NF-κB及p38MAPK信号通路对支气管上皮细胞与中性粒细胞共培养IL-6分泌的调控作用  

Regulation of NF- κB and p38MAPK on IL-6 induced by the co-culture between neutrophils and bronchial epithelial cells

作　　者：唐红卫[1] 王成彬[1] 

机构地区：[1]解放军总医院临床检验科,北京100853

出　　处：《解放军医学院学报》2014年第7期730-732,737,共4页Academic Journal of Chinese PLA Medical School

基　　金：军队医学科研十二五重大专项(CWS12J021);国家科技支撑计划(2013BAI17B05)~~

摘　　要：目的探讨支气管上皮细胞(bronchial epithelial cells,BEAS-2B)与中性粒细胞(neutrophils,NEU)联合培养时IL-6分泌的机制。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BEAS-2B细胞联合培养体系。应用Roche cobas e411检测上清液中IL-6浓度。结果 BEAS-2B和中性粒细胞联合培养时,上清液IL-6浓度为(3 691±482.3)pg/ml,与细胞单独培养[BEAS-2B(313.4±34.7)pg/ml;NEU(219.1±11.3)pg/ml]差异有统计学意义(P<0.001);蛋白质印迹法(Western blotting)结果显示BEAS-2B和中性粒细胞联合培养可激活BEAS-2B细胞内NF-κB及p38MAPK的信号通路;而加入NF-κB通路抑制剂MG-132,可有效抑制上清液中IL-6的分泌[(1 075.3±83.9)pg/ml vs(3 691±482.3)pg/ml,P<0.01];p38MAPK通路抑制剂SB203580亦能抑制IL-6的分泌[(1 532.8±176.1)pg/ml vs(3 691±482.3)pg/ml,P<0.01];且MG-132的抑制效果明显好于SB2035580[(1 075.3±83.9)pg/ml vs(1 532.8±176.1)pg/ml,P<0.01];当联合使用两种抑制剂(MG-132和SB203580)时可进一步减少IL-6的分泌[(353.1±33.5)pg/ml vs(1 075.3±83.9)pg/ml,P<0.01;(353.1±33.5)pg/ml vs(1 532.8±176.1)pg/ml,P<0.01]。结论 BEAS-2B细胞与NEU细胞接触后激活BEAS-2B细胞体内NF-κB及p38MAPK通路,进而调控IL-6的分泌。Objective To investigate the secretory mechanism of interleukin （IL）-6 when bronchial epithelial cells were co-cultured with neutrophils. Methods Neutrophils were isolated by immune-magnetic beads positive selection method and a system of human bronchial epithelial cells co-cultured with human neutrophils was constructed. The level of IL-6 was detected by the Cobas e411. Results The level of IL-6 （3 691 ± 482.3） pg/ml in the supernatant was significantly higher than the cells cultured singly [BEAS-2B （313.4 ± 34.7） pg/ml; NEU （219.1 _＋ 11.3） pg/ml], （P 〈 0.001）. Western blotting suggested that NF- K B and p38-MAPK in BEAS- 2B could be activated when BEAS-2B was co-cultured with neutrophils. The release of IL-6 could be inhibited effectively with MG- 132 （the proteasome inhibitor of NF- K B） [（1 075.3 ± 83.9） pg/ml vs （3 691 ± 482.3） pg/ml,P 〈 0.01]. SB203580, the proteasome inhibitor of p38-MAPK, could also inhibit the release of IL-6 [（1532.8 ± 176.1） pg/ml vs （3691 ± 482.3） pg/ml, P 〈 0.01]. However, the MG-132 performed better than SB203580 in inhibiting the release of IL-6 [（1 075.3 ±83.9） pg/ml vs （1 532.8± 176.1） pg/ml, P 〈 0.01]. The release of IL-6 decreased further when treated with the two inhibitors [（353.1 ±33.5） pg/ml vs （1 075.3 ± 83.9） pg/ml ） P 〈 0.01; （353.1 ± 33.5） pg/ml vs （1 532.8 ± 176.1） pg/ml, P 〈 0.01]. Conclusion The signal transduction pathway of NF - K B and p38MAPK in BEAS-2B can be activated, which can regulate the release of IL-6, when BEAS-2B are co-cultured with neutrophils.

关 键 词：支气管上皮细胞 中性粒细胞 IL一6 信号通路 

分 类 号：R562.25[医药卫生—呼吸系统]