Baocan Wang, Wenxi Li, Yingwei Chen, Yuqin Wang, Chao Sun, Yuanwen Chen, Hanming Lu, Jiangao Fan, Dingguo Li
Med Sci Monit 2012; 18(10): BR394-401
Available online: 2012-09-28
Background: There is a great need for developing novel therapies to treat liver fibrosis. Previous studies showed that both Smad7 and uPA were inhibitors of liver fibrosis. Therefore, we explored the therapeutic effects of combinational gene therapy with Smad7 and uPA on CCl4-induced liver fibrosis.
Material/Methods: Smad7 and uPA genes were cloned into an adenovirus vector. To observe the therapeutic effects of coexpression of Smad7 and uPA genes, the recombinant adenovirus were delivered into CCL4-induced fibrosis models. Fibrillar collagen, hydroxyproline, α-SMA, TGF-β1, MMP-13, TIMP-1, HGF and PCNA were detected to evaluate the fibrosis and to explore the mechanisms underlying the treatment with Smad7 and uPA.
Results: The results showed that single Smad7 or uPA adenovirus reduced CCL4 induced liver fibrosis significantly; while combination of Smad7 and uPA had more significant therapeutic effect on CCl4 induced liver fibrosis. Then the markers underlying the therapeutic effect of combination of Smad7 and uPA were also explored. Over-expression of Smad7 and uPA inhibited the expression of α-SMA and TGF- β1 significantly. Combinational gene therapy also enhanced extracellular matrix degradation by increasing the expression of MMP-13, inhibiting TIMP-1 expression, and promoted hepatocyte proliferation, while single Smad7 or uPA only induced part of these changes.
Conclusions: These results suggest that combinational gene therapy with Smad7 and uPA inhibited CCl4-induced rat liver fibrosis by simultaneously targeting multiple pathogenic pathways.
Keywords: Recombination, Genetic - genetics, Rats, Rats, Sprague-Dawley, RNA, Messenger - metabolism, Matrix Metalloproteinase 13 - metabolism, Liver Cirrhosis - therapy, Liver - pathology, Hepatocytes - pathology, genetic therapy, Gene Expression Regulation, Enzymologic, Extracellular Matrix - metabolism, Disease Progression, Cell Proliferation, Animals, Carbon Tetrachloride, Adenoviridae - genetics, Smad7 Protein - therapeutic use, Tissue Inhibitor of Metalloproteinase-1 - metabolism, Transforming Growth Factor beta1 - metabolism, Urokinase-Type Plasminogen Activator - therapeutic use