[The construction of siRNA-tTG lentivirus vector and its effect on activated hepatic stellate cells].

OBJECTIVE

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).

METHODS

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 groups:negative 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.

RESULTS

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).

CONCLUSIONS

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.