The antifibrotic effects of TGF-β1 siRNA on hepatic fibrosis in rats

• Published in: Biochemical and biophysical research communications 2011-06, Vol.409 (3), p.448-453
• Main Authors: Lang, Qing, Liu, Qi, Xu, Ning, Qian, Ke-Li, Qi, Jing-Hu, Sun, Yin-Chun, Xiao, Lang, Shi, Xiao-Feng
• Format: Article
• Language: English
• Subjects: Animals; Collagen Type I - genetics; Collagen Type I - metabolism; Collagen Type III - genetics; Collagen Type III - metabolism; Extracellular matrix; Gene therapy; Hepatic fibrosis; Hepatic stellate cells; Liver Cirrhosis - pathology; Liver Cirrhosis - therapy; Male; Plasmids - genetics; Rats; Rats, Sprague-Dawley; RNA Interference; RNA, Small Interfering - genetics; Short interference RNA; Transforming Growth Factor beta - antagonists & inhibitors; Transforming Growth Factor beta - genetics; Transforming Growth Factor beta - metabolism; Transforming growth factor-β1
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2011.05.023

► We constructed CCL4 induced liver fibrosis model successfully. ► We proofed that the TGF-β1 siRNA had a definite therapy effect to CCL4 induced liver fibrosis. ► The therapy effect of TGF-β1 siRNA had dose-dependent. Background/aims: Hepatic fibrosis results from the excessive secretion of matrix proteins by hepatic stellate cells (HSCs), which proliferate during fibrotic liver injury. Transforming growth factor (TGF)-β1 is the dominant stimulus for extracellular matrix (ECM) production by stellate cells. Our study was designed to investigate the antifibrotic effects of using short interference RNA (siRNA) to target TGF-β1 in hepatic fibrosis and its mechanism in rats exposed to a high-fat diet and carbon tetrachloride (CCL4). Methods: A total of 40 healthy, male SD (Sprague–Dawley) rats were randomly divided into five even groups containing of eight rats each: normal group, model group, TGF-β1 siRNA 0.125mg/kg treatment group, TGF-β1 siRNA 0.25mg/kg treatment group and TGF-β1 siRNA negative control group (0.25mg/kg). CCL4 and a high-fat diet were used for 8weeks to induce hepatic fibrosis. All the rats were then sacrificed to collect liver tissue samples. A portion of the liver samples were soaked in formalin for Hematoxylin–Eosin staining, classifying the degree of liver fibrosis, and detecting the expression of type I and III collagen and TGF-β1; the remaining liver samples were stored in liquid nitrogen to be used for detecting TGF-β1 by Western blotting and for measuring the mRNA expression of type I and III collagen and TGF-β1 by quantitative real-time polymerase chain reaction. Results: Comparing the TGF-β1 siRNA 0.25mg/kg treatment group to the model group, the TGF-β1 siRNA negative control group and the TGF-β1 siRNA 0.125mg/kg treatment group showed significantly reduced levels of pathological changes, protein expression and the mRNA expression of TGF-β1, type I collagen and type III collagen (P