粘附分子CD11b与ICAM-1在PMNs跨内皮移行中作用机制与研究.docVIP

粘附分子CD11b与ICAM-1在PMNs跨内皮移行中的作用机制研究 张雪萍 刘新波　任永功　石碧明　李亚丽　 [摘要]目的 探讨粘附分子CD11b与ICAM-1在PMNs跨内皮移行中的作用。方法培养原代人脐静脉内皮细胞于3.0μM孔径大小聚碳酸膜上直至其长成单层，然后移入一种48孔两室趋化板内，LPS分别刺激PMN及HUVEC，取其中一部分分别用CD11b单抗、ICAM-1单抗、CD11b＋ICAM-1单抗处理，PMN移行率通过计数跨越内皮细胞的PMNs数决定。结果 LPS激活PMNs和HUVECs后，PMN跨内皮移行显著增加(168.77%±3.60%,p0.01).，CD11b单抗和ICAM-1单抗可分别减少这种移行至122.90%±4.65% 和114.90%±6.06%。同样，合用两种单抗也近乎产生叠加效应(93.00%±5.21%)。结论 粘附分子CD11b与ICAM-1的表达影响着中性粒细胞的跨内皮移行作用，且这两种分子有协同作用。 关键词 CD11b ICAM-1 多形核白细胞 脐静脉内皮细胞 跨内皮移行 作者单位：暨南大学医学院附属第二医院深圳人民医院 本文受深圳市科技计划项目基金资助　编号 200204004 张雪萍　女　1969年－　博士研究生　副主任医师，研究方向：麻醉与危重病医学 Roles of adhesion molecules　CD11b and ICAM-1 in PMN transmigrating across endothelium. ZHANG Xueping LIU Xinbo REN Yonggong SHI Biming LI Yali　 　 Department of Anesthesiology, ShenZhen people’s Hospital, the second subordinate hospjtal of JiNan Medical college, ShenZhen 518020, China [Abstract] Objective To investigate the roles of adhesion molecules　CD11b and ICAM-1 in PMN transmigrating across endothelium. Method HUVECs were cultured on gelatin treated microporous membranes until an endothelial cell monolayer was grown. Then they were moved into a 48-well transwell chambers with polycarbonate membranes of 3.0μm pore size system . PMNs were isolated from fresh heparinized blood of healthy volunteers by dextran sedimentation followed by Ficoll-HyPaque gradient centrifugation. PMNs and HUVECs were stimulated with LPS(100ng/ml),then treated or not with CD11bMoAb、ICAM-1MoAb、CD11bMoAb+ICAM-1MoAb. The rate of PMNs migration was measured by determining the number of PMNs through endothelial cell monolayers. Result There is an increase in the number of PMN transmigrating across endothelium monolayer following activation of the PMNs and HUVECs by LPS(168.77%?3.60%,p0.01).Pretreatment with anti-ICAM-1 Ab or anti-CD11b Ab resulted in inhibition of transmigration to 122.90%?4.65% and 114.90%?6.06%.When used in combination, two MoAbs inhibited migration in a nearly additive fashion(93.00%?5.21%). Conclusion The expr