丹参抑制骨骼肌肌源性干细胞成纤维样分化作用的研究

目的 研究丹参对体外培养的骨骼肌肌源性干细胞(MDSCs)纤维样分化的作用.方法 采用差速贴壁法分离出大鼠MDSCs,利用TGF-β1诱导MDSC向成纤维样细胞分化,观察丹参对细胞成纤维样分化的影响.将体外培养的细胞分为3组:A组,对照组;B组,10 ng/ml TGF-β1组;C组,10 ng/ml TGF-β1+150 mg/L丹参组.在放大400倍相差显微镜下观察细胞生长情况,利用免疫细胞化学方法、RT-PCR、Western Blot检测MDSCs表达CTGF、COLⅠ、COL Ⅲ的情况.结果 在体外,TGF-β1能加强MDSCs表达结缔组织生长因子(CTGF)和胶原蛋白,丹参拮抗TGF-β1的作用,降低细胞CTGF、COLⅠ、COL Ⅲ mRNA的表达水平,降低CTGF、COLⅠ、COL Ⅲ蛋白的水平.结论 丹参可以抑制TGF-β1诱导MDSCs成纤维样分化作用.其机制可能是抑制CTGF表达,从而降低细胞合成和分泌胶原纤维,抑制细胞成纤维样分化.

Abstract：

Objective To investigate the effect of Salvia miltiorrhiza on fibroblastic differentiation of skeletal muscle-derived stem cells (MDSCs) in vitro. Methods MDSCs in rats were isolated by the method of adhering to culture plastic at different time points. Fibroblastic differentiation of MDSCs was induced by TGF-β1 administration and the effects of Salvia miltiorrhiza on fibroblastic differentiation were observed. The cells cultivated in vitro were divided into three groups according the supplement of TGF-β1 and Salvia miltiorrhiza in culture media: group A (control group) had no TGF-β1 and Salvia miltiorrhiza;group B had 10 ng/ml TGF-β1;group C had 10 ng/ml TGF-β1 and 150 mg/L Salvia miltiorrhiza. Cell growth was observed using phase contrast microscope. The expressions of CTGF,COLⅠand COL Ⅲ in each group were detected by immunocytochemistry,RT-PCR and Western Blot. Results In vitro,TGF-β1 promoted the expression of CTGF and collagens of MDSCs. However,the effect of TGF-β1 could be inhibited by Salvia miltiorrhiza,resulting in decreased mRNA and protein expression of CTGF,COLⅠand COL Ⅲ. Conclusion Salvia miltiorrhiza could inhibit fibroblastic differentiation of MDSCs induced by TGF-β1. The mechanism might be that inhibition of CTGF expression reduces the synthesis and secretion of collagen fibers and in turn inhibits the fibroblastic differentiation.

Inhibitory effect of Salvia miltiorrhiza on fibroblastic differentiation of skeletal muscle-derived stem cells

Objective To investigate the effect of Salvia miltiorrhiza on fibroblastic differentiation of skeletal muscle-derived stem cells (MDSCs) in vitro. Methods MDSCs in rats were isolated by the method of adhering to culture plastic at different time points. Fibroblastic differentiation of MDSCs was induced by TGF-β1 administration and the effects of Salvia miltiorrhiza on fibroblastic differentiation were observed. The cells cultivated in vitro were divided into three groups according the supplement of TGF-β1 and Salvia miltiorrhiza in culture media: group A (control group) had no TGF-β1 and Salvia miltiorrhiza;group B had 10 ng/ml TGF-β1;group C had 10 ng/ml TGF-β1 and 150 mg/L Salvia miltiorrhiza. Cell growth was observed using phase contrast microscope. The expressions of CTGF,COLⅠand COL Ⅲ in each group were detected by immunocytochemistry,RT-PCR and Western Blot. Results In vitro,TGF-β1 promoted the expression of CTGF and collagens of MDSCs. However,the effect of TGF-β1 could be inhibited by Salvia miltiorrhiza,resulting in decreased mRNA and protein expression of CTGF,COLⅠand COL Ⅲ. Conclusion Salvia miltiorrhiza could inhibit fibroblastic differentiation of MDSCs induced by TGF-β1. The mechanism might be that inhibition of CTGF expression reduces the synthesis and secretion of collagen fibers and in turn inhibits the fibroblastic differentiation.