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

Zhonghua Wai Ke Za Zhi. 2009 May 1;47(9):701-4.
[Article in Chinese]

Abstract

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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Genetic Vectors*
  • Hepatic Stellate Cells*
  • Lentivirus
  • RNA, Small Interfering / genetics*
  • Rats
  • Transfection
  • Transglutaminases / genetics*

Substances

  • RNA, Small Interfering
  • Transglutaminases