目的：探讨支气管上皮细胞（BEAS-2B细胞）与中性粒细胞联合培养时细胞间黏附分子1（ICAM-1）、ICAM-3、血管细胞黏附因子1（VCAM-1）等细胞表面黏附分子的产生机制。方法免疫磁珠法提取外周血中性粒细胞，建立中性粒细胞与BE-AS-2B细胞联合培养体系。流式细胞仪（FCM ）检测BEAS-2B细胞和中性粒细胞中ICAM-1、ICAM-3、VCAM-1的表达量，蛋白质印迹检测细胞内NK-κB及p38-MAPK的表达。结果 BEAS-2B细胞和中性粒细胞联合培养时，BEAS-2B细胞表面黏附分子ICAM-1、ICAM-3、VCAM-1的表达量均较单独培养时明显增高（P＜0．05），加入抑制剂（MG-132、SB203580）表达量均明显下降（P＜0．05）；两种抑制剂对BEAS-2B细胞VCAM-1、ICAM-3表达作用比较差异无统计学意义（P＞0．05）；而MG-132对ICAM-1表达的抑制作用比SB203580强（P＜0．05），且在调控ICAM-1表达上两种抑制剂有协同作用。蛋白质印迹显示联合培养组BE-AS-2B细胞内Phospho-IκBα及Phospho-p38-MAPK蛋白水平明显增高（P＜0．05）。结论 BEAS-2B细胞与中性粒细胞联合培养可激活BEAS-2B细胞体内NF-κB及p38-MAPK通路，与ICAM-1、ICAM-3、VCAM-1的表达相关。%Objective To explore the mechanism of adhesion molecules(ICAM-1 ,ICAM-3 ,VCAM-1) upon the interaction be-tween neutrophils and bronchial epithelial cells (BEAS-2B cells) .Methods The system of human bronchial cells co-cultured with human neutrophils was constructed .The FCM method was used to explore levels of ICAM-1 ,ICAM-3 and VCAM-1 on BEAS-2B cells and neutrophils .The Western blot method was used to explore levels of NF-κB and p38-MAPK proteins .Results The levels of ICAM-1 ,ICAM-3 ,VCAM-1 were higher in co-culture system than those in cells cultured singly (P<0 .05) .When treated with inhibitors (MG-132 ,SB203580) ,the levels of ICAM-1 ,ICAM-3 ,VCAM-1 were decreased(P<0 .05) .The ICAM-1 inhibition effect of MG-132 was much better than SB203580(P<0 .05) .The levels of Phospho- IκBα and Phospho-p38-MAPK were higher in co-culture system than those in cells cultured singly (P<0 .05) .Conclusion In co-cultured system ,the signal transduction pathway of NF-кB and p38-MAPK in BEAS-2B can be activated ,which can regulate the release of adhesion molecules .
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