绿脓菌素激活MAPKs、NF-κB信号通路诱导呼吸道上皮细胞IL-8表达  

Involvement of MAPKs and NF-κB pathways in Pseudomonas pyocyanin-induced interleukin-8 expression by human airway epithelial cells

作　　者：冯艳[1] 王芳[1] 李响[2] 王伯瑶[1] 吴琦[1] 

机构地区：[1]四川大学华西基础医学与法医学院感染免疫教研室 [2]四川大学华西医院泌尿外科,成都610041

出　　处：《微生物学报》2005年第1期86-90,共5页Acta Microbiologica Sinica

基　　金：国家自然科学基金 ( 3 0 2 70 688);GMB科研基金 ( 98-681)~~

摘　　要：采用绿脓杆菌培养上清及绿脓菌素刺激人呼吸道上皮细胞株A5 4 9和SPC A 1,用ELISA方法检测细胞IL 8分泌水平 ,并使用免疫印迹 (Westernblot)方法观察绿脓菌素对细胞内重要的炎症信号传导途径NF κB及丝裂原激活蛋白激酶 (MAPKs)的激活作用。实验发现 ,绿脓杆菌培养上清及绿脓菌素可诱导呼吸道上皮细胞株IL 8分泌增加 ,且具有剂量依赖效应。绿脓菌素刺激细胞可使细胞内IκB α发生降解 ,同时使MAPK家族蛋白分子 (ERK1 2、p38、JNK)发生磷酸化。MEK1 2 (ERK1 2激酶 )抑制剂U0 12 6 (10 μmol L)和p38MAPK抑制剂SB2 0 35 80 (10 μmol L)可降低绿脓菌素诱导A5 4 9细胞IL 8的合成。以上结果显示绿脓菌素通过MAPK信号传导通路增强呼吸道上皮细胞IL 8的表达 ;NF κB通路也参与了绿脓菌素调控细胞ILTo investigate the molecular mechanisms of signaling transduction by which Pseudomonas pyocyanin induces IL-8 expression in human airway epithelial cells, A549 and SPC-A-1 cells were challenged with P. aeruginosa conditioned medium or pyocyanin. Chemokine interleukin-8 (IL-8) release from the challenged cells was measured by ELISA, and Western blot was performed to analyze the degradation of IκB-α and the phosphorylation of MAPKs(mitogen-activated protein kinases) in the extracts from cells stimulated with pyocyanin. Both of P. aeruginosa conditioned medium and pyocyanin remarkably increased IL-8 expression by human airway epithelial cells. Degradation of IκB-α was found shortly after A549 cells were stimulated with pyocyanin. Western hybridization analysis also demonstrated that pyocyanin caused phosphorylation of MAPKs including ERK1/2, p38 and JNK in A549 cells. Pretreatment of A549 cells with U0126(10μmol/L), a selective inhibitor of MEK1/2 (ERK1/2 kinase)or with SB203580(10μmol/L), a specific inhibitor of p38 MAPK, diminished the pyocyanin-induced IL-8 production. These findings suggest that Pseudomonas pyocyanin can increase IL-8 expression by human airway epithelial cells through MAPKs signaling pathways and the activation of NF-κB is also involved in this process.

关 键 词：呼吸道上皮细胞 绿脓菌素 丝裂原激活蛋白激酶 核转录因子NF-ΚB 

分 类 号：R378[医药卫生—病原生物学]