加味丹参饮体外诱导骨髓间充质干细胞向心肌样细胞分化的实验研究
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Experimental study of Jiawei Danshen Yin on the differentiation of bone marrow mesenchymal stem cells into myocardium-like cells in vitro
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    摘要:

    目的 探讨加味丹参饮体外诱导骨髓间充质干细胞(BMSCs)向心肌样细胞分化的实验研究。方法 采用贴壁法获取大鼠BMSCs,流式细胞术检测细胞表面抗原。MTT法检测加味丹参饮对BMSCs最大无毒浓度,用第3代细胞分组诱导:正常对照组,加味丹参饮组,5-氮胞苷组,加味丹参饮 5-氮胞苷组,应用western blot检测各组细胞结合蛋白(Desmin),α-肌球蛋白重链(α-MHC),心肌肌钙蛋白T(cTnT)蛋白表达,RT-PCR法检测各组细胞GATA-4,缝隙连接蛋白43(CX43)及Nkx2-5 mRNA表达。结果 大鼠BMSCs表面CD90阳性率高于95%,CD34及CD45阳性率低于5%。加味丹参饮对BMSCs最大无毒浓度为0.625 g/L。与正常对照组比较,加味丹参饮组,5-氮胞苷组,加味丹参饮 5-氮胞苷组中Desmin,α-MHC及cTnT蛋白表达量上调,GATA-4,CX43及Nkx2-5 mRNA表达量上调,差异均具有统计学意义(P<0.01)。与加味丹参饮组或5-氮胞苷组比较,加味丹参饮 5-氮胞苷组中Desmin,α-MHC及cTnT蛋白表达量上调,GATA-4,CX43及Nkx2-5 mRNA表达量上调,差异均具有统计学意义(P<0.01)。结论 加味丹参饮能够诱导BMSCs向心肌样细胞转化,与5-氮胞苷联合给药具有协同作用。

    Abstract:

    Obejective To explore experimental study of Jiawei Danshen Yin on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into myocardium-like cells in vitro. Methods Rat BMSCs were isolated by adherent method, and their surface antigen were detected by flow cytometry. The maximal non-toxic concentration of Jiawei Danshen Yin was measured by MTT method. The isolated and purified BMSCs were divided into four groups: normal control group, Jiawei Danshen Yin group, 5-azacitidine group, Jiawei Danshen Yin plus 5-azacitidine group. The expression of desmin, α-myosin heavy chain (α-MHC) and cardiac troponin T (cTnT) protein was detected by western blot. The expression of GATA-4, gap junction connexin 43 (CX43) and Nkx2-5 mRNA was detected by RT-PCR. Results The positive rate of CD90 in the rat BMSCs was higher than 95%, and the positive rate of CD34 and CD45 was lower than 5%. The maximum toxic concentration of Jiawei Danshen Yin on BMSCs was 0.625 g/L. Compared with the normal control group, the expression of desmin, α-MHC and cardiac cTnT protein was up-regulated, the expression of GATA-4, CX43 and Nkx2-5 mRNA was up-regulated in Jiawei Danshen Yin group, 5-azacitidine group and Jiawei Danshen Yin plus 5-azacitidine group. The difference was statistically significant (P<0.01). Compared with Danshen Yin group or 5-azacitidine group, the expression of desmin, α-MHC and cardiac cTnT protein was up-regulated, the expression of GATA-4, CX43 and Nkx2-5 mRNA was up-regulated in Jiawei Danshen Yin plus 5-azacitidine group. The difference was statistically significant (P<0.01). Conclusion Jiawei Danshen Yin could induce BMSCs differentiation into myocardium-like cells, which had a synergistic effect with 5-azacitidine.

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韩析言.加味丹参饮体外诱导骨髓间充质干细胞向心肌样细胞分化的实验研究[J].中国比较医学杂志,,().

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  • 收稿日期:2016-04-26
  • 最后修改日期:2016-05-23
  • 录用日期:2016-05-23
  • 在线发布日期: 2016-11-28
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