动物造模,骨髓间充质干细胞 z-bio 2023-05-08 583

　　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 nontoxic concentration of Jiawei Danshen Decoction on BMSCs, and the third-generation cells were used to induce grouping: 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.