GRO-α regulation in airway smooth muscle by IL-1β and TNF-α:: role of NF-κB and MAP kinases
GRO-α regulation in airway smooth muscle by IL-1β and TNF-α:: role of NF-κB and MAP kinases
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DOI：
10.1152/ajplung.00384.2005
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发表时间：
2006-07-01
影响因子：
4.9
通讯作者：
Chung, Kian Fan
中科院分区：
文献类型：
--
作者：
Issa, Razao;Xie, Shaoping;Chung, Kian Fan
气道平滑肌细胞(ASMC)是一种炎性趋化因子的来源，可促进呼吸道炎症反应。我们研究了细胞因子诱导ASMC产生CXC趋化因子生长相关癌基因蛋白-α(Gro-α)以及MAPK和NF-kappa B通路的作用。ASMC从人的呼吸道培养，生长至融合，生长停止后暴露于细胞因子IL-1β和TNF-α。在IL-1β(0.1 ng/ml)或TNF-α(1 ng/ml)作用下，Gro-α的释放以剂量和时间依赖的方式增加50倍或5倍。辅助性T细胞2型细胞因子IL-4、IL-10和IL-13不影响Gro-α的释放。IL-1α和TNF-α也可诱导Gro-α基因的表达。IL-1β刺激的ASMC上清液对中性粒细胞具有趋化作用，这种作用可被抗Gro-α封闭抗体抑制。IKK-2的抑制剂AS-602868和ERK的抑制剂PD-98059抑制Gro-α的释放和表达，而JNK的抑制剂SP-600125则减少Gro-α的释放，但不影响Gro-α的表达。P38MAPK抑制剂SB-203580对此无作用。AS-602868对IL-1β和肿瘤坏死因子-α诱导的p65DNA结合有抑制作用，而PD-98059和SP-600125无此作用。染色质免疫沉淀实验显示，IL-1β和肿瘤坏死因子-α可促进p65与Gro-α启动子的结合，而AS-602868可抑制这一作用。IL-1β和TNF-α刺激的ASMC Gro-α的表达受包括核因子-kappaB激活、ERK和JNK通路在内的独立信号通路的调节。ASMC释放的Gro-α参与中性粒细胞的趋化。
Airway smooth muscle cells ( ASMC) are a source of inflammatory chemokines that may propagate airway inflammatory responses. We investigated the production of the CXC chemokine growth-related oncogene protein-alpha ( GRO-alpha) from ASMC induced by cytokines and the role of MAPK and NF-kappa B pathways. ASMC were cultured from human airways, grown to confluence, and exposed to cytokines IL-1 beta and TNF-alpha after growth arrest. GRO-alpha release, measured by ELISA, was increased by > 50-fold after IL-1 beta ( 0.1 ng/ml) or 5-fold after TNF-alpha ( 1 ng/ml) in a dose- and time-dependent manner. GRO-alpha release was not affected by the T helper type 2 cytokines IL-4, IL-10, and IL-13. IL-1 alpha and TNF-alpha also induced GRO-alpha mRNA expression. Supernatants from IL-1 beta-stimulated ASMC were chemotactic for neutrophils; this effect was inhibited by anti-GRO-alpha blocking antibody. AS-602868, an inhibitor of IKK-2, and PD-98059, an inhibitor of ERK, inhibited GRO-alpha release and mRNA expression, whereas SP-600125, an inhibitor of JNK, reduced GRO-alpha release without effect on mRNA expression. SB-203580, an inhibitor of p38 MAPK, had no effect. AS-602868 but not PD-98059 or SP-600125 inhibited p65 DNA-binding induced by IL-1 beta and TNF-alpha. By chromatin immunoprecipitation assay, IL-1 beta and TNF-alpha enhanced p65 binding to the GRO-alpha promoter, which was inhibited by AS-602868. IL-1 beta- and TNF-alpha-stimulated expression of GRO-alpha from ASMC is regulated by independent pathways involving NF-kappa B activation and ERK and JNK pathways. GRO-alpha released from ASMC participates in neutrophil chemotaxis.