大鼠间充质干细胞在小鼠肝损伤中的作用

The Role of Rat Mesenchymal Stem Cells in Repairing Mice Liver Damage

目的研究大鼠骨髓间充质干细胞(MSC)在四氯化碳(CCl4)导致小鼠肝损伤中的作用,为临床进行细胞移植治疗肝脏疾病提供新策略。方法无菌条件下取雄性Wistar大鼠骨髓,分离培养及流式细胞仪检测MSC;采用CCl4制作雌性BALB/c小鼠肝损伤模型,肝损伤小鼠随机分为MSC移植组和肝损伤对照组。MSC移植组在CCl4损伤后立即经尾静脉注入大鼠MSC,6周后处死小鼠,检测小鼠肝损伤程度和大鼠MSC植入情况。结果流式细胞仪结果显示:第3代大鼠MSC CD45-/CD90+细胞为94.54%,CD45-/CD44+细胞为91.25%;肝组织学观察显示:MSC移植组肝损伤修复程度明显好于肝损伤对照组;PCR方法证实只有MSC移植组肝内有大鼠Y染色体阳性细胞存在。结论体外分离培养及扩增的大鼠MSC移植到CCl4肝损伤小鼠体内,可参与肝损伤修复,减轻CCl4导致的肝损伤。

Objective To investigate the repairing rat bone marrow in liver damage caused by carbon role of mesenchymal stem cells （MSC） from tetrachloride （CCl4）, and to provide a new method for cell transplant to treat liver disease. Methods Bone marrow was collected from male Wistar rat under sterile condition. MSCs were isolated, cultured and identified by using flow cytometry. Female BALB/c mice were challenged with CCl4 to make liver damage models. The mice with liver damage were randomly divided into MSC transplanted group and control group. In the MSC transplanted group, MSCs were infused immediately through the tail veins after CCl4 treated. After six weeks, liver damage and rat cells engraftment were assessed. Results According to flow cytometry assay, at the 3rd passage MSC, the ratio of CD45^- / CD90^＋ cells was about 94. 54%, the ratio of CD45^- / CD44^＋ cells was about 91. 25%; Histological examination indecated that liver damage recovery of the MSC transplanted group was much better than that of the control group; PCR analysis revealed that rat Y-chromosome-positive cells was founded in only MSC transplanted group. Conclusion Rat MSC can be isolated, cultured and expanded in vitro. Transplanted to CCl4-treated mice. They can play a pole in liver damage recovery and ameliorate liver damage.