北京中医药大学学报Issue(2)：106-112,7.DOI:10.3969/j.issn.1006-2157.2016.02.004
体外心肌微环境下益气活血化瘀方药诱导骨髓间充质干细胞向心肌样细胞分化的研究∗
Effects of qi supplementing, blood activating, and blood stasis resolving medicinals on differentiation of bone mesenchymal stem cells to cardio-myocyte-like cells in microenvironment of rat myocardium：a laboratory study in vitro
摘要
Abstract
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 <br> 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.关键词
微环境/骨髓间充质干细胞/增殖/分化/大鼠Key words
Ker words：micro-environment/bone marrow mesenchymal stem cells/proliferation/differentiation/rats分类
医药卫生引用本文复制引用
张金生,张宝霞,杜梅梅,朱慧芳,张阳阳..体外心肌微环境下益气活血化瘀方药诱导骨髓间充质干细胞向心肌样细胞分化的研究∗[J].北京中医药大学学报,2016,(2):106-112,7.基金项目
国家自然科学基金项目(No． H2708/81072798) （No． H2708/81072798）
