Effects of siRNA targeting ADAMTS2 gene on hepatic stellate cells / 中华肝胆外科杂志

ABSTRACT

Objective To explore the preventive and therapeutic role of silencing type Ⅰ rat platelet-binding protein motifs depolymerization protein-like metalloproteinase 2(ADAMTS2)by siRNA on experimental liver fibrosis in vitro.By studying the mechanism of siRNA silencing of ADAMTS2,we also aim to evaluate the feasibility of ADAMTS2 as a target for anti-liver fibrosis therapy.Methods Three pairs of siRNAs targeting ADAMTS2 mRNA 2237,2597 and 690 targets were designed and synthesized by utilizing RNA design software.The most effective siRNA was chosen to transfect HSC-T6 cell line to test the tendency of hepatic stellate cell(HSC)activation and ex pression of ADAMTS2,COL1α1,COL(I),α-SMA,TGF-β1,MMP-2 and TIMP-3.These were quantified using real time-PCR,Western blotting,and MTT assays.Results Of the same dosage and time of injection,siRNA 2237 inhibited ADAMTS2 gene expression significantly more than other siRNAs.siRNA-ADAMTS2 2237 markedly inhibited ADAMTS2 gene and protein expression of HSCT6 with more than 80％ efficiency.Conversely,siRNA-ADAMTS2 2237 markedly reduced the gene and protein expressions of COL(I),α-SMA and TGF-β1 on HSC-T6 and inhibited the proliferation of HSC.Conclusions siRNA-ADAMTS2 2237 could effectively knockdown the gene and protein expression of ADAMTS2 in HSC-T6 cell lines.Silencing ADAMTS2 by siRNA significantly inhibited the activation,proliferation of HSC and the gene and protein expressions of COL(I),α SMA,and TGF-β1,and it may have a potential anti-fibrotic effect.ADAMTS2 might be an efficient target for anti-fibrotic therapy.