绿脓菌素通过PKC-NF-κB信号传导通路诱导人NCI-H292细胞表达IL-8

目的探讨绿脓菌素(PCN)对人气道上皮细胞株(NCI-H292细胞)表达IL-8的诱导作用及通过蛋白激酶C(PKC)和核因子κB(NF-κB)的调控机制。方法采用ELISA法对PCN诱导NCI-H292细胞分泌IL-8进行分析,应用Western blot检测NF-κΒ的蛋白表达,观察PKC和NF-κB阻断剂对IL-8表达的影响。结果PCN可促进NCI-H292细胞IL-8的分泌,而且具有明显的量效关系。PKC阻断剂钙感光蛋白(Cal C)及NF-κB阻断剂二硫代氨基甲酸吡咯烷(PDTC)均能抑制IL-8的表达(P<0.01)。PCN能直接诱导并迅速活化NF-κB,60~90 min表达明显。结论PCN可能通过PKC信号通路促进NF-κB的活化,从而启动IL-8的高效表达。

Expression of IL-8 induced by Pyocyanin in NCI-H292 cells by PKC-NF-κB signal pathway

Aim To study IL-8 expression of human airway epithelium-like NCI-H292 cells directly induced by Pyocyanin and its mechanism through protein kinase C(PKC) and nuclear factor-kappa B(NF-κΒ).Methods ELISA methods were performed to test the expression of IL-8 in NCI-H292 cells infected by Pyocyanin.Western blot was employed to examine the expression of NF-κB protein.In addition,NCI-H292 cells were cultured together with PKC inhibitor,calphostin C or NF-κΒ inhibitor,pyrrolidine dithiocarbamate(PDTC),respectively before Pyocyanin infection,then the expression of IL-8 and NF-κΒ was assayed using the above methods.Results Pyocyanin was able to induce IL-8 protein secretion in NCI-H292 cells and had marked concentration-dependent relations.Pyocyanin could also significantly promote the activation of NF-κB,which peaked at 60~90 min.PDTC and calphostin C could significantly decrease the activation of NF-κΒ and the expression of IL-8.Conclutions Pyocyanin can induce IL-8 production.The direct induction of Pyocyanin can promote the activation of NF-κB by PKC signal pathway,then cause the expression and secretion of IL-8.