Elafin在肿瘤坏死因子-α诱导的炎性反应中的抗炎机制

The anti-inflammatory mechanism of Elafin in tumor necrosis factor(TNF)-α-induced inflammation reaction

目的探讨Elafin的抗炎机制。方法体外培养人支气管上皮细胞系16HBE细胞,将已构建好的pEG-FP-N1-Elafin载体转染到细胞中,并以空质粒载体作其对照,以外源性肿瘤坏死因子(TNF)-α作为刺激物共同孵育。间接免疫荧光细胞化学法结合激光共聚焦显微镜检测细胞核中核因子(NF)-κB的活性,ELISA检测细胞上清液中炎性介质(选取IL-1β、IL-8作为代表)的含量,免疫共沉淀法结合Western blot检测小泛素样修饰蛋白(SU-MO)-1-p-IκBα含量。结果重组质粒组的细胞中NF-κB的活性和IL-1β、IL-8的生成量与空质粒载体组细胞相比明显降低,而SUMO-1-p-IκBα的含量则显著升高(P<0.01)。结论Elafin能够通过促进IκBα的类泛素化阻止NF-κB的活化,下调前炎性介质的生成,从而发挥其抗炎作用。

Purpose To investigate the mechanism of Elafin anti-inflammatory action.Methods After cultivating 16HBE cells in vitro,the constructed pEGFP-N1-Elafin eukaryotic expression vector was transfected into the cells,with empty plasmid as a vector control.The cells were then incubated with exogenous TNF-α.The NF-κB activity in nuclear was detected by immunofluorescence and laser scanning confocal microscope,the IL-1β and IL-8 production was assayed by ELISA and the SUMO-1-p-IκBα production was assayed by co-immunoprecipitation and Western blot.Results The NF-κB activity,the IL-1β and IL-8 production of the experimental group were significantly lower than those of the empty plasmid group.The SUMO-1-p-IκBα production of the experimental group was significantly higher than that of the empty plasmid group（all P〈0.01）.Conclusion Elafin prevents NF-κB activation via an effect on IκBα sumoylation,and it then reduces proinflammatory cytokine production,thus playing a role of anti-inflammatory.