论文部分内容阅读
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.
OBJECTIVE: To investigate the release of inflammatory mediators and their mechanisms during bronchial epithelial cells and eosinophil co-culture.Methods: Flow cytometry (FCM) was used to quantitatively compare eosinophils, bronchial epithelial cells and their combination The release of monocyte chemoattractant protein (MCP) -1 in culture supernatants and the influence of p38MAPK inhibitor SB203580 intervention were investigated. The effect of eosinophils on the bronchus MCP-1 gene expression in epithelial cells and the inhibitory effect of SB203580 on MCP-1 expression.Results: After eosinophil activation, MCP-1 gene expression was significantly up-regulated in bronchial epithelial cells; eosinophils and bronchial epithelial cells MCP-1 protein release in the co-culture supernatant was significantly increased (1266 ± 127 ng / L vs 134 ± 25 ng / L, P <0.001). Paraformaldehyde-fixed eosinophils did not release MCP- 1, but it still increased the release of MCP-1 in bronchial epithelial cells [(773 ± 48) ng / L vs (107 ± 15) ng / L, P <0.001]; p38MAPK inhibitor SB203580 Can effectively inhibit eosinophil activated bronchitis MCP-1 gene expression in epithelial cells significantly reduced the release of MCP-1 during the co-culture of normal eosinophils and paraformaldehyde-fixed eosinophils with bronchial epithelial cells [(1335 ± 115) ng / L vs 481 ± 42 ) and / or (868 ± 89) ng / Lvs (239 ± 26) ng / L respectively, P <0.001] .Conclusion: Eosinophils can induce bronchial epithelial cells to express MCP-1 through p38 MAPK signaling pathway, Sexual airway inflammation.