转录因子NF-kB对粘附分子及血管新生内膜形成的影响

Role of NF-kB in the Vascular Cell Adhesive Molecular and Neointima Formation

目的以调节多种增殖因子的核转录因子NF -kB的反义和诱骗性寡核苷酸为干预药物 ,探讨它们单独或联合作用对大鼠颈动脉球囊损伤后血管增殖反应及粘附分子表达的影响。 方法SD大鼠随机分为七组 ,制作血管球囊损伤模型 ;相应时间点处死动物进行检测。 结果模型组、正义组、Scramble组的血管内膜面积、中膜面积、内膜 /中膜在第 5d增加 ,14d达到高峰 ;反义组、诱骗组、反义组 +诱骗组干预后 ,血管上述病理指标明显改善 (P<0 .0 5 )。血管细胞间粘附分子 (VCAM - 1)mRNA表达在血管损伤后 6h即可检测到 ,3、5、7d后持续表达增加 ,14d后表达降低 ;免疫组化显示VCAM - 1蛋白质表达在 14d达到高峰。反义组、诱骗组、反义组 +诱骗组治疗后 ,与模型组、正义组、scramble组相比 ,VCAM - 1mRNA表达和蛋白合成在各时相点均降低 (P <0 .0 5 )。Westernblot检测核蛋白抽提物 ,显示核因子NF -kBp6 5在血管损伤后 6h有一定蛋白表达 ,1d后蛋白表达明显增加 ,至 7d达高峰 ,14d后蛋白表达略降低。反义组、诱骗组、反义组 +诱骗组干预后 ,各时相点蛋白质表达均减弱 (P <0 .0 5 )。 结论转录因子NF -kB调控VCAM - 1基因表达和蛋白质合成 ;球囊损伤后血管壁细胞增殖在不同时相点有动态变化 ;脂质体介导局部转染反义。

Objective To examine the in vivo effect of NF-kB on ballon-injured vascular cell adhesive molecular-1 (VCAM-1) and vessel proliferation response in the carotid artery of rats. Methods The Sprague-Dawley rats underwent balloon-dilation injury of the left carotid artery. They were divided into 7 groups. Results In model group, sense group and scramble group, the vessel intima area, media area and the intima/media ratio increased after 5d and reached the maximum after 14d. Antisense group, decoy group, antisense plus decoy group improved these observational index ( P <0.05). VCAM-1 mRNA expression were examined 6h after artery injury. The expressions increased continuously after 3,5 and 7d and decreased after 14d. Immunohistochemical studies revealed VCAM-1 protein was maximally positive-stained after 14d. In antisense group, decoy group and antisense plus decoy group, VCAM-1 mRNA expression and protein synthesis decreased in every time point compared with the model group, sense group and scramble group. Western blot studies showed NF-kB p65 was dispersely positive-stained 6h after injury and increased after 1d and reached the peak after 7d, but protein expression was weak after 14d. The antisense group, decoy group and antisense plus decoy group treatment inhibited protein synthesis more significantly than the model group, sense group and scramble group ( P <0.05).Conclusion Nuclear factor NF-kB modulated the gene expression and protein synthesis of VCAM-1. Celluar proliferation in the vessel wall dynamically changes after balloon angioplasty injury. Antisense and decoy oligonucleotide of NF-kB by local lipofectamine transfer inhibit NF-kB activating genes modulatoin, and the combined effect were remarkable than alone.