Pregnancy Specific Glycoprotein 9 Change In Serum Of Preeclampsia Patients And Its Regulatory Effect On Vascular Endothelial Cell Function

BackgroundPre-eclampsia（PE）is a hypertensive condition during pregnancy that occurs after 20 weeks of gestation and has an adverse effect on both mother and child.Preeclampsia is estimated to have a global incidence of about 2-8% and is a major cause of maternal and infant morbidity and mortality,with adverse effects on the long-term prognosis of mothers and children.Its pathogenesis is not perfect,and there is no reliable predictive index,so the study of its pathogenesis is very important.Pregnancy-specific glycoprotein（Pregnancy-specific glycoprotein）is a group of proteins secreted by the placental syncytium embryo layer,which is expressed significantly from Pregnancy.It has been reported that the serum PSG levels of patients with spontaneous abortion,ectopic pregnancy,fetal intrauterine growth retardation,fetal intrauterine hypoxia,and fetal intrauterine growth retardation are significantly reduced.PSG9 is a member of its family,but its relationship with preeclampsia is not clear,and the molecular mechanism involved is less reported.Our initial application of mass spectrometry detected late-onset preeclampsia patients serum PSG9 level significantly lower than normal pregnancy patients,and then through the determination of CCK8 cell proliferation,scratches,tube forming experiment and protein immunoblot,fluorescence detection of nitric oxide and calcium imaging technology to research the PSG9 influence on endothelial cell function,and the molecular mechanisms involved,for a deep understanding of the development of preeclampsia provides some new insights.Purpose（1）To elucidate the changes of PSG9 in serum of preeclampsia patients;（2）To explore the effect of PSG9 on endothelium proliferation,migration and tube formation;（3）To investigate the regulatory effect of PSG9 on calcium homeostasis and nitric oxide release of vascular endothelial cells and its mechanism.Methods1.Differential protein in serum of preeclampsia patients was detected by mass spectrometry;2.CCK8 detected the effect of PSG9 on endothelial cell proliferation;3.The effect of PSG9 on endothelial cell migration was detected by scratch test;4.Tubule formation assay was used to detect the effect of PSG9 on the tubule formation ability of endothelial cells;5.The effect of PSG9 on the expression level of related proteins in cells was detected by Western blot;6.The effect of PSG9 on NO production in cells was detected by NO fluorescent probe assay;7.The effect of PSG9 on intracellular calcium ion concentration was detected by calcium ion imaging assay.ResultsThe results of this study showed that the level of PSG9 in serum of preeclampsia patients was significantly lower than that of normal pregnant women.Secondly,PSG9 could promote angiogenesis and migration of human umbilical vein endothelial cells,but had no obvious proliferation,PSG9 enhanced the expression of SOCE channel proteins ORAI1,ORAI2,ORAI3 and e NOS and the production of NO in HUVECs,and the use of calcium store-operated calcium influx inhibitors reversed the effect of PSG9 on the production of eNOS and NO in HUVECs,it is suggested that PSG9 may be involved in up-regulation of calcium channel proteins and the promotion of extracellular calcium influx during calcium channel regulation.ConclusionThese results suggest that PSG9 affects the proliferation,migration and tube formation of Endothelium,up-regulates the expression of SOCE channel Proteins ORAI1,ORAI2 and ORAI3,and may be involved in the regulation of endothelial function via Ca²⁺/ e NOS/ NO signaling pathway.This study provides a theoretical basis for PSG9 as a potential clinical diagnostic marker and therapeutic target for preeclampsia,and provides a new idea for the treatment of this disease.