摘要
为探讨在人脐静脉内皮细胞中,核因子κB的反义核酸及诱骗性寡核苷酸联合作用对肿瘤坏死因子α诱导的血管细胞粘附分子1表达的影响,采用培养的人脐静脉内皮细胞株ECV304,分别和联合应用反义核酸和诱骗性寡核苷酸干预,流式细胞仪检测寡核苷酸转染效率,并用逆转录一聚合酶链反应和流式细胞仪测定其对肿瘤坏死因子α诱导的血管细胞粘附分子1的表达。结果发现,联合应用反义核酸及诱骗性寡核苷酸,可使肿瘤坏死因子α刺激的人脐静脉内皮细胞中血管细胞粘附分子1的表达明显下调,且下调幅度显著大于反义核酸或诱骗性寡核苷酸的单独作用。由此提示,核因子κB的反义核酸及诱骗性寡核苷酸可显著下调核因子κB调控的与动脉粥样硬化相关的粘附分子的表达,且联合作用效果强于单独作用,可能为核酸干预防治动脉粥样硬化提供新的思路。
Aim To investigate the simultaneous effects of antisense and decoy oligonucleotides of nuclear factor (NF) κB on expression of adhesion molecules induced by tumor necrosis factor(TNF)-α in vitro. Methods Intracellular uptake of fluorescein isothiocyanate (F1TC) -labeled oligonucleotides was detected by fluorescence active cell sort (FACS) ; Adhesion molecules expression was presented by reverse transcription-polumerase chain reaction (RT-PCR) at RNA level, FACS at protein level. Results Simultaneous incubation with antisense P65 and decoy oligonucleotides (ODN) of NF-κB, the inhibitory effect of VGAM-1 was 59.0%, 41.1% and 35.2% at mRNA level, 85.5%, 44.3% and 49.2% on human vascular endothclial cells (hUVEC) surface compared with control group, decoy group and antisense group. The inhibitory effect of simultaneous incubation with antisense and decoy oligonucleotides was stronger than the effect of incubation with one of them. Conclusion It was demonstrated that a simultaneous administration of antiscnse and decoy oligonucleotides of NF-κB was effective and necessary if a sufficient inhibition of NF-κB mediated activities should be achieved.
出处
《中国动脉硬化杂志》
CAS
CSCD
2004年第3期305-308,共4页
Chinese Journal of Arteriosclerosis
