LPS直接诱导的肺微血管内皮细胞与PMN的粘附作用及核因子κB调控机理的实验研究

AN EXPERIMENTAL STUDY OF ADHESION OF PMVEC AND PMN AND MOLECULAR MECHANISM OF NF-KAPPA B ACTIVATION INDUCED BY LPS

目的 :研究细菌脂多糖 (LPS)诱导的肺微血管内皮细胞 (PMVEC)与多形核中性粒细胞 (PMN)的粘附作用及调控机制 ;方法 :1 0 0ng/mlLPS刺激PMVEC 0h、2h、4h、6h、8h或 1 0ng/ml、50ng/ml、1 0 0ng/mlLPS刺激 6h ,检测PMVEC -PMN粘附率、PMVEC细胞间粘附分子 -1 (ICAM -1 )的表达 ;凝胶电泳迁移率变化分析 (EMSA)方法检测核因子κB(NF -κB)的活化 ,并通过加入Anti ICAM 1抗体或活化阻断剂观察对PMVEC -PMN粘附率的影响 ;结果 :PMVECICAM -1的表达及与PMN的粘附与LPS的刺激呈时相 -剂量依赖方式 ,LPS的刺激迅速活化NF -κB ,60min达到高峰 ,后逐渐下降 ,Anti-ICAM -1抗体、PDTC能显著降低PMVEC -PMN粘附 (P <0 .0 0 1 ) ;结论 :LPS刺激诱导NF -κB的活化 ,启动ICAM -1的合成表达 ,从而导致PMVEC -PMN的粘附增加。

To study the adhesion of pulmonary microvascular endothelial cells(PMVEC) to polymorphonuclear neutrophils(PMN)induced by LPS and its mechanism.Method:To measure the adhesion rate of PMEVC-PMN and ICAM-lexpression stimulated by 100ng/ml LPS at Oh,2h,4h,6h,8h or 10ng/ml,50ng/ml,100ng/ml LPS at 6h and the activation of NF-κB by EMSA.Above results were compared with which was modulated by anti-ICAM-1 or PDTC.Results:The expression of ICAM-1 and the adhesion rate of PMVEC-PMN were highly depended on the stimulation about the dose and time of LPS.And NF-κB was rapidly activated by LPS stimulation and peaked at 1h,then lowed.The anti-ICAM-1 or PDTC could significantly decreased the adhesion of PMVEC-PMN( P<0.01 ).Conclusion:The activated NF-κB by LPS starts up ICAM-1 expression and increases the adhesion of PMVEC-PMN.