保护素D1对中性粒细胞和人脐静脉内皮细胞黏附性的影响  

The effects of Protectin D1 on the adhesion of neutrophils and human umbilical vein endothelial cells

作　　者：王燕[1] 姚尚龙[1] 舒化青[1] 郭丽娜[1] 伍晓莹[1] 王雅鑫[1] 

机构地区：[1]华中科技大学附属协和医院麻醉科,武汉市430022

出　　处：《临床麻醉学杂志》2013年第1期65-68,共4页Journal of Clinical Anesthesiology

基　　金：湖北省卫生厅项目(JX4C25)

摘　　要：目的探讨保护素D1(Protectin D1,PD1)对LPS刺激人脐静脉内皮细胞(HUVECs)与中性粒细胞(PMNs)之间黏附性的影响及其机制。方法黏附性检测实验分为五组:P2,P20,P200、LPS和对照组(C组)。除C组外,各组HUVECs均用1μg/mlLPS作用6h后,P2、P20、P200分别加入经2、20、200ng PD1预处理的BCECF-AM标记的PMNs,C组和LPS组加入未经PD1处理的BCECF-AM标记的PMNs,作用30min后,用荧光显微镜对黏附在HUVECs上BCECF-AM标记的PMNs进行检测;用流式细胞仪检测PMNs表面淋巴细胞相关抗原-1(LFA-1)表达;Western blot检测HUVECs上细胞间黏附分子-1(ICAM-1)的表达。结果 P2、P20、P200和LPS组的荧光强度值明显高于C组(P<0.05),P2、P20和P200组的PMNs与HUVECs之间黏附率明显低于LPS组(P<0.05),并呈浓度依赖性,浓度越高黏附率越低。P2、P20、P200和LPS组表面抗人LFA-1表达量明显高于C组(P<0.05),P20和P200组明显低于LPS组(P<0.05)。不同浓度的PD1呈浓度依赖性地降低LFA-1的表达浓度,越高表达越低,而对ICAM-1的表达有降低趋势但差异无统计学意义。结论 PD1可降低PMNs与HUVECs之间黏附率,可能是与降低PMNs表面LFA-1表达有关。Objective To investigate the effects and mechanism of PD1 on the adhesion of neutrophils （PMNs） and human umbilical vein endothelial cells （HUVECs） induced by lipopolysaceharide （LPS）. Methods Human umbilical vein endothelial cells （HUVECs）cultured in vitro was randomly divided into five groups for the experiment of PMNs-HUVECs adhesion. There were group P2, group P20, group P200 ,group LPS, group control（group C）. LPS （1 tzg/ml） was used to stimulate HUVECs for 6 h except the group C. BCECF-AM labeled PMNs were added into the HUVECs μmonolayer for another 30 min. PMNs adhesion was detected by fluorescence microscope. Flow cytometry was applied to assess lymphocyte function-associated antigen （LFA-1） expression. Western blotting was used to detect the intercellular adhesion molecuh-1 （ICAM-1） expression. Results Compared with group C, the fluorescence intensity in group P2, P20, P200 and LPS were significantly increased （P〈0. 05）. Compared with group LPS, the PMNs-HUVECs adhesion of P2, P20. P200 were significantly decreased （P〈 0.05）, and the PD1 inhibited the adhesion of PMNs to HUVECs in a concentration-dependent manner. Compared with group C, the expression of LFA-1 in group P2 ,P20 ,P200 and LPS were obviously increased （P〈0. 05）, while the group P20 and P200 were lower than group LPS （P 〈0. 05）. PD1 suppressed LFA-1 expression in PMNs （P 〈 0.05） in a concentration-dependent manner without any significant effect on ICAM-1 expression in HUVECs. Conclusion PD1 can inhibit the adhesion of PMNs and HUVECs in a concentration-dependent manner. It may be related to the decrease of LFA-1 expression in PMNs.

关 键 词：保护素D1 中性粒细胞 人脐静脉内皮细胞 细胞黏附性 

分 类 号：R965[医药卫生—药理学]