The Protective Effect Of Taraxasterol On Liver Fibrosis And Its Mechanism Study

Objective: The purpose of this study was to investigate the effects of TAR in liver fibrosis and to reveal its possible mechanism by RNA sequencing.Methods: In vivo experiment,25% CCl4 was injected into the abdominal cavity of mice to establish liver fibrosis model.TAR was given to treat liver fibrosis.The effects of TAR on pathological morphology,biochemical indexes,oxidative stress,inflammatory response and ECM in mice with liver fibrosis were observed.The hepatoprotective mechanisms of TAR were analyzed by liver RNA sequencing.Real-time PCR and Western blot were used to verify the RNA sequencing results.In vitro experiment,TGF-β1 was used to establish liver fibrosis model.Results: TAR attenuated CCl4-induced hepatocyte necrosis,inflammatory infiltration and ECM deposition.TAR inhibited the levels of ALT,AST,ALP,γ-GT,LN,HA,PC III and IV-C in serum and TNF-α,IL-6,IL-1β and MDA in liver.In addition,TAR increased the activities of SOD and GSH-Px in liver.RNA sequencing analysis of liver tissues revealed that CCl4 and TAR significantly altered4,155 genes and 2,675 genes,respectively.TAR reversed changes in ECM-related genes.More specifically,TAR mediated the expression of genes related to the activation of the Hippo pathway,while inhibiting the expression of genes related to the activation of HIF-1α,TGF-β/Smad,and Wnt pathways.In the validation experiments,the q RT-PCR results showed that the expression levels of Yap1,Tead3,Hif1α,Vegfa,Tgfβ1,Want3 a,and Ctnnb1 m RNA were consistent with the RNA sequencing results.The Western blot results showed that TAR inhibited the levels of TGF-β1 and p-Smad2.In addition,the results in vitro were consistent with those in vivo.Conclusion: TAR improved CCl4-induced liver fibrosis by regulating Hippo,HIF-1α,TGF-β/Smad and Wnt pathways.