核因子kappaB的诱骗性寡核苷酸对肿瘤坏死因子-α诱导的细胞间粘附分子-1表达的影响

Effect of decoy oligonucleotides of Nuclear Factor-kB on intercellular adhesion molecule-1 expression in human vascular endothelial cells

目的 探讨人脐静脉内皮细胞中 ,核因子kappaB (NF κB)的诱骗性寡核苷酸对肿瘤坏死因子(TNF) α诱导的细胞间粘附分子 (ICAM) 1表达的影响。方法 采用培养的人脐静脉内皮细胞株ECV30 4 ,脂质体介导法转染寡核苷酸 ,以流式细胞仪检测转染效率 ,电泳迁移率转换试验检测诱骗性寡核苷酸与NF κB转录因子的亲和力及特异性 ,并以RT PCR和流式细胞仪测定其对TNF α诱导的粘附分子ICAM 1表达的影响。结果 诱骗性寡核苷酸能与NF κB特异结合 ,并可显著下调TNF α诱导的人脐静脉内皮细胞ICAM 1的表达。结论NF κB的诱骗性寡核苷酸可显著下调NF κB调控的与动脉粥样硬化相关的粘附分子的表达 ,为核酸药物防治动脉粥样硬化的临床应用提供一些前期资料。

Objective To investigate the effects of decoy oligonucleotides (ODNs) of NF-κB on expression of adhesion molecules induced by TNF-α in vitro. Methods The affinity and specificity of decoy ODNs of NF-κB to the transcription factor NF-κB were detected by electrophoretic mobility shift assay (EMSA). Intracellular uptake of FITC-labeled oligonucleotides was detected by fluorescence active cell sort (FACS). Adhesion molecules expression was determined by RT-PCR at RNA level, FACS at protein level. Results The data showed that decoy ODNs of NF-κB can bind with the transcription factor NF-κB and has strong affinity and specificity. The decoy ODNs can downregulate the expression of intercellular adhesion molecule-1 (ICAM-1) stimulated by TNF-α in human vascular endothelial cells (HUVECs). Conclusion These observations demonstrate that the decoy oligonucleotides of NF-κB can inhibit NF-κB-mediated activities and downregulate the expression of some gene product as ICAM-1 involved in atherosclerosis.