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目的观察养心通脉有效部位方对大鼠骨髓间充质干细胞(MSCs)体外分化为心肌样细胞的诱导作用。方法体外培养大鼠MSCs,设养心通脉有效部位方(YTF)组、5-氮胞苷(5-aza)阳性对照组和低糖DMEM组,体外诱导分化心肌样细胞;用免疫细胞化学方法观测其心肌肌丝蛋白(Desmin)、肌球蛋白重链(MHC)的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达。结果大鼠MSCs体外向心肌样细胞分化诱导后,在DMEM组中未发现Desmin和MHC阳性细胞;而YTF组和5-aza组的Desmin及MHC免疫组化染色均可见阳性免疫复合物,与DMEM组比较,YTF组和5-aza组Desmin、MHC、GATA-4 mRNA表达的相对光密度(IOD)值均明显提高(P<0.01,P<0.05),而且YTF组Desmin、MHC的IOD也明显高于5-aza组(P<0.05),但YTF组GATA-4 mRNA的IOD值与5-aza组比较无统计学差异。结论 YTF在体外可诱导MSCs向心肌样细胞分化。
Objective To observe the induction of effective fractions of Yangxin Tongmai on the differentiation of rat bone marrow mesenchymal stem cells (MSCs) into cardiomyocyte-like cells in vitro. Methods Rat MSCs were cultured in vitro and cultured in YTF group, 5-aza positive control group and low glucose DMEM group. Cardiomyocyte-like cells were induced by immunocytochemistry The myocardial myofilament protein (Desmin) and myosin heavy chain (MHC) expression were observed. The expression of myocardial transcription factor GATA-4 mRNA was detected by RT-PCR. Results Desmin and MHC positive cells were not found in the DMEM group induced by differentiation of cardiomyocyte-like cells in vitro. The positive immune complexes of Desmin and MHC were found in both YTF and 5-aza groups, The relative optical density (IOD) values of Desmin, MHC and GATA-4 mRNA in YTF group and 5-aza group were significantly increased (P <0.01, P <0.05), and the IOD of Desmin and MHC in YTF group was also significantly higher (P <0.05). However, the IOD value of GATA-4 mRNA in YTF group was not significantly different from that in 5-aza group. Conclusion YTF can induce MSCs differentiate into cardiomyocyte in vitro.