IκB-ζ高表达在调控脂多糖诱导细胞炎症反应中的作用

Effect of the high expression of IκB-ζ in lipopolysaccharide-induced inflammation in cells

目的探讨IκB-ζ在调控脂多糖(lipopolysaccharide,LPS)诱导细胞炎症反应中的作用。方法利用免疫共沉淀技术筛选人脐静脉内皮细胞(human umbilical vein enlothelial cells,HUVECs)中IκB-ζ的结合蛋白,采用蛋白质质谱分析方法鉴定IκB-ζ的互作蛋白;应用合成的NfκBiz质粒转染HUVECs并进行LPS干预,Western检测NfκB2(P100,P52)表达量,同时应用q PCR检测炎症因子,如白细胞介素-1(interleukin-1,IL-1)、IL-6及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达。结果蛋白质质谱分析结果显示NfκB2与IκB-ζ结合互作;HUVECs细胞中IκB-ζ过表达并经LPS干预后NfκB2(P100,P52)蛋白表达明显下降。q PCR测试结果显示与LPS组相比IκB-ζ高表达组IL-1及TNF-α表达明显下调(P<0.05),而IL-6表达明显上调(P<0.05)。结论在LPS干预的HUVECs细胞中过表达IκB-ζ能够与NfκB2(P100,P52)蛋白亚基结合,并对细胞的炎症反应进行调节,降低IL-1及TNF-α的表达同时增加IL-6表达。

Objective To investigate the effect of IκB-ζ in lipopolysaccharide（LPS）-induced inflammation in cells. Methods Proteins interacting with IκB-ζ were screened by co-immunoprecipitation and identified by protein mass spectrometry analysis method. NfκBiz-predominated plasmid was used to transfect human umbilical vein endothelial cells（HUVECs） induced by lipopolysaccharide. The expression of NfκB2（P100,P52） was detected by Western blotting. The levels of mRNA of Interleukin-1,Interleukin-6 and TNF-α were detected by q PCR. Results Protein mass spectrometry analysis results showed that IκB-ζ interacted with NfκB2. The overexpression of NfκB2（P100,P52） was substantially decreased in HUVECs induced by lipopolysaccharide. The levels of mRNA of IL-1 and TNF-α were decreased（P〈0. 05）; however,IL-6 was increased（P〈0. 05）. Conclusion The overexpression of IκB-ζin HUVECs induced by lipopolysaccharide could regulate inflammation by combining with the subunits of NfκB2（P100,P52）,and meanwhile decrease the expression of mRNA of IL-1 and TNF-α,and increase the expression of IL-6.