Study Of RPS4Y1 Regulating HUVECs Functions And Its Role In The Pathogenesis Of Preeclampsia

OBJECTIVE: Preeclampsia(PE)is a unique and multisystem disease that occurs during pregnancy.It occurs 20 weeks after pregnancy and poses a great threat to the lives of pregnant women and perinatal mothers.The main manifestations of PE are new proteinuria and hypertension,and its incidence is reported to be about 3-5%.At present,the exact pathogenesis of preeclampsia is not yet clear,but studies have shown that the occurrence of PE is associated with placental ischemia-reperfusion injury,shallow placental implantation,vascular endothelial cell injury and immune imbalance.As a minor histocompatibility antigen,RPS4Y1 has been studied and confirmed that RPS4Y1 protein is expressed in all trophoblast cells and endothelial cells,and is mainly located in the cytoplasm.There is no relevant research on the function of RPS4Y1 in PE at home and abroad.Our research group aims to:(1)detect the localization and expression of RPS4Y1 in villi,decidua,term placenta,and placenta of PE;and(2)explore RPS4Y1 in human umbilical vein endothelial cell lines(HUVECs).Expression and its role and effects on the biological function of the cell;(3)HUVEC cells treated with hypoxia/reoxygenation to simulate oxidative stress injury in placenta tissue of preeclampsia combined with the JAK2 specific inhibitor AG490 to explore RPS4Y1 The regulation of the biological function of endothelial cells by JAK2/STAT3 signaling pathway and the possible molecular mechanism in the pathogenesis of PE provide theoretical basis and new ideas for the etiology and diagnosis of preeclampsia.Methods:(1)The expression and localization of RPS4Y1 protein in placenta tissues of villi,decidua,full-term placenta,and PE were detected by immunohistochemistry.(2)Real-time fluorescent quantitative PCR and WB were used to study the different expression of RPS4Y1 protein in placenta of patients with term placenta and severe PE.(3)RPS4Y1 gene expression in HUVEC cells was interfered by transfection with lentivirus.(4)Real-time fluorescence quantitative PCR and WB detection interference efficiency.(5)The effects of interference with RPS4Y1 expression on migration,angioplasty,proliferation,and apoptosis of HUVECs were examined by cell migration assay,lumen formation assay,CCK-8 assay,and flow cytometry assay,respectively.(6)Establishment of hypoxia/reoxygenation model in vitro,simulation of PE oxidative stress injury,detection of H/R injury on the biological function of HUVEC cells and regulation of RPS4Y1 expression.(7)The JAK2-specific inhibitor AG490 was used to observe the biological function of JAK2/STAT3 signaling pathway under H/R conditions on HUVEC cells,further clarify the role of RPS4Y1 gene in regulating the function of endothelial cells in the pathogenesis of PE and its possible mechanism.Results: 1.Immunohistochemistry results showed that RPS4Y1 is expressed in the villi,decidua,placenta of pregnant women,and placenta of PE patients in the early pregnancy,and is mainly localized in vascular endothelial cells and trophoblasts;2.In the normal foot The relative expression of RPS4Y1 m RNA was detected in both placenta and placenta of preeclampsia.Compared with the normal control group,the relative expression of RPS4Y1 m RNA(0.89±0.05)was significantly increased in PE group(1.54±0.04)(P<0.05).3.WB protein quantification results showed that compared with RPS4Y1 protein in placenta tissue of patients with normal blood pressure in late pregnancy,the amount of RPS4Y1 protein in PE placenta tissue increased significantly,and the results showed statistical significance(P<0.05).4.Interference with RPS4Y1 can promote HUVEC cell migration and angioplasty,without significant effects on cell proliferation and apoptosis.5.H/R treatment can lead to a decrease in the biological function of HUVEC cells and increase the expression of RPS4Y1 protein in HUVEC cells.At the same time,H/R treatment inhibited the activation of the JAK2/STAT3 cell pathway in HUVEC cells.6.The JAK2/STAT3 signaling pathway may be located downstream of the RPS4Y1 gene.Conclusion: 1.RPS4Y1 is mainly located in endothelial cells and trophoblast cells in the early pregnancy.2.The expression of RPS4Y1 in placenta of preeclampsia was significantly higher than that of normal placental tissues.3.Increased expression of RPS4Y1 can inhibit the angioplasty and cell migration of HUVEC cells.The expression of RPS4Y1 gene was affected by oxidative stress.Through the JAK2/STAT3 signaling pathway,the biological function of HUVEC cells was regulated and involved in the development of PE.