摘要
目的:探讨支气管上皮细胞与嗜酸性粒细胞联合培养过程中炎性介质的释放及其机制.方法:应用流式细胞微珠实验(CBA)方法定量比较嗜酸性粒细胞、支气管上皮细胞及其联合培养上清液中单核细胞趋化蛋白(MCP)-1的释放以及p38MAPK抑制剂SB203580干预的影响;用逆转录聚合酶链反应(RT-PCR)分析联合培养过程中嗜酸性粒细胞对支气管上皮细胞MCP-1基因表达的活化及SB203580对MCP-1表达的抑制作用.结果:经嗜酸性粒细胞活化后,支气管上皮细胞中MCP-1基因表达明显上调;嗜酸性粒细胞和支气管上皮细胞联合培养上清液中MCP-1蛋白质释放显著增加[(1266±127)ng/Lvs(134±25)ng/L,P<0.001];多聚甲醛固定后的嗜酸性粒细胞不能释放MCP-1,但其在与支气管上皮细胞联合培养过程中仍可增加支气管上皮细胞释放MCP-1[(773±48)ng/Lvs(107±15)ng/L,P<0.001];p38MAPK抑制剂SB203580可有效抑制嗜酸性粒细胞活化支气管上皮细胞表达MCP-1基因,显著降低正常嗜酸性粒细胞和多聚甲醛固定嗜酸性粒细胞与支气管上皮细胞联合培养过程中MCP-1的释放[(1335±115)ng/Lvs(481±42)ng/L和(868±89)ng/Lvs(239±26)ng/L,P<0.001].结论:嗜酸性粒细胞可通过p38MAPK信号传导通路活化支气管上皮细胞表达MCP-1,调控过敏性气道炎症反应.
AIM： To investigate the release of monocyte chemoattractant protein （MCP）-1 from the co-culture of human bronchial epithelial cells and eosinophils and the related mechanisms.METHODS： MCP-1 in culture supernatant was deter- mined by Cytometric Bead Array （CBA） Kit in flow cytometry to compare MCP-1 release in bronchial epithelial cells and eosinophils cultured alone or together, and investigate the inhibitive effect of SB 203580, a selective inhibitor of p38 MAPK, on MCP-1 release. The reverse chain reaction （RT-PCR） was used to analyze the gene expression of MCP-1 in bronchial epithelial cells during co-cuhure with eosinophils and the effect of SB 203580. RESULTS： The interaction of eosino- phils and bronchial epithelial cells was found to up-regulate the gene expression of MCP-1 in epithelial cells. MCP-I in co-culture supematant of bronchial epithelial cells and eosinophils was elevated significantly [ （ 1266±127） ng/L vs （ 134±25） ng/L, P 〈 0. 001 ]. Fixed eosinophils, which lost the ability to release MCP-1, could also elevate epithelial cells to release MCP-1 [（773±48） ng/Lvs （107±15） ng/L, P〈0.001] in co-culture. SB 203580 could effectively inhibit MCP-1 gene expression of bronchial epithelial cells during co-culture with eosinophils, and decreased the release of MCP-1 in the co-culture of epithelial cells and eosinophils or fixed eosinophils [ （ 1335±115 ） ng/L vs （481±42） ng/L, （868±89） ng/Lvs （239±26） ng/L, P〈 0.001]. CONCLUSION： Eosinophils can activate bronchial epithelial cells to express MCP-1 in co-culture by p38 MAPKpathway so as to regulate the airway inflammatory reaction.
出处
《第四军医大学学报》
北大核心
2006年第8期677-680,共4页
Journal of the Fourth Military Medical University
基金
香港中文大学医学委员会专项研究基金(CUHK4281/04M)
