Compound A, An Inhibitor Of TGF-β Signaling, Suppresses Experimental Hepatic Fibrosis In Rats

Objective: Basing on the model of rats hepatic fibrosis induced by carbon tetrachloride (CCl₄) to investigate the effects of Compound A suppression on Hepatic Fibrosis in Rats.At the same time,construct a EGFP expression plasmid to response Wnt signaling.Methods: 1.Check the effects of compound A inhibites the TGF-β,BMP and Wnt signaling by Reporter Assay.2.Rats hepatic fibrosis assay.Throughout the study male Wistar rats weighing～250g were used.Animals were randomly divided into three groups including a healthy control group(n=6),a model control group(n=12) and a Compound A treatment group(n=12). Except group A, all groups were injected intraperitoneally (ip) with 50ï¼…CCl₄(2ml/kg/2day) for two weeks,to induce rat liver fibrosis.The treatment group were administrated with Compound A(3mg/kg/day).All rats were sacrificed after two weeks,the blood and livers of rats were collected for further examinations.The effects of Compound A were evaluated by detections of the mRNA level of TGF-β,Collagen typeâ… ,Collagen typeâ…¢,Fibronection,PAI-1,TGF-βand Smad7 by semi-quantitative RT-PCR,The level of Glutamate-pyruvate transaminase(ALT) and glutamic-oxalo acetic transaminase(AST) detected by spectrophotometer following the protocol of the Kit.Liver histopathology were also examined by HE stain and Mession stain.3.Basing on the two plasmids of CAGA-EGFP and SupetTopflash-luc,the EGFP expression plasmid responsing Wnt signaling was constructed.Results: 1.Compound A can suppresses TGF-βsignaling but can't influence BMP and Wnt signal pathway.2.Results in experimental animals, Compared with model group, in rats that received Compound A, the mRNA level of TGF-β, Fibronection,PAI-1,TGF-β,Smad7 and Collagen typeâ… ,Collagen typeâ…¢in liver tissues were significantly decreased (P＜0.05 or P＜0.01), the levels of serum ALT and AST were diminished (P＜0.1).3.The SuperTop8×Flash-EGFP plasmid well expression in 293Tcell and responses Wnt signal pathway.Conclusion: 1.Compound A suppresses TGF-βsignal pathway very well.2.Compound A can improve living status and suppress liver fibrosis.3.There are consanguineous relationship between Compound A and hepatic fibrosis.4.The SuperTop8×Flash-EGFP plasmid was constructed. Successfully.