The Clinical Study Of Cell-derived Microvesicles And Related Factors In The Pathogenesis Of Preeclampsia

Objective:Preeclampsia(PE),as a complex pregnancy syndrome,its pathogenesis is still unclear,and there is no effective predictor in clinic.In this study,we compared the differences of cell-derived microvesicles(CMVs),CMVs scavenging proteins,platelet(PLT),leukocyte(WBC)and endothelial cell(EC)adhesion levels in peripheral blood of PE women,normal pregnant women and non-pregnant healthy women,as well as their changes before and after delivery.We will try to explore the clinical significance of CMVs and intercellular adhesion in the pathogenesis of PE,in order to find sensitive biomarkers for clinical prediction of PE,and to provide a theoretical basis for the new treatment of PE.Methods:Section one:The number of CMVs,syncytiotrophoblast microvesicles(STBMVs),Annexin V positive(AV+)CMVs,STBMVs,platelet-derived microvesicles(PMVs),endothelial cell-derived microvesicles(EMVs),leukocyte-derived microvesicles(LMVs)and red blood cell-derived microvesicles(RMVs)in peripheral blood of PE women,healthy pregnant women and non-pregnant healthy women were detected by flow cytometry.The changes before and after delivery of PE women and healthy pregnant women were also detected.Section two:The levels of CMVs scavenging proteins Lactadherin,DEL-1 and Annexin V in peripheral blood of PE women,healthy pregnant women and non-pregnant healthy women were detected by ELISA.The changes before and after delivery of PE women and healthy pregnant women were also detected.Section three:The adhesion of PLT to WBC,PLT to EC and the expression of CD62P on platelets in peripheral blood of PE women,healthy pregnant women and non-pregnant healthy women were detected by flow cytometry.Results:Section one(1)The level of CMVs in PE women was 3.3 times higher than that of healthy pregnant women and 4.4 times higher than that of non-pregnant women.After delivery,healthy pregnant women dropped to 15.3%-15.8%of the prenatal level;PE women dropped rapidly to 5.3%-8.7%of the prenatal level.(2)The level of Annexin V positive(AV+)CMVs in PE women was 5.0 times higher than that of healthy pregnant women and 15.9 times higher than that of non-pregnant women.After delivery,healthy pregnant women dropped to 12.4%-15.7%of the prenatal level;PE women dropped to 4.5%-5.3%of the prenatal level.(3)The total STBMVs level in pregnant women increased significantly,PE women were 4.1 times higher than healthy pregnant women and 9.6 times higher than non-pregnant women;healthy pregnant women was 2.4 times higher than non-pregnant women.After delivery,healthy pregnant women dropped to20.3%-23.2%of the prenatal level;PE women dropped to 3.5%-6.0%of the prenatal level.(4)The AV+STBMVs level in pregnant women increased significantly,PE women were 7.7 times higher than healthy pregnant women and 41.1 times higher than non-pregnant women;healthy pregnant women was 5.4 times higher than non-pregnant women.After delivery,healthy pregnant women dropped to17.1%-20.4%of the prenatal level;PE women dropped rapidly to 2.3%-2.9%of the prenatal level.(5)The level of AV+PMVs in PE women was 4.8 times higher than that of healthy pregnant women and 15.5 times higher than that of non-pregnant women.After delivery,healthy pregnant women dropped to 33%of the prenatal level one day after delivery,and returned to the prenatal level three to four days after delivery;PE women dropped to 2.3%-2.9%of the prenatal level.(6)The AV+EMVs level in pregnant women increased,PE women were 2.3 times higher than healthy pregnant women and 6.6 times higher than non-pregnant women;healthy pregnant women was 2.9 times higher than non-pregnant women.After delivery,there was no significant change in healthy pregnant women comparing with the prenatal level;PE women dropped to 38.1%of the prenatal level.(7)The level of AV+LMVs in PE women was 11.6 times higher than that of non-pregnant women.After delivery,there was no significant change in healthy pregnant women and PE women comparing with the prenatal level.(8)The AV+RMVs level in pregnant women increased significantly,PE women were 5.2 times higher than healthy pregnant women and 17.7 times higher than non-pregnant women;healthy pregnant women was 3.4 times higher than non-pregnant women.After delivery,there was no significant change in healthy pregnant women comparing with the prenatal level;PE women dropped to24.3%-29.4%of the prenatal level.(9)The cutoff values of AV+STBMVs,AV+CMVs,total STBMVs and total CMVs for PE predictive diagnosis were 3630/?l,7995/?l,21×10~4/?l and 66.4×10~4/?l during32-33 gestational weeks,respectively.AV+STBMVs have the highest diagnostic value in predicting PE among them.The sensitivity,specificity,positive predictive value and negative predictive value of AV+STBMVs were 85.4%,97.1%,96.7%and82.5%,respectively.Section two(1)Lactadherin increased significantly in PE women,which was 1.6 times higher than that of healthy pregnant women and 2.9 times higher than that of non-pregnant women before delivery.There was no significant change in healthy pregnant women and PE women comparing with the prenatal level within 4 days after delivery.(2)Lactadherin had the highest correlation with AV+CMVs and AV+STBMVs,with correlation coefficients of 0.525 and 0.524,respectively.(3)The cutoff value of Lactadherin for PE predictive diagnosis was 2541 pg/ml during 32-33 gestational weeks.The sensitivity,specificity,positive predictive value and negative predictive value were 57.6%,92.0%,90.5%and 62.2%,respectively.(4)The sensitivity of combined detection of AV+STBMVs and Lactadherin was higher than that of AV+STBMVs alone(97.0%vs 85.4%)and significantly higher than that of Lactadherin alone(97.0%vs 57.6%).The positive predictive value of combined detection of AV+STBMVs and Lactadherin was higher than that of Lactadherin alone the(95.7%vs 62.2%).(5)The level of DEL-1 in peripheral blood of PE women was not significantly different from that of non-pregnant women and healthy pregnant women before delivery.However,after delivery,healthy pregnant women dropped to 74.3%-80.0%of the prenatal level;PE women dropped rapidly to 68.7%-87.5%of the prenatal level.(6)The Annexin V level in peripheral blood of PE pregnant women was not significantly different from that of non-pregnant women and healthy pregnant women before delivery.There was no significant change in healthy pregnant women and PE women comparing with the prenatal level within 4 days after delivery.Section three(1)PLT-WBC adherence rate of PE pregnant women increased,which was 1.6 times higher than that of healthy pregnant women,4.1 times higher than that of non-pregnant women,and healthy pregnant women was 2.5 times higher than that of non-pregnant women.After delivery,the PLT-WBC adherence rate of healthy pregnant women increased to 1.6 times of that before delivery.However,there was no significant change in PE pregnant women within 3 to 4 days after delivery.(2)There was no significant difference in PLT-EC adhesion between pregnant women with PE and non-pregnant women and healthy pregnant women.There was no significant change in PLT-EC adhesion between healthy pregnant women and PE pregnant women after delivery.(3)The expression rate of CD62P on PLT in pregnant women increased significantly,PE women were 3.4 times higher than healthy pregnant women and 7.4 times higher than non-pregnant women;healthy pregnant women was 2.2 times higher than non-pregnant women.Healthy pregnant women dropped to 72.7%of the prenatal level 3-4 days after delivery;PE women dropped to 27.0%-43.2%of the prenatal level.Conclusions:Section one(1)The total CMVs in PE pregnant women during pregnancy was higher than that in normal pregnant women.The increase of AV+CMVs with high expression of phosphatidylserine(PS)on the membrane surface is the most significant,which may be closely related to the pathological changes of PE hypercoagulability.(2)The levels of various AV+CMVs in PE women were higher than those in healthy pregnant women and non-pregnant women,especially in placenta-derived AV+STBMVs.The levels of AV+STBMVs rapidly decreased to normal postpartum level one day after delivery,which was consistent with the rapid remission of PE patients after placenta delivery.It is suggested that the abnormal increase of AV+STBMVs in blood circulation of PE pregnant women may be involved in the pathogenesis of PE.(3)The predictive value of diagnostic PE at 32-33 weeks of gestation were AV+STBMVs,AV+CMVs,total STBMVs,total CMVs in the order of high to low.Section two(1)Lactadherin is strongly correlated with AV+CMVs and AV+STBMVs during pregnancy.It is suggested that CMVs with high PS expression on membrane surface may be the main inducing factor of compensatory increase of Lactadherin.(2)The expression levels of DEL-1 and Annexin V in maternal peripheral blood were relatively constant during pregnancy.Therefore,DEL-1 and Annexin V may have limited clearance effect on CMVs in the pathogenesis of PE.Section threeThe platelet-leukocyte adhesion and platelet activation in the peripheral blood of PE patients may play a certain role in the pathological process of PE microthrombosis and inflammatory reaction.