Objective To investigate the effects of different therapeutic formulas on the differentiation of bone marrow mesenchymal stem cells ( BMSC) to cardiomyocyte-like cells in rat myocardium of in vitro micro-environment. Methods 40 SD rats randomly divided into blood-activating stasis-resolving group, qi supplementing blood-activating group, qi supplementing group, and blank group were administered o-rally Chinese medicinals to obtain madicated serum;another 60 SD rats were used to extract, incubate, i-solate and purify BMSC;SD newborn rats were used to extract and incubate myocardial cell, then to es-tablish an external myocardial micro-environment. MTT assay was used to select the optimal compounding n concentration of each treatment group. Flow cytometry was used to measure the proliferation of CD105 +-BMSC of each treatment group. Immunocytochemical method was used to test the CD105 +-BMSC’ s ex-pression of myocardium protein CTNT, GATA-4 while immunofluorescence technique was used to meas-ure the expression of MHC. Results The doubling time of CD105 +-BMSC of each treatment group was 24 h. The cells numbers in blood-activating and stasis-resolving group were significantly higher than those in control group and other treatment groups from the 2nd day, and remained steady. All the proliferation index of blood-activating and stasis-resolving group at every time point were the highest except the 1 st day. The number of cell division was also significantly more than other groups: qi-supplementing and blood-activating group, qi-supplementing group, blood activating and stasis resolving group. From the 2nd day to 5th day, the expressions in the blood-activating stasis-resolving group, qi-supplementing blood-activating group, qi-supplementing group increased markedly with statistical significance compared with the blank group ( P<0 . 05 ) , blood-activating stasis-resolving group was superior to the other two groups(P<0. 05). There were no statistical significance between the qi-supplementing blood-activating group and qi-supplementing group ( P>0 . 05 ) . Conclusion Under the external mimic myocardial mi-cro-environment condition, all therapeutic formulas could promote CD105 +-BMSC ’ s differentiation. They also could promote CD105 +-BMSC ’ s metaplasia to the cardiomyocyte-like cells, however, the blood-activating stasis-resolving medicinal appears to be most effective.%目的：探讨体外心肌微环境下不同治法方药诱导骨髓间充质干细胞( BMSC )向心肌样细胞分化的差异性。方法40只成年SD大鼠随机分为活血化瘀组、益气活血组、益气组、空白组,给相应药物后制备含药血清；另再从60只正常SD大鼠提取、培养及分离纯化BMSC；同时提取、培养SD新生鼠心肌细胞,建立体外模拟大鼠心肌微环境模型。用MTT法选择各给药组的最佳倍比浓度；流式细胞仪检测BMSC中CD105阳性细胞( CD105+-BMSC)分裂增殖情况；免疫细胞化学法检测心肌肌钙蛋白T(CTNT)、心肌细胞转录因子4(GATA-4)分化情况；免疫荧光技术检测心肌特异性肌球蛋白重链(MHC)表达。结果各用药组CD105+-BMSC的群体倍增时间为24 h,活血化瘀组的细胞数于第2天开始明显高于其他用药组和对照组且呈持续状态。活血化瘀组除第1天外各时间点的增殖指数均为最高值,分裂代次明显多于其他组。第2、3、4、5天,活血化瘀组、益气活血组、益气组的MHC、CTNT和GATA-4表达量比空白组明显增加( P<0.05)；活血化瘀组明显优于益气活血组、益气组(P<0.05)；益气组与益气活血组差别无统计学意义(P>0.05)。结论在体外模拟心肌微环境的条件下,活血化瘀法与益气活血法、益气法均有促进CD105+-BMSC分裂并向心肌样细胞分化的作用,其中活血化瘀法效果最为明显。
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