Ribozymes against TGFβ1 reverse character of activated hepatic stellate cells in vitro and inhibit liver fibrosis in rats
✍ Scribed by Yu-Hu Song; Xiang-Long Chen; Xin-Juan Kong; Nan-Zhi Liu; Wei Li; Xiao-Li Wu; Ju-Sheng Lin; You-Xin Jin
- Publisher
- John Wiley and Sons
- Year
- 2005
- Tongue
- English
- Weight
- 362 KB
- Volume
- 7
- Category
- Article
- ISSN
- 1099-498X
- DOI
- 10.1002/jgm.744
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✦ Synopsis
Abstract
Background/aims
Transforming growth factor β (TGFβ1) is considered the key mediator in the process of liver fibrosis. The purpose of this investigation was to evaluate the activity of ribozymes against TGFβ1 in a cell‐free system and activated hepatic stellate cells (HSCs), and antifibrotic effect in activated HSCs in vitro and in rats.
Methods
Three ribozymes targeting against TGFβ1 mRNA were designed, and then cloned into the U1 snRNA expression cassette. The chimeric ribozymes were selected for the analysis of their performances in activated HSCs through the detection of their cleavage activities in a cell‐free system. After ribozyme‐encoding plasmids had been transfected into HSC‐T6 cells, the effects of ribozymes on activated HSCs were evaluated through the analysis of proliferation, activation and collagen deposition of HSC‐T6. The adenoviral vector expressing the ribozymes was constructed, and then delivered into rat models of hepatic fibrosis induced by carbon tetrachloride.
Results
TGFβ1 expression was efficiently down‐regulated in activated HSCs by U1 snRNA chimeric ribozymes which possessed perfect cleavage activity in a cell‐free system. Further studies demonstrated that U1 snRNA chimeric ribozymes inhibited the synthesis of collagen I, reduced deposition of collagen I, suppressed BrdU incorporation, but had no effect on desmin and α‐SMA expression in transfected HSC‐T6 cells. Histological analysis demonstrated that the adenoviral vector expressing ribozyme (Rz803) could alleviate fibrotic pathology in rats treated with carbon tetrachloride.
Conclusions
The anti‐TGFβ1 ribozymes could reverse the character of activated HSCs in vitro and improve fibrotic pathology in vivo. It indicated that TGFβ1 could be considered as a novel candidate for a therapeutic agent against hepatic fibrosis. Copyright © 2005 John Wiley & Sons, Ltd.