The Research for Regulation of Vascular Stenosis by Antisense Delivery System

为了阐明支架作为反义载体候选对血管的影响，我们将支架置入兔降主动脉内，并分析了支架置入后血管内平滑肌细胞增殖和表型的空间时间分布。结果表明，支架对动脉壁的局部作用通过血管内皮细胞的一过性和局部增殖、迁移及其表型调控而导致新生内膜的形成。原位杂交证实，在新生内膜诱导的正常主动脉内支架植入过程中，转化生长因子-β的表达也是短暂的。同时，在动脉粥样硬化性主动脉中，支架植入后新生内膜形成较广泛，未见平滑肌细胞去分化。此外，我们还阐明了巨噬细胞和蛋白多糖在调节动脉粥样硬化性血管内支架植入后新生内膜细胞外基质堆积中的重要作用。Nex…此外，我们还利用兔自体静脉移植模型分析了移植静脉疾病的发生机制。免疫组织化学显示移植后新生内膜形成，血管内皮细胞脱分化、增殖。为了验证体外基因导入静脉移植物的可行性，我们利用HVJ-脂质体的方法将报告基因转入静脉移植物中。基因观察至植入后14d。此外，为了调节平滑肌细胞的增殖，我们转移了衰老细胞衍生抑制物1(SDIL)，这是一种被称为抑制细胞增殖的蛋白质。在sdil-taranfered静脉移植物中，新生内膜的形成少于对照组，因此，我们论证了体外基因转染法的可行性。与体内方法相比，体外交易有两个优点：第一，携带转基因基因的载体对体内的影响较小。其次，由于直接针对靶器官，转染法的疗效较好。外科手术中的体外交易有望在未来实现。较少

To elucidate the effect of stent as candidate for antisense carrier on vessels, we implanted intarvascular stent into rabbit descending thoracic aorta and analyzed the spatial chronological distribution of proliferation and phenotypes of smooth muscle cells in stent-implanted aorta. The results demonstrated that the regional effects on arterial wall by stenting leads to neointima formation through transient and regional proliferation and migration of smooth muscle cells and their phenotypic modulations. In situ hybridization clarified the expression of TGF-beta also transient in neointima imduced stent implantation into normal aorta. Meanwhile, in atherosclerotic aorta, neointima formation after stent implantation was more extensive and dedifferentiation of smooth muscle cells were not seen. In addition, we elucidated that macrophages and proteoglycans play important roles in regulating extracellular matrix accumulation in neointima after stent implantation in atherosclerotic aorta.Nex … More t, we analyzed the mechanism of vein graft disease using rabbit autologous vein implantation model. Immunohistochemistry demonstrated neointima formation after the implantation followed dedifferentiation and proliferation of smooth muscle cells. To test the feasibility of ex vivo gene tranfection into the vein graft, we transferred reporter genes into vein graft using HVJ-liposome method. The genes were observed till 14 days after the implantation. Furthermore, to regulate the proliferation of smooth muscle cells, we transferred senescent cell-derived inhibitor 1 (sdil), known as a protein inhibiting cell proliferation. In sdil-taranfered vein grafts, neointima formation was less than control.Thus, we demonstrated the feasibility of ex vivo gene transfection methods. Ex vivo transaction has two advantages over in vivo methods ; first, the influence of the vector carrying transferred gene was less than in vivo. Second, the efficacy of trnasfection is better because of direct trnasfection against targeted organs. Ex vivo transaction in surgical operation were expected in the future. Less