Gene therapy for anastomotic stenosis after arterial reconstruction

自从神经元超囊病发生后，神经元超囊病的形成就被认为是关键的，目前的研究的目的是建立基因转移到神经元的方法，并确定重要因素（s）调节神经元形成中的细胞循环。为了第一个目的，我们应用了腺病毒载体将LacZ基因转化为大鼠Carotima的新钙细胞，是由气球损害诱导的，并发现了超过1x 10 y D19 y D1 PFU/mL的病毒载体允许高效率转移到LacZ基因到新神经细胞。Miyata等人通过上面的方法管理了突变平台衍生出的生长因子受体的基因，并展示了对大鼠carotid neoinimal细胞的一种显著抑制。要研究细胞周期在新的良性形成中的作用，在气球-免疫大鼠胡萝卜素细胞蛋白（Rb）中的表达被西方块分析评估，自从Rb释放的磷酸化从E2 F转录因子中绑定到Rb，细胞周期中的关键事件是什么。在气球损伤后的一天里，Rb的波段转移，Rb的磷酸化指标，被观察到。Rb的磷酸化，在受伤2天后成为最大值，当在动脉媒体中检测到最大值细胞复制时。动脉壁上的cyclin D表达在气球损伤后也被观察到，在损伤后14天的研究生表达增加了，并表明Rb磷酸化和cyclin D表达之间的干扰。Meanwhile，第27页，细胞循环抑制剂，在未损坏的动脉壁中表现出来。第27页的表达在受伤后的7天内再次受到直接调节，但在受伤后的7天内再次增加，这些结果建议第27页被抑制的循环素D在第7天之后调节，并对受伤的动脉细胞的Rb进行调节。

Since neointimal hyperplasia is known to be critical in formation of anastomotic stenosis after arterial bypass operation, the purposes of the present study were to establish the method of gene transfer into neointima and to determine the important factor(s) regulating cell cycle in neointimal formation. For the first purpose, we applied adenovirus vectors containing LacZ gene to neointima of rat carotid artery, which was induced by balloon injury, and found that more than 1x10ィイD19ィエD1 PFU/mL of virus vector promoted high efficiency transfer of LacZ gene to neointimal cells. Miyata et al administrated the gene of mutant platelet-derived growth factor receptor to the rat carotid neointimal cells by the above method, and showed a significant suppression of the neointimal proliferation. To study the role of cell cycle in neointimal formation, expression of retinoblastoma protein (Rb) in balloon-injured rat carotid artery was assessed by western blot analysis, since phosphorylation of Rb releases the E2F transcription factor from its binding to Rb, which is a critical event in cell cycle. At 1 day after balloon injury, band shift of Rb, which indicates phosphorylation of Rb, was observed. The phosphorylation of Rb, became to be maximal at 2 days after injury, when maximal cell replication was detected in arterial media. The expression of cyclin D in arterial wall was also observed after balloon injury, though the expression increased gradually by 14 days after injury, indicating a discrepancy between Rb phosphorylation and cyclin D expression. Meanwhile, p27, cell cycle inhibitor, was expressed in uninjured arterial wall. The expression of p27 was down-regulated once immediately after injury but increased again from 7 days after injury These results suggested that p27 suppressed cyclin D after- day 7 and regulated phosphorylation of Rb of injured arterial cells.