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

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

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组细胞形态变化;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,细胞之间形成连接,排列方向趋于一致,免疫组化检测结果显示肌钙蛋白I(cTnI)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-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诱导法。

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