黄芪注射液与5-氮杂胞苷体外诱导大鼠骨髓间充质干细胞分化为心肌细胞的研究

目的研究黄芪注射液与5-氮杂胞苷（5-Aza）对大鼠骨髓间充质干细胞（mesenchymal stem cells，MSCs）分化为心肌细胞的影响。方法在无菌条件下从成年大鼠胫骨骨髓分离出MSCs。以1：3的比例传代，取第3代的细胞．接种后随机将MSCs分为黄芪注射液诱导分化组、5-Aza诱导分化组、黄芪注射液预处理后5-Aza诱导分化组。分别以黄芪注射液、5-Aza及黄芪注射液＋5-Aza进行诱导，用相差显微镜观察各纽细胞形态变化；免疫细胞化学鉴定MSCs表面标志物及心肌特异性肌钙蛋白I。结果各组诱导后细胞形态发生改变，细胞之问形成连接，排列方向趋于一致，免疫组化检测结果显示肌钙蛋白I（eTnI）表达均阳性。黄芪注射液组与5-Aza组相比其诱导率无明显区别．但黄芪注射液＋5一Aza组阳性细胞比率均高于5-Aza组及黄芪注射液组．差异具有统计学意义（P〈0．05）。结论黄芪注射液可体外诱导MSCs分化为心肌样细胞，采用黄芪注射液预处理后5-Aza诱导分化的方法优于单纯采用黄芪注射液或5-Aza谤导法。

Study of astragalus mongholicus injection and 5-Aza on bone marrow mesenchymal stem cells to induce cardiomyocytes in rats in vitro

Objective To study the effects of on mesenchymal stem cells（MSCs） to induce cardiac adult rats and purified by expanded culturing. astragalus mongholicus injection and 5-Ata myocyte. Methods MSCs were isolated from The third passage MSCs were treated by demethylating agent astragalus mongholicus injection, 5-Aza and jection with 5-Aza. The cardiomyogenic ceils were identified with astragalus mongholicus inphase contrast microscope. The cardiac Troponin I （cTnI） and facial identification were identified by immunohistochemistry. Results The positive expression of CD44 was ceil surface antigen identified in all groups; after the cultured treatment, the shapes of cells was changed; the connections between the cells were formed; the arraying direction of the ceils was similar; texpression of cTnI was positive. Com- pared with the injection group and 5-Ata group, CD44 positive in the injection with 5-Ata group were higher （P〈0.05）. Conclusion MSCs differentiating into cardiacmyocyte. Astragalus mongholicus injection might induce