Nitric Oxide-Superoxide in Lipid Induced Vascular Diseas

描述(申请人提供)：内皮依赖性异常 血管内一氧化氮生物利用度降低所致的血管松弛 WALL是与高脂血症相关的血管疾病的关键组成部分， 高血压、糖尿病、肥胖症和动脉粥样硬化 在动脉粥样硬化的发病机制中起重要作用。虽然有很多 血管壁中NO生物利用度降低的潜在原因， 减少了NO的生成，或通过与 超氧阴离子可能起到关键作用。基因治疗接近于 动脉粥样硬化可能包括系统性地将基因输送到肝脏进行治疗 风险因素或局部给药至血管壁以提高非生物利用度 增加血液流量，促进新血管形成或限制细胞增殖 在血管壁上。一种局部应用一氧化氮合酶的基因治疗方法 由于一氧化氮具有多效性，基因到血管壁具有优势 血管系统中的抗动脉粥样硬化作用。此外，一个更好的 超氧化物歧化在多种血管内皮细胞损伤中的作用 动脉粥样硬化的分期可能允许超氧化物的治疗作用 歧化酶过表达有待研究。在本提案中，我们将测试 根据假设a)单个一氧化氮合酶亚型可能具有不同的 正常人和疾病患者血管反应性改变的特点 B)腺相关病毒载体可用于转移 一氧化氮合酶基因转移到血管壁导致血管的长期改变 无炎症反应性和c)一氧化氮合酶长期过表达。 高胆固醇血症兔血管壁改善内皮功能 依赖血管松弛和延缓动脉粥样硬化的进展 通过超氧化物歧化酶基因转移增强对超氧化物歧化酶的清除作用可能增加一氧化氮 生物利用度和逆转脂质诱导的内皮功能障碍。这些 实验将确定哪种一氧化氮合酶亚型在改变血管方面最好 功能，检测eNOS长时间表达对血管内皮细胞功能的影响 用AAV载体构建兔颈动脉，检测一氧化氮合酶的作用 过度表达对动脉粥样硬化进展的影响及阐明其作用 超氧化物歧化在不同阶段血管内皮细胞功能障碍中的作用 动脉硬化。

DESCRIPTION (provided by applicant): Abnormal endothelium dependent vasorelaxation due to reduced nitric oxide bioavailability in the blood vessel wall is a key component of vascular disease associated with hyperlipidemia, hypertension, diabetes mellitus, obesity and atherosclerosis and may be important in the pathogenesis of atherosclerosis. While there are many potential causes of decreased NO bioavailability in the blood vessel wall, decreased NO generation or increased NO degradation via interaction with superoxide anions may play a pivotal role. Gene therapy approaches to atherosclerosis may include systemic delivery of genes to the liver to treat risk factors or local delivery to the vessel wall to enhance NO bioavailability to augment blood flow, enhance new vessel formation or limit cell proliferation in the vessel wall. A gene therapy approach utilizing local delivery of NOS gene to the vessel wall has advantages as nitric oxide has pleiotropic anti-atherogenic effects in the vasculature. In addition, a better understanding of the role of superoxide in endothelial dysftinction in various stages of atherosclerosis may allow the therapeutic effects of superoxide dismutase overexpression to be explored. In this proposal we will test the following hypotheses a) individual NOS isoform may have distinct characteristics for altering vascular reactivity in the normal and diseased blood vessel wall, b) adeno-associated virus vectors can be used to transfer the NOS gene to the vascular wall resulting in long term alteration of vascular reactivity without inflammation and c) long term overexpression of NOS in the blood vessel wall of the hypercholesterolemic rabbit will improve endothelium dependent vasorelaxation and delay the progression of atherosclerosis and d) increased scavenging of superoxide via SOD gene transfer may increase NO bioavailability and reverse lipid-induced endothelial dysfunction. These experiments will determine which NOS isoform is best at altering vascular function, examine the functional effect of prolonged expression of eNOS in the rabbit carotid artery using AAV vectors, examine the effect of NOS overexpression on progression of atherosclerosis and elucidate the role of superoxide in the pathogenesis of endothelial dysfunction in various stages of atherosclerosis.