骨髓间充质干细胞移植对小鼠急性肝功能衰竭的治疗作用

目的:观察骨髓间充质干细胞(MSCs)移植对小鼠急性肝功能衰竭(ALF)的治疗作用，为MSCs的临床应用提供理论依据。方法:分离、纯化、扩增、鉴定小鼠MSCs，用CCl4灌胃制备稳定的小鼠ALF模型，将小鼠随机分为实验组和对照组，MSCs经尾静脉移植入实验组小鼠体内，对照组小鼠注入等量生理盐水，观察2组小鼠的活动、对外界反应及存活情况，检测2组小鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)及总胆红素(TBIL)，并进行病理学检测，免疫组织化学染色法检测肝脏白蛋白（ALB）表达。结果：流式细胞术检测，分离培养的MSCs 75.27%处于G0-G1期，而且MSCs强表达CD44，表达率为88.71%，基
本不表达造血细胞表面标志物CD34。将MSCs移植入ALF小鼠体内后，实验组小鼠术后7 d生存率为65%，明显高于对照组（25%）（P<0.01）。实验组与对照组小鼠血清ALT、AST及TBIL水平均明显升高，但实验组明显低于对照组，各时间点2组间比较差异有统计学意义（P<0.01）。实验组小鼠肝细胞坏死程度及炎细胞浸润等明显轻于对照组；7 d后实验组小鼠病理组织学改善情况较对照组明显。免疫组织化学检测，7 d后实验组小鼠肝组织中ALB表明显高于对照组。结论:骨髓MSCs移植能明显改善ALF小鼠的肝功能，提高其生存率，提示骨髓MSCs移植对小鼠ALF具有明显的治疗作用。

Therapeutic effect of transplantation of mesenchymal stem cells on acute liver failure in mice

Objective To observe the therapeutic effect of transplantation of mesenchymal stem cells(MSCs) on acute liver failure(ALF) in mice,and to provide a theoretical basis for further clinical application of MSCs.Methods The MSCs of mice were isolated,cultivated,purified and identified in vitro.The model of ALF induced by CCl4 was established.The ALF mice were randomly divided into control and treatment groups.The MSCs were transplanted into tail veins of mice in treament group,and saline was transfused into the tail veins of mice in control group.The levels of serum ALT,AST and TBIL were determined.The pathological changes of liver tissues were observed by microscope after stained with HE.The survival rate was defined as the percentage of surviving animals at any specific time points.Results The cultured MSCs at G0-G1 stage accounted for 75.27% detected by flow cytometry.The expression of CD44 in MSCs was positive(88.71%),while the expression of CD34 was negative.After the MSCs were transplanted to ALF model rats,the survival rate in treatment group(65%) was higher than that in control group(25%).The levels of serum ALT,AST and TBIL were decreased after transplantation of MSCs compared with control group(P<0.01).The degrees of liver cell necrosis and inflammatory cell infiltration in treatment group were lighter than those in control group on day 1 and 2.And the pathological improvement of liver in treatment group was more evident than that in control group on day 7.The expression level of ALB in treatment group was higher than that in control group on day 7 detected by immunohistochemistry.Conclusion Transplantation of MSCs can improve liver function and increase survival rates obviously of mice with ALF,which indicates that transplantation of MSCs has an obviously therapeutic effect on ALF in mice