Secreted Frizzled-related Protein 2 Inhibits Hepatic Stellate Cells Activation And Proliferation In Liver Fibrosis

BACKGROUND: Liver fibrosis is a persistent pathological change caused by a chronic liver injury by various etiologies.The main manifestation of liver fibrosis is the excessive accumulation of extracellular matrix(ECM)in the liver.Liver fibrosis is the development of chronic liver disease to the liver.There is a close relationship between the proliferation and activation of hepatic stellate cells and the abnormal accumulation of extracellular matrix in hepatocytes during hepatic fibrosis.Meanwhile,a large number of reports have been reported that Wnt signaling pathway plays an important role in the proliferation and activation of hepatic stellate cells.Secreted frizzled related protein 2(SFRP2),a member of the secreted frizzled-related protein(SFRP)family,which acts as an antagonist to bind Wnt/?-catenin proteins to inhibit signal activation in many tumors.It plays an important role in cell proliferation and apoptosis.At the same time,this antagonism is mostly related to its own gene methylation.Because of the important role of ?-catenin in the activation and proliferation of hepatic stellate cells(HSCs),the functional regulation of SFRP2 on the proliferation and activation of hepatic stellate cells and related mechanisms were explored.METHODS: In vivo,the C57/BL6 mice were injected subcutaneously with carbon tetrachloride(CCl4)for 4 weeks and 2 times per week to establish a hepatic fibrosis model.The liver tissue and primary hepatic stellate cells in normal and model mice were taken to observe the liver pathological changes and detect the relevant indexes of hepatic fibrosis to confirm the successful establishment of hepatic fibrosis model.The expression of SFRP2 and DNA methylation-related proteins in mouse hepatic stellate cells was detected by Western Blot,RT-q PCR andimmunohistochemistry.In vitro,TGF-? activated HSC-T6,Western Blot and RT-q PCR method to detect SFRP2 expression changes.Cells transfected with DNMT1-si RNA,DNMT3a-si RNA,DNMT3b-si RNA and PEGFP-N1-SFRP2 were used to detect the expression of related proteins and cell proliferation by Western Blot and flow cytometry.Simultaneous detection of Wnt/?-catenin signaling pathway related protein expression changes to determine the relationship between SFRP2 and Wnt/?-catenin signaling pathways.5-Aza-d C(5-Aza-2'-deoxycytidine)is a DNA methylation inhibitor,and 5-Aza-d C-treated rat hepatic stellate cells were observed using Western Blot.RESULTS: In vivo experiments,the results showed that SFRP2 expression was significantly reduced in hepatic stellate cells of liver fibrosis mice.In vitro,the expression of SFRP2 was decreased in activated HSC-T6 cells;SFRP2 was overexpressed,the expression of liver fibrosis markers Col?1 and ?-SMA was decreased,and the expression of Wnt/?-catenin signaling pathway related proteins Cyclin D1,C-myc and ?-catenin were both decreased.Decreased;Silenced DNMT1,DNMT3 a,and DNMT3 b,decreased expression of Col?1 and ?-SMA,decreased expression of Cyclin D1,C-myc,and ?-catenin.CONCLUSION: In the process of hepatic fibrosis,the expression of SFRP2 in hepatic stellate cells is decreased,which promotes the proliferation and activation of hepatic stellate cells.This article suggests that SFRP2 may affect hepatic fibrosis by regulating the Wnt/?-catenin signaling pathway through its DNA methylation,leading to activation and proliferation of hepatic stellate cells.