脂多糖直接诱导对肺微血管内皮细胞IL-8表达的影响及通过NF-κΒ调控机理的研究  

An research on the expression of IL-8 in PMVEC directly-induced by LPS and its mechanism through the Nuclear Factor-Kappa B

作　　者：顾大勇[1] 王华[1] 刘金标[2] 陈莉[2] 杨季云[2] 马布仁[2] 曾祥元[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所,重庆400000 [2]成都军区总医院临床实验研究所,四川成都610083

出　　处：《四川医学》2004年第9期984-987,共4页Sichuan Medical Journal

摘　　要：目的　探讨脂多糖 (LPS)的直接诱导作用对肺微血管内皮细胞 (PMVEC)IL 8表达的影响及通过核因子κB(NF κB)的调控机理。方法　以 10 0ng/mlLPS刺激PMVEC 0 ,0 .5 ,1,2 ,4,6,8h或 1ng/ml、10ng/ml、10 0ng/mlLPS刺激 1h或6h为检测时相点 ,ELISA、原位杂交试验分别检测的培养液上清中分泌的IL 8及PMVEC内IL 8mRNA的表达 ;凝胶电泳迁移率分析 (EMSA)检验NF κΒ的活化 ;并观察NF κΒ活化抑制对IL 8表达的影响。结果　LPS能显著促进PMVEC表达IL 8,包括促进IL 8mRNA的表达及IL 8的分泌。在时间上mRNA的表达先于IL 8分泌。且LPS的直接诱导能迅速活化NF κΒ ,1h达到高峰 ,后逐渐下降。PDTC能显著抑制NF κΒ的活化及IL 8的表达 (P <0 .0 1)。结论　表明细菌致病因子LPS的直接诱导确能通过促进NF κΒ的活化 ,从而启动IL 8的高效表达和分泌 ,为多形核中性粒细胞 (PMN)的迁移提供必需的物质条件 ,导致肺损伤。Objective To study IL-8 expression of pulmonary microvascular endothelial cells (PMVEC)directly induced by LPS and its mechanism through the Nuclear Factor-Kappa B.Methods When PMVEC is directly excited by 100ng/ml LPS at 0h,0.5h,1h,2h,4h,6h,8h or 1ng/ml,10mg/ml,100ng/ml LPS at 1h or 6h,the expressions of IL-8mRNA in PMVEC and its protein secretion in the supernatant of culture medium were examined by the methods of ISH,ELISA respectively and the activation of NF-κ Β was detected by EMSA.And those were then to be compared with which was inhibited by PDTC.Results LPS could directly act on the PMVEC to promote the IL-8 expression,including promoting the expression of IL-8mRNA and secretion of IL-8 protein,and the mRNA expressions increased before the protein secreted in terms of time.The LPS excitement could also significantly promote the activation of NF-κ Β, which peaked at 1h and then went down.PDTC could significantly decrease the expression of IL-8 and the activation of NF-κ Β.Conclusion The directly-induction of LPS could exactly promote the activation of NF-κ Β,then cause to the expression and secretion of IL-8.SO as to provide the required physical condition for the PMN migration.It could be an important role in inflammatory response induced by LPS.

关 键 词：LPS 核因子ΚB 肺微血管内皮细胞 IL-8 PMN 趋化作用 

分 类 号：R563.8[医药卫生—呼吸系统]