动物造模,骨髓间充质干细胞 z-bio 2022-07-01 449

　　Objective: To investigate the experimental study on the differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells induced by Modified Danshen Decoction in vitro.

　　Methods: Rat BMSCs were obtained by the adherent method, and cell surface antigens were detected by flow cytometry. MTT method was used to detect the maximum non-toxic concentration of BMSCs with Jiawei Danshen Decoction, and the third-generation cells were used to group and induce: normal control group, Jiawei Danshen Decoction group, 5-azacytidine group, Jiawei Danshen Decoction + 5-azacytidine group, using western The expression of cell-binding protein (Desmin), α-myosin heavy chain (α-MHC) and cardiac troponin T (cTnT) in each group was detected by blot, and GATA-4 and connexin were detected by RT-PCR in each group. 43 (CX43) and Nkx2-5 mRNA expression.

　　RESULTS: The positive rate of CD90 on the surface of rat BMSCs was higher than 95%, and the positive rate of CD34 and CD45 was lower than 5%. The maximum nontoxic concentration of modified Danshen Decoction to BMSCs was 0.625 g/L. Compared with the normal control group, the expressions of Desmin, α-MHC and cTnT proteins, GATA-4, CX43 and Nkx2-5 were up-regulated in the Jiawei Danshen Yin group, the 5-azacytidine group, and the Jiawei Danshen Yin + 5-azacytidine group. mRNA expression was up-regulated, and the difference was statistically significant (P<0.01). Compared with the modified Danshen decoction group or the 5-azacytidine group, the protein expressions of Desmin, α-MHC and cTnT in the modified Danshen decoction + 5-azacytidine group were up-regulated, and the mRNA expressions of GATA-4, CX43 and Nkx2-5 were up-regulated , the difference was statistically significant (P<0.01).

　　Conclusion: Modified Danshen Decoction can induce the transformation of BMSCs into cardiomyocyte-like cells, and the combined administration with 5-azacytidine has a synergistic effect.