title: Inhibition of connective tissue growth factor suppresses hepatic stellate cell activation in vitro and prevents liver fibrosis in vivo.
abstract: Activation of hepatic stellate cells (HSC) represents a critical event in fibrosis, and connective tissue growth factor (CTGF) plays a profibrotic activity and a key factor in the pathogenesis of tissue fibrosis. The current study aimed to determine whether lentivirus-mediated short hairpin RNA (shRNA)-targeted CTGF downregulates the CTGF expression and furthermore whether it suppresses the activation and proliferation of HSC in vitro and prevents liver fibrosis in vivo. HSC-T6 cells were treated with recombinant lentivirus carrying CTGF siRNA. Real-time PCR, Western blotting, MTT, and flow cytometry were performed to investigate the activation and proliferation of HSC-T6 cells in response to CTGF silence. CCl4-induced rats were received lentivirus containing CTGF siRNA by intraportal vein injection. Levels of liver fibrosis were assessed by biochemical and histopathologic examinations. Recombinant lentivirus containing CTGF siRNA could effectively and specifically downregulate the expression of CTGF in both HSC-T6 cells and CCl4-induced rats with liver fibrosis. Blockade of CTGF resulted in significant inhibition of HSC activation and proliferation with decrease in TIMPs, MMP2, MMP9, and collagen I, as well as increase in cells in S phase. Silencing CTGF expression with siRNA prevented liver fibrosis in CCl4-induced rat model. These findings indicated that CTGF plays a key role in the pathogenesis of liver fibrosis and lentiviral-mediated CTGF siRNA has the potential to be an effective treatment for liver fibrosis.
authors: Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, 1 Xinsi Road, Xi'an, 710038, Shannxi Province, China.
authors: Chunqiu
authors: C
authors: Hao
authors: Yumei
authors: Y
authors: Xie
authors: Meijuan
authors: M
authors: Peng
authors: Li
authors: L
authors: Ma
authors: Yun
authors: Y
authors: Zhou
authors: Yan
authors: Y
authors: Zhang
authors: Wenzhen
authors: W
authors: Kang
authors: Jiuping
authors: J
authors: Wang
authors: Xuefan
authors: X
authors: Bai
authors: Pingzhong
authors: P
authors: Wang
authors: Zhansheng
authors: Z
authors: Jia
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D055513:Connective Tissue Growth Factor / Q000378:metabolism* / Q000627:therapeutic use*; D004195:Disease Models, Animal; D005434:Flow Cytometry; D020868:Gene Silencing; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000502:physiology*; D006651:Histocytochemistry; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008103:Liver Cirrhosis / Q000473:pathology / Q000517:prevention & control*; D008297:Male; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D013194:Staining and Labeling; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D013778:Tetrazolium Salts; D013844:Thiazoles; D055513:Connective Tissue Growth Factor; C022616:thiazolyl blue