The Molecular Mechanism And Experimental Study Of SFRP5-mediated Wnt Signaling Pathway In Preeclampsia

OBJECTIVE: Preeclampsia(PE)is a serious pregnancy complication.Abnormalities of extravillous trophoblast(EVT)invasion and spiral artery remodeling are the key to its pathogenesis.As a negative regulator of Wnt signaling pathway,the role of sFRP5 in the process of trophoblast invasion,angiogenesis and other processes is not yet clear.The purpose of this study was to detect the differences in the expression of sFRP5 in the serum of women with preeclampsia and normal pregnancy.To investigate the role and possible mechanisms of sFRP5 in the regulation of the invasion and migration of human trophoblasts and umbilical vein endothelial cells.To explore the role of sFRP5 involved in the possible molecular mechanisms of preeclampsia.METHODS: Fifteen normal pregnant women with full-term deliveries for cesarean section and fifteen preeclamptic women’s preoperative serum were selected at the Department of Obstetrics of the First Affiliated Hospital of Chongqing Medical University.Enzyme-linkedimmunosorbent assay(ELISA)techniques were used to detect the secretion of sFRP5 protein in the serum;Lentiviral vectors were used to increase sFRP5 gene expression in the extravillous trophoblastic cells HTR8/svneo and human umbilical vein endothelial cells HUVEC;flow cytometry,fluorescence microscopy,and western bloting were used to detect the efficiency of transfection;LiCl was used to activate the Wnt/β-catenin signaling pathway;cell proliferation was detected by CCK8assay;the invasion of HTR8/svneo cells was detected by transwell and Matrigel invasion assay;the migration of HUVEC cells was detected by scratch test.Western blotting was used to detect the effect of sFRP5 on the expression of β-catenin and other proteins.RESULTS: ELISA indicated that the expression of sFRP5 protein in the serum of preeclampsia(PE)group was significantly higher than that of the control group;the transfection efficiency of s FRP5 was over 95%;western bloting indicated the overexpression was effective,and the cck8 assay showed that Li Cl had a significant effect on the proliferation of the two cell lines for 24 h under the concentration of 20μM.The transwell and Matrigel invasion assay suggested that the invasiveness of HTR8/svneo cells was significantly reduced by sFRP5.After 24 hours’ treatment with Li Cl,the invasiveness of the cells in the overexpression group was rescued.The scratch test showed that the migration ability of HUVEC cells after theoverexpression was significantly decreased.After 24 h’ treatment with Li Cl.Western blotting detection revealed that the expression of β-catenin and p-gsk3β was decreased after overexpression of sFRP5.LiCl rescued the expression of β-catenin and p-gsk3β and the migration of the two cell lines.Conclusion: sFRP5 may impair the invasion and migration of trophoblast cells and endothelial cells through inhibiting the Wnt/β-cateninin signaling pathway,and which is involved in the occurrence and development of preeclampsia.