Mechanism Of EZHU-containing Serum Regulating The Expression Of Correlation Factor Of Hedgehog And Wnt Signaling Pathway In HSCs

ObjectiveTo explore the mechanism of Ezhu-containing serum regulating the expression of correlation factor of hedgehog and Wnt signaling pathway in hepatic stellate cells(HSCs)stimulated by leptin.MethodsThirty sprague-dawley(SD)rats were divided into 3 groups at random(n=10),ezhu-decoction group,colchicine group,physiological saline group.Rats in each group were intragstrically admmistered with the corresponding medicines,respectively,once perday for 10 days.Then,the drug-containing serums were collected and added to the cultured HSCs.The method of high performance liquid chromatography(HPLC)was used to detect the composition of Ezhu in serum.The HSCs during the exponential growth phase were divided into 8 groups: blank control group,model group,hedgehog pathway inhibitors(cyclopamine)group,Ezhu group,Ezhu and cyclopamine group,hedgehog pathway agonist(pumorphamine)group,Ezhu and purmorphamine group,colchicine group.HSCs were cultured invitro and stimulated with 0.1 μg/m L leptin(except for the blank control group),then were treated separately with the corresponding medicines for24 hours.After cells being collected,HSCs proliferation was detected using MTT colorimetric assay,the expression of Shh,Gli1,SFRP1 and β-catenin were determined by PT-PCR,Western-blot and immunofluorescence,respectively.ResultsWith HSCs stimulated by leptin,the expression of Shh,Gli1 andβ-catenin were significantly up-regulated than the blank control group,while the expression of SFRP1 were significantly down-regulated than the blank control group.After interfered by cyclopamine,Ezhu-containing serum and colchicines,the expression of Shh,Gli1,β-catenin were clearly down-regulated than the model group,while the expression of SFRP1 were clearly up-regulated than the model group.After concerted intervention with cyclopamine and Ezhu-containing serum,the expression of Shh,Gli1,β-catenin were significantly down-regulated than the model group,while the expression of SFRP1 were significantly up-regulated than the model group.After interfered by purmorphamine,the expression of Shh,Gli1,β-catenin were significantly up-regulated than the model group,while the expression of SFRP1 were significantly down-regulated than the model group.After using Ezhu-containing serum to interfere the purmorphamine group,the expression of Shh,Gli1,β-catenin significantlydown-regulated than the purmorphamine group,while the expression of SFRP1 significantly up-regulated than the purmorphamine group.ConclusionEzhu-containing serum plays an important part in inhibition of HSCs activation and increment.It may be achieved through regulating the expression of correlation factors of hedgehog and wnt signaling pathway.