APS复发性流产患者血小板的活化及其对内皮细胞的影响

Platelet activation and its effect on endothelial cells in patients with recurrent spontaneous abortion with antiphospholipid syndrome

目的:探讨APS复发性流产患者血小板活化状态及其对内皮细胞活化的作用。方法:流式细胞术分别检测23例APS患者和31例正常早孕人群血小板CD62P和CD40L的表达水平。洗涤血小板悬液与人脐静脉内皮细胞(HUVECs)共培养6 h,ELISA法分析HUVECs培养上清IL-8的水平,荧光定量PCR法检测HUVECs细胞ICAM-1的mRNA水平。结果:APS组血小板CD62P和CD40L阳性表达百分率为(10.27±3.76)%和(13.26±5.21)%,较正常早孕组显著升高(P<0.05);APS组血小板悬液能显著促进HUVEC细胞ICAM-1基因表达水平和分泌IL-8水平。特异性CD40L单抗可以显著抑制HUVEC细胞表达ICAM-1水平和分泌IL-8水平。结论:APS流产患者外周血血小板活化程度增加,血小板-内皮细胞反应参与APS导致高凝、炎症和流产的病理过程。

Objective:To investigate the effects of platelets on adhesion molecules expression and chemokines secretion of cultured endothelial cell in patients with recurrent spontaneous abortion with antiphospholipid syndrome.Methods:Peripheral blood platelets CD62P and CD40L expression from 23 APS patients and 31 normal early pregnancy people were tested with flow cytometry respectively.Washed platelets suspension were co-cultured with cultured human umbilical cells for 6 hours.The expression level of Intercellular adhesion molecule1(ICAM-1)in HUVECs and soluble interleukin-8(IL-8)produced by HUVECs were measured with FQ-PCR method and ELISA technique respectively.Results:The expression levels of CD62P molecule were(10.27±3.76)%and levels of CD40L molecule were(13.26±5.21)%on peripheral blood platelets surface in APS patients,which increased significantly compared with those in normal early pregnancy people(P<0.05).The production level of IL-8 and the mRNA level of ICAM-1 of HUVECs stimulated by the activated platelets were higher in APS group and their effects could be inhibited by addition of CD40L-specific monoclonal antibody.Conclusion:Platelets in peripheral blood of APS patients were at higher activated state and endothelial response induced by platelets may be one of the cause of hypercoagulability,inflammation and abortion in APS patients.