重组腺病毒IκBαM在血管内皮细胞ECV-304中的表达及对NF-κB活性的抑制

Expression of recombinant adenovirus IκBαM in vascular endothelial cells ECV-304 and its inhibitive effect to the activity of NF-κB

目的 检测重组腺病毒IκBαM(AdIκBαM)在血管内皮细胞ECV - 30 4中的表达及肿瘤坏死因子 (TNF -α)诱导下IκBαM的变化与对NF -κB活性的抑制作用。方法 用携有NF -κB抑制物IκBα突变体的AdIκBαM感染ECV - 30 4细胞 ,用West ernblot及EMSA法检测IκBαM的表达及NF -κB活性的变化情况。结果 AdIκBαM在ECV - 30 4细胞中表达且不被TNF -α诱导降解 ,感染IκBαM的ECV - 30 4 /IκBαM细胞在TNF -α刺激下NF -κB无激活 ,而未感染IκBαM的ECV - 30 4细胞经TNF -α刺激后可有NF -κB的过度活化。结论 AdIκBαM能高效扩增并有效感染ECV - 30 4血管内皮细胞 ,且不会被TNF -α诱导而降解 ,能有效抑制血管内皮细胞NF -κB的过度活化 ,抑制NF -κB活性可能保护血管内皮细胞免于TNF

Objective To investigate the expression of recombinant adenovirus IκBαM in vascular endothelial cells ECV-304 and its inhibiting effect to NF-κB Methods Recombinant adenovirus mediated NF-κB supper-repressor IκBαM with mutant IκBα was constructed Western blot and electrophoretic mobility shift assay (EMSA) were applied in this study Results IκBαM could be expressed stably and efficiently in ECV-304 cells and would not degrade by induction of TNF-α EMSA demonstrated that the infected cells showed no activation of NF-κB before and after the treatment of TNF-α,the cells of uninfected and infected with AdIκBα appeared excessive activation of NF-κB Conclusion AdIκBαM can effectually inhibit the excessive activation of NF-κB in ECV-304 cells Inhibition of NF-κB activation can protect vascular endothelial cells from the injury of TNF-α