丹参抑制转化生长因子-β1诱导的肌源性干细胞向肌成纤维母细胞分化

Inhibitory effect of Salvia miltiorrhiza on myofibroblastic differentiation of skeletal muscle-derived stem cells induced by transformation growth factor-β1

目的观察丹参对失神经骨骼肌肌源性干细胞（MDSCs）向肌成纤维母细胞分化的抑制作用。方法采用差速贴壁法分离出大鼠失神经骨骼肌MDSCs，加入转化生长因子-β1（TGF-β1）和丹参进行干预，将细胞分为3组：A：对照组；B：10μg／LTGF-β1组；C：10μg／LTGF-β1＋150mg／L丹参组。荧光实时定量聚合酶链反应（qRT—PCR）和Westernblot检测各组细胞在干预后5个时间点α-平滑肌肌动蛋白（α-SMA）和波形蛋白（Vimentin）mRNA和蛋白表达。结果与A组比较，B组和C组细胞α-SMA、Vimentin的mRNA在干预后第2、3、5、7天均明显升高（P〈0．05），其蛋白表达在干预后第3、4、6、8天亦显著升高（P〈0．05）；与B组比较，C组细胞α-SMA、Vimentin的mRNA和蛋白在对应时间点均明显降低（P〈0．05）。结论丹参能抑制TGF-β1诱导的失神经骨骼肌MDSCs向肌成纤维母细胞的分化。

Objective To investigate the inhibitory effect of Salvia miltiorrhiza on differentiation of skeletal muscle-derived stem cells into myofibroblasts. Methods MDSCs in SD rats were isolated by differential adhesion method. Differentiation of MDSCs was induced by transformation growth factor-β1 （TGF-β1） administration. The cells cultivated in vitro was divided into three groups ： control group （ A ） , 10μg/L TGF-β1 group （B） , 10 μg/L TGF-β1 ＋ 150 mg/L Salvia miltiorrhiza group （C）. The mRNA expression levels of α-smooth muscle actin （α-SMA） and Vimentin in each group were detected by quantiteve realtime reverse transcription-polymerase chain reaction （qRT-PCR） and there protein secretions were detected by Western blotting. Results As compared with group A, the mRNA after TGF-β1 intervention and protein expression of α-SMA and Vimentin at the 3rd, 4nd, 6nd, 8nd day in groups B and C were prominently increased （P 〈 0. 05 ）. As Compared with group B, the mRNA and protein expression levels of α-SMA and Vimentin in group C were obviously down-regulated （P 〈 0. 05 ）. Conclusion Salvia mihiorrhiza could inhibit myofibroblastic differentiation and further ameliorate fibrosis of denervated skeletal muscles.