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


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.


CONCLUSIONS
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.
Nie
Qing-He
QH
Chinese PLA Centre of Diagnosis and Treatment for Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xinsi Road, 710038 Xi'an, Shaanxi Province, People's Republic of China. nieqinghe@163.com
Zhu
Chuan-Long
CL
Zhang
Ya-Fei
YF
Yang
Jie
J
Zhang
Jiu-Cong
JC
Gao
Ren-Tao
RT
D000818:Animals; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000378:metabolism; D018450:Disease Progression; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D008103:Liver Cirrhosis / Q000276:immunology / Q000517:prevention & control*; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012709:Serum Albumin; D018709:Statistics, Nonparametric; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics*
D016376:Oligonucleotides, Antisense; D012709:Serum Albumin; D019716:Tissue Inhibitor of Metalloproteinase-2; D003094:Collagen