Effect of shRNA mediated Smad4 gene silencing on the fibrosis of C2C12 myoblasts

On tgf-β1 induced fibrosis. shrnas targeting smad4 were designed and the most efficient shrna was screened. this shrna was introduced into a lentiviral vector which was used to infect c2c12 myoblasts,and then the smad4 expression was detected. cells were divided into: c2c12 cells group,tgf-β1 induction group,transfection group,and transfection after tgf-β1 induction group. c2c12 myoblasts were transfected with lentivirus carrying smad4-shrna and treated with tgf-β1 to induce the differentiation into myofibroblasts. fluorescence real-time-pcr and the western blot assay were employed to detect the expressions of collagen i and α-sma. the results showed that the protein and mrna expression of smad4 in the c2c12 cells transfected with smad4-shrna1 was significantly reduced when compared with c2c12 before transfection. in the tgf-β1 induction group,the mrna expressions of α-sma and collagen i were significantly increased as compared to the c2c12 cells group. in the transfection after tgf-β1 induction group,the mrna expressions of α-sma and collagen i were significantly increased compared to the transfection group,and the protein expressions significantly increased,respectively. in the transfection after tgf-β1 induction group,the mrna expressions of α-sma and collagen i were significantly decreased compared to the tgf-β1 induction group,and the protein expressions significantly reduced,respectively. the results indicate that suppression of smad4 expression can efficiently inhibit the tgf-β1 induced fibrosis in myoblasts. the findings suggest smad4 may become a novel target for the treatment of skeletal muscle fibrosis. © journal of applied biomedicine.