丹酚酸B体外诱导MSCs向心肌细胞分化中β-catenin表达的实验研究

Study on the β-catenin expression in the process of salvianolic acid B induced differetiation of MSCs into cardiomyocytes in vitro

关键词:  丹酚酸B  骨髓间充质干细胞  心肌细胞  Wnt信号

Key words:  Salvianolic acid B  MSCs  cardiomyocytes  Wnt signaling

摘要:

[目的] 体外诱导大鼠骨髓间充质干细胞(MSCs)向心肌细胞分化,检测分化过程中β-catenin蛋白表达的变化。[方法] 大鼠MSCs经采用密度梯度离心法和贴壁筛选法培养,免疫细胞化学法检测MSCs的表面抗原表达,取纯化稳定的P3代MSCs,将其分为4组:空白对照组、5-氮胞苷组、丹酚酸B组、5-氮胞苷联合丹酚酸B组、进行诱导,诱导4周,观察细胞形态学改变;免疫组化方法检测心肌特异性蛋白cTnT的表达,逆转录--聚合酶链反应 (RT-PCR)方法检测MSCs诱导前后糖原合成激酶(GSK)、β-catenin基因 mRNA表达。[结果] 诱导分化后的各组不同程度的阳性表达心肌细胞特异性蛋白心肌肌钙蛋白T(cTnT),RT-PCR 显示诱导组GSK基因表达上调,β-catenin基因表达下调。[结论] 丹酚酸B促进MSCs向心肌细胞分化,并伴有wnt经典信号通路的抑制。

Abstract:

[Objective] To discuss the role of salvianolic acid B in inducing differention of MSCs into cardiomyocytes and the β-catenin expression in the induced MSCs.[Methods] A method has been established of culturing MSCs from rat bone marrow. The third-passage MSCs were divided into four groups: the control group, the 5-Azacytidine group, the salvianolic acid B group, the 5-Azacytidine combined with salvianolic acid B group. After induced for 4 weeks, cardiac-specific cTnT expression was detected by immunocytochemistry and the mRNA expression of β-catenin and GSK was detected by RT-PCR in each group. [Results] The expression of cTnT in each induced group was significantly higher than that in the control group. The mRNA expression of GSK in each induced group was significantly up-regulated and that of β-catenin was down-regulated when compared with control group. [Conclusion] SalB can promote the differention of cardiomyocytes from MSCs and there is a typical inhibition of the Wnt signaling in this process.