Inhibitory effect of antisense oligonucleotide targeting TIMP-2 on immune-induced liver fibrosis

Dig Dis Sci. 2010 May;55(5):1286-95. doi: 10.1007/s10620-009-0858-5. Epub 2009 Jun 11.

Abstract

Introduction: We previously reported that both experimental and human studies have shown the importance of TIMP-1 and TIMP-2 in the development of liver fibrosis, a disease mostly caused by HBV and HCV infection in China. Inhibiting the expression of TIMP-1 by an antisense oligonucleotide (ASON) can prevent liver fibrosis through decreasing the deposition of collagen I and III. Whether blocking the expression of TIMP-2 has the same effect on liver fibrosis is not clear.

Materials and methods: To interfere with this potentially effective target, we designed and synthesized two different ASON targeting TIMP-2, then mixed and transfected them by hydrodynamic injection into the rat livers with immune-induced liver fibrosis. We isolated HSCs from the HSA-induced rat model with liver fibrosis, and transfected them with ASON or sense oligonucleotide in vitro.

Results: We observed that TIMP-2 ASON markedly reduced the expression of TIMP-2 by real-time PCR, Western blot, and enzyme linked immunosorbent assay. However, TIMP-2 ASON had little effect on alpha-SMA expression in vitro by Western blot. Inhibition of the expression of TIMP-2 by TIMP-2 ASON clearly decreased deposition of collagen I and IV, ameliorated liver pathology, and improved the liver function among the rats with immune-induced liver fibrosis.

Conclusion: The results suggested that TIMP-2 ASON could prevent the progression of liver fibrosis in this rat model. It is possible that this could form the basis for exploration of new liver anti-fibrosis drugs at a genetic level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Chromatography, High Pressure Liquid
  • Collagen / metabolism
  • Disease Progression
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Genetic Therapy / methods
  • Liver Cirrhosis / immunology
  • Liver Cirrhosis / prevention & control*
  • Oligonucleotides, Antisense / genetics
  • Oligonucleotides, Antisense / pharmacology*
  • Random Allocation
  • Rats
  • Rats, Wistar
  • Reverse Transcriptase Polymerase Chain Reaction
  • Serum Albumin
  • Statistics, Nonparametric
  • Tissue Inhibitor of Metalloproteinase-2 / genetics*

Substances

  • Oligonucleotides, Antisense
  • Serum Albumin
  • Tissue Inhibitor of Metalloproteinase-2
  • Collagen