黄芪甲苷联合丹参酮Ⅱa诱导骨髓间充质干细胞向心肌样细胞分化

Combination of tanshinone IIa and astragaloside induces bone marrow mesenchymal stem cells differentiating into myocardium-like cells

背景:黄芪甲苷和丹参酮Ⅱa是传统中医药治疗心肌缺血的有效成分,是否可参与骨髓间充质干细胞向心肌样细胞分化的过程呢?目的:观察黄芪甲苷联合丹参酮Ⅱa对体外诱导骨髓间充质干细胞向心肌样细胞分化作用的影响。方法:应用MTT法分别测定黄芪甲苷和丹参酮Ⅱa的最大无毒浓度,确定两者联合诱导骨髓间充质干细胞分化为心肌样细胞的剂量。将分离纯化的骨髓间充质干细胞分5组进行诱导:黄芪甲苷组、丹参酮Ⅱa组、丹参酮Ⅱa+黄芪甲苷组、5-氮胞苷组、空白对照组,ELISA方法观察体外诱导后骨髓间充质干细胞缝隙连接蛋白43和肌钙蛋白表达变化。结果与结论:诱导后丹参酮Ⅱa+黄芪甲苷组、丹参酮Ⅱa、黄芪甲苷、5-氮胞苷组肌钙蛋白、缝隙连接蛋白43表达水平均高于正常对照组(P<0.01),丹参酮Ⅱa+黄芪甲苷组高于丹参酮Ⅱa、黄芪甲苷组(P<0.01)。丹参酮Ⅱa+黄芪甲苷组与5-氮胞苷组缝隙连接蛋白43表达水平差异无显著性意义(P>0.05)。结果提示黄芪甲苷联合丹参酮Ⅱa可以诱导骨髓间充质干细胞向心肌样细胞方向转化,而其联合作用高于单一成分的作用。

BACKGROUND： Astragaloside and tanshinone Ila are the effective components of traditional Chinese medicine treatment of myocardial ischemia, and its role in bone marrow mesenchymal stem cells differentiation into myocardium-like cells remains unclear. OBJECTIVE： To investigate the effect of tanshinone lia and astragaioside on the differentiation of bone marrow mesenchymal stem cells into myocardium-like ceils. METHODS： The maximal non-toxic concentrations of tanshinone Ila and astragaloside were measured using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay, to define the dose of the two in the induced differentiation of bone marrow mesenchymal stem cells into myocardium-like cells. The isolated and purified bone marrow mesenchymal stem cells weredivided into five groups： astragaloside group, tanshinone Ila group, astragaloside ＋ tanshinone Ila group, .5-azacitidine group, and blank control group. The expression of gapjunction connexin 43 and troponin was determined with enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION： The expression of gap junction connexin 43 and treponin in astragaloside group, tanshinone Ila group, astragaloside ＋ tanshinone Ila group, 5-azacitidine group was higher than that in blank control group （P 〈 0.01 ）. The astragaloside ＋ tanshinone Ila group showed a higher expression of gap junction connexin 43 and troponin than astragaloside group and tanshinone Ila group （P 〈 0.01）. There was no significant difference in the expression of gap junction connexin 43 and troponin between astragaloside ＋ tanshinone lia group and 5-azacitidine group （P 〉 0.05）. A combined use of astragaloside and tanshinone Ila can induce bone marrow mesenchymal stem cells to differentiate into myocardium-like cells, and their joint role is better than the role of a single ingredient.