CAJ | 학술논문

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。
목적：탐토지기관상피세포（BEAS-2B세포）여중성립세포연합배양시세포간점부분자1（ICAM-1）、ICAM-3、혈관세포점부인자1（VCAM-1）등세포표면점부분자적산생궤제。방법면역자주법제취외주혈중성립세포，건립중성립세포여BE-AS-2B세포연합배양체계。류식세포의（FCM ）검측BEAS-2B세포화중성립세포중ICAM-1、ICAM-3、VCAM-1적표체량，단백질인적검측세포내NK-κB급p38-MAPK적표체。결과 BEAS-2B세포화중성립세포연합배양시，BEAS-2B세포표면점부분자ICAM-1、ICAM-3、VCAM-1적표체량균교단독배양시명현증고（P＜0．05），가입억제제（MG-132、SB203580）표체량균명현하강（P＜0．05）；량충억제제대BEAS-2B세포VCAM-1、ICAM-3표체작용비교차이무통계학의의（P＞0．05）；이MG-132대ICAM-1표체적억제작용비SB203580강（P＜0．05），차재조공ICAM-1표체상량충억제제유협동작용。단백질인적현시연합배양조BE-AS-2B세포내Phospho-IκBα급Phospho-p38-MAPK단백수평명현증고（P＜0．05）。결론 BEAS-2B세포여중성립세포연합배양가격활BEAS-2B세포체내NF-κB급p38-MAPK통로，여ICAM-1、ICAM-3、VCAM-1적표체상관。
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 .