The construction of siRNA-tTG lentivirus vector and its effect on activated hepatic stellate cells / 中华外科杂志

ABSTRACT

<p><b>OBJECTIVE</b>To construct the lentivirus vector of siRNA-tTG and investigate the effects and mechanism of lentivirus mediated tTG RNA interference (RNAi) on the hepatic stellate cell (HSC).</p><p><b>METHODS</b>The effective RNAi sequences targeted the tTG gene were designed and Lenti-siRNA-tTG was constructed by plasmid transfection. The rats were distributed into three groupsnegative control (CON) group, siRNA-GFP (GFP) group and siRNA-tTG (rtTG) group. After the infection of lentivirus mediated siRNA-tTG, the mRNA and protein expression of tTG, alpha-SMA and collagen-1, and the cell proliferative status of HSC were observed in the three groups respectively.</p><p><b>RESULTS</b>The lentivirus vector of siRNA-tTG was constructed successfully. After the infection of lentivirus mediated siRNA-tTG, the mRNA expression of tTG and collagen-1 was decreased significantly (P < 0.05), and the protein expression of tTG, lysine and collagen-1 was also decreased significantly (P < 0.01), in the same time the proliferative ability of HSC was decreased significantly (P < 0.05).</p><p><b>CONCLUSIONS</b>Lentivirus mediated siRNA-tTG can inhibit the proliferation of HSC by inhibition of the expression of tTG and attenuate the synthesis of collagen-1 and lysine. Thus, it may have a potential anti-fibrosis effect.</p>