Get access

Inhibition of connective tissue growth factor by siRNA prevents liver fibrosis in rats

Authors

  • Guangming Li,

    1. Department of Gastroenterology, Xinhua Hospital, Shanghai Second Medical University, Shanghai 200092, China
    Search for more papers by this author
  • Qing Xie,

    1. Department of Infectious Diseases, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China
    Search for more papers by this author
  • Yi Shi,

    1. State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
    Search for more papers by this author
  • Dingguo Li,

    1. Department of Gastroenterology, Xinhua Hospital, Shanghai Second Medical University, Shanghai 200092, China
    Search for more papers by this author
  • Mingjun Zhang,

    1. Department of General Surgery, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China
    Search for more papers by this author
  • Shan Jiang,

    1. Department of Infectious Diseases, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China
    Search for more papers by this author
  • Huijuan Zhou,

    1. Department of Infectious Diseases, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China
    Search for more papers by this author
  • Hanming Lu,

    1. Department of Gastroenterology, Xinhua Hospital, Shanghai Second Medical University, Shanghai 200092, China
    Search for more papers by this author
  • Youxin Jin

    Corresponding author
    1. State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
    • State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue-yang Road, Shanghai 200031, China.
    Search for more papers by this author

Abstract

Background

Connective tissue growth factor (CTGF) is a highly profibrogenic molecule implicated in hepatic fibrogenesis. Small interfering RNA (siRNA) is an effective tool to silence gene expression post-transcriptionally. Therefore, we conducted an investigation to determine if intraportal vein siRNA injection targeting CTGF inhibits CTGF expression on rat liver in vivo and furthermore whether it protects the liver from liver fibrosis.

Methods

Some rats received carbon tetrachloride (CCl4) by subcutaneous injections every three days for six consecutive weeks, and meantime they also obtained either siRNA (0.1 mg/kg) targeting CTGF, saline or a control siRNA by intraportal vein injection to rats' liver at the same pattern. Other rats received CCl4 by subcutaneous injection for 2 weeks, followed by CCl4 and CTGF siRNA intraportal vein injection for four more weeks.

Results

Intraportal vein injection of CTGF siRNA specifically reduced the expression of CTGF protein in rat liver, and these effects were maintained for 3 days. Six weeks after CCl4 injection, prominent upregulations were observed in the gene expressions of CTGF, type I, III collagen, laminin, tissue inhibitor metal proteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in saline or control siRNA-treated rats livers. Administrating CTGF siRNA for 4 or 6 weeks, by contrast, markedly attenuated the induction of CTGF, type I, III collagen, laminin, TIMP-1 and TGF-β1 genes, whereas Smad2, 7 gene expression was not affected. The number of active hepatic stellate cells (HSCs) determined by the expression of α-smooth muscle actin was also significantly decreased. The CTGF siRNA treatment markedly reduced serum procollagen type III, hepatic hydroxyproline and liver fibrosis staging.

Conclusions

Silencing CTGF expression with siRNA demonstrates therapeutic potential to prevent liver fibrosis by inhibiting HSC activation with consequent extracellular matrix accumulation and the upregulation of TGF-β1 gene expression. Copyright © 2006 John Wiley & Sons, Ltd.

Ancillary