The effect of Smad3 knockdown on the secretion of extracellular matrix induced by IGFBPrP1 in activated hepatic stellate cells / 中华肝胆外科杂志

ABSTRACT

Objective To identify the effect of IGFBPrP1 on the secretion of extracellular matrix in hepatic stellate cells through the Smad3 signaling pathway. Methods (1)Two pairs of chemically synthesized siRNAs (siRNA1, siRNA2) targeting Smad3 were transfected into HSC-T6 cells,real-time PCR and Western blot were used to evaluate the silence efficiency, and the better siRNA was used. (2)HSC-T6 cells were divided into four groups Negative control group, siRNA-Smad3 transfection group, siRNA-Smad3+IGFBPrP1 group and IGFBPrP1 group. The better siRNA was chosen to transfect into HSC-T6 cells. The protein expressions of Smad3, fibronectin and Collagen Ⅰ were evaluated by Western blot. Results (1)siRNA2-Smad3 inhibited Smad3 gene expression stronger than another siRNA. (2)After transfection of siRNA2-Smad3, the protein expression of Smad3 was significantly decreased compared to the negative control group(P＜0.01). The protein expression of fibronectin and Collagen Ⅰ in IGFBPrP1 stimulating HSCs treated with siRNA2-Smad3 were significantly decreased compared to that in IGFBPrP1 stimulating HSC without siRNA2-Smad3 (P ＜0. 01 ).Conclusion IGFBPrP1 induces the secretion of extracellular matrix in hepatic stellate cells through the Smad3 signaling pathway.