摘要
目的研究核转录因子κB(NFκB)在高糖、肿瘤坏死因子(TNFα)、白细胞介素1β(IL1β)诱导ECV304血管内皮细胞损害中的作用。方法以构建含NFκB抑制物IκBα突变体(IκBαM)的重组腺病毒感染ECV304细胞,用Western印迹、凝胶电泳迁移率实验(EMSA)、四甲基偶氮唑盐(MTT)等方法研究NFκB在高糖、TNFα、IL1β介导血管内皮细胞损害中的作用。结果TNFα可诱导ECV304细胞的IκBα降解和NFκB的激活(P<0.01),而对ECV304/IκBαM细胞则无此作用;高糖可导致ECV304细胞NFκB的激活(P<0.05),而对ECV304/IκBαM细胞则无此作用;单独使用高糖、TNFα、IL1β导致ECV304细胞活力的降低(均P<0.01),而ECV304/IκBαM细胞可抵抗这些因素对细胞的损害(均P<0.05)。结论高糖、TNFα、IL1β可致血管内皮细胞活力降低,IκBα能有效抑制上述有害因素导致的ECV304细胞NFκB的过度活化,抵抗内皮细胞的损害。抑制NFκB活性可能有助于保护血管内皮细胞的功能。
Objective To investigate the role of NF-κB in high glucose （HG） and cytokines （TNF-α and IL-1β）-induced impairment of ECV-304 cells （ vascular endothelial cell line）. Methods Recombinant adenovirus containing NF-κB supper-repressor IκBαM with mutant IκBα was constructed. Western blot, electrophoretic mobility shift assay （ EMSA ） and thiazolyl blue viability assay were applied in this study. Results TNF-α-induced IκBα degradation and NF-κB activation （ P〈0.01 ） were found in ECV-304 cells but not in ECV-304/IκBαM cells ; and HG-induced NF-κB activation （ P〈0.05 ） was also found in ECV-304 cells but not in ECV-304/IκBαM cells intected with IκBαM recombinant adenovirus. The viability of ECV-304 cells incubated with HG, TNF-α or IL-1β was obviously decreased （ all P〈0.01 ） , but ECV-304/IκBαM cells could resist the effect of HG and cytokines on cell viability （ all P〈0.05 ）. Conclusion HG and cytokines （ TNFα, IL-β） can decrease the viability of vascular endothelial cells. IκBα can effectually inhibit the excessive activation of NF-κB in ECV-304. Inhibition of NF-κB activity can protect vascular endothelial cells from the cytotoxicity of HG and the cytokines.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2005年第4期310-312,共3页
Chinese Journal of Endocrinology and Metabolism
基金
国家"十五"863资助项目(2001AA217121)
国家自然科学基金资助项目(30070297)
