动物实验,骨髓间充质干细胞 z-bo 2020-08-27 480

　　Objective: To investigate the effect of Jiawei Danshen Decoction on the differentiation of bone marrow mesenchymal stem cells BMSCs into cardiomyocyte-like cells in vitro.

　　Method: Obtain rat bone marrow mesenchymal stem cells by adherence method, and detect cell surface antigens by flow cytometry. The MTT method was used to detect the maximum non-toxic concentration of Jiawei Danshen Decoction on bone marrow mesenchymal stem cells induced by the third generation of cells: normal control group, Jiawei Danshen Decoction group, 5-azacytidine group, and Jiawei Danshen Decoction. + 5-azacytidine group, using Western blotting to detect cell binding protein (desmin), α-myosin heavy chain (α-MHC), and detect the expression of cardiac troponin T (cTnT) protein in each group, RT detects GATA-4 -PCR, Gap connexin 43 (CX43) and Nkx2-5 mRNA expression.

　　"Result: The positive rate of CD90 on the surface of rat BMSC is higher than 95%, and the positive rate of CD34 and CD45 is lower than 5%. The maximum non-toxic concentration of flavored salvia beverage to BMSC is 0.625g/L. Desmin, α-MHC and cTnT protein expression levels are higher than those of the normal control group, including Kawei Danfeng Decoction group, 5-azacytidine group, Kaweitan deep soup + 5-azacytidine group and GATA-4, in CX43 and Nkx2-5 mRNA is up-regulated. The expression level was up-regulated, and the difference was statistically significant (P\u003c0.01). Compared with Jiawei Danshen Decoction group or 5-azacytidine group, Jiawei Danshen Decoction + 5-azacytidine group, the expression of GATA-4, CX43 and Nkx2-5 mRNA all up-regulated Desmin, α-MHC and cTnT Protein expression. The level of expression is increased. The difference is statistically significant (P\u003c0.01).

　　Conclusion: Jiawei Danshen Decoction can induce bone marrow mesenchymal stem cells to transform into cardiomyocyte-like cells, and it has a synergistic effect when combined with 5-azacytidine.