marrow stem cell alone （P〈0.01）. Conclusions Bone marrow stem cell and umbilical cord blood stem cell transplantation improve liver function of rats with acute liver failure,and combinational transplantation of these two cells is efficient.