Effect of transfer of synthetic double-stranded cis-element "decoy" oligodeoxynucleotides(ODNs) on neointimal thickening.

背景-进行性新生内膜增厚和再狭窄仍然是限制经皮冠状动脉腔内成形术或搭桥手术治疗血管闭塞疾病患者的长期疗效的主要关键问题。我们研究了人工合成的双链顺式元件“诱饵”寡核苷酸(ODN)在兔颈动脉球囊损伤后的作用，并观察了这些ODN对血管内膜增厚的影响。方法与结果--用日本血凝病毒(HVJ)-脂质体方法将荧光素异硫氰酸酯(FITC)标记的ODN导入球囊损伤的兔颈动脉，发现荧光核信号在整个中层广泛分布，而直接转移未经修饰的ODN在球囊损伤的兔颈动脉内呈斑片状和散布分布。凝胶迁移率的变化表明，AP-1DNA结合被激活，而AP-1诱骗ODN降低了AP-1DNA结合活性，这是体内与AP-1特异结合的结果。形态计量学分析显示，在血小板衍生生长因子刺激下，AP-1诱骗寡核苷酸可使人动脉内膜面积显著减少，细胞数显著减少。结论AP-1诱骗寡核苷酸体内转染可显著抑制球囊损伤动脉内膜增厚，AP-1有望成为基因治疗减少再狭窄的分子靶点。

Background--Progressive neointimal thickening and restenosis remain major critical problems limiting the long-term efficacy of percutaneous transluminal coronary angioplasty or bypass surgery to treat subjects with vascular occlusive diseases. We examined the role of transfer of synthetic double-stranded cis-element "decoy" oligodeoxynucleotides(ODNs) in case of balloon injured rabbit carotid arteries and effects of these ODNs on neointimal thickening were investigated.Methods and Results--Transfection of flurescein isothiocyante(FITC)-labeled ODNs, using the hemagglutinating virus of Japan(HVJ)-liposome method, resulted in widespread distribution of fluorescent nuclear signals over the entire medial layer, while direct transfer of unmodified FITC-labeled ODNs showed patchy and scattered distribution in balloon injured rabbit carotid arteries. The gel mobility shift revealed that AP-1 DNA binding was activated, and that the AP-1 decoy ODNs reduced AP-1 DNA binding activity, as a result of specific binding affinity to AP-1 in vivo. In morphometric analyses, AP-1 decoy ODNs led to a significant reduction in the neointimal area and a significant reduction in cell number of human aortic smooth muscle cells, under conditions of platelet-derived growth factor stimulation.Conclusions--As AP-1 decoy ODNs transfection in vivo dramatically prevented neointimal thickening in balloon injured arteries, AP-1 may be an useful molecular target for gene therapy to reduce restenosis.