动物造模,骨髓间充质干细胞 z-bio 2021-01-28 304

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

　　Method: Obtain rat BMSCs by adhesion 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 cells (normal control group, Jiawei Danshen Decoction group, 5-azacitidine group, Jiawei Danshen Decoction + 5-azacitidine). Western blot detection group, cell binding protein (desmin), α-myosin heavy chain (α-MHC), cardiac troponin T (cTnT) protein expression in each group, RT-PCR method to detect GATA-4, gap expression Connexin 43 (CX43) and Nkx2-5 mRNA.

　　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 BMSC flavored salvia beverage is 0.625g/L. Compared with the normal control group, the expressions of GATA-4, CX43 desmin, α-MHC and cTnT protein were up-regulated in the Jiawei Danshen Decoction, 5-azacitidine group and Jiawei Danshen Decoction + 5-azacitidine group. And the expression level of Nkx2-5 mRNA was up-regulated, and the difference was statistically significant (P\u003c0.01). Compared with the Jiawei Danshen Decoction group or the 5-azacitidine group, the Jiawei Danshen Decoction + 5-azacitidine group and the expression levels of GATA-4 and CX43 increased the expression of Desmin, α-MHC and cTnT protein. kx2-5 mRNA has been upregulated. The difference is statistically significant (P\u003c0.01).

　　 Conclusion: Jiawei Danshen Decoction can transform bone marrow mesenchymal stem cells into cardiomyocyte-like cells, which has a synergistic effect with 5-azacitidine.