Reactive Oxygen Species in Vascular Disease

说明(申请人提供)：NAD(P)H氧化酶在心血管疾病中被广泛激活，包括高血压、动脉粥样硬化和糖尿病。这些氧化酶产生的活性氧物种(ROS)与各种形式的高血压的血管松弛、中膜肥厚和新生内膜增生有关。NAD(P)H氧化酶衍生的超氧阴离子(O2-)和其他ROS被认为介导了牵张诱导的信号转导，导致新生内膜增生。我们之前开发了一种基于细胞的gp91-Phox-(NOX2-)氧化酶组装的抑制物，它能够在血管紧张素II的反应中消除血管O2-的产生。目前的提议源于三个主要发现，证明了多组分的氧化酶组装在血管O2-产生中的重要性；(B)我们的基于NOX2的氧化酶组装的细胞表面抑制物抑制O2-的能力，并抑制球囊血管成形术后大鼠颈动脉的新生内膜增殖；以及(C)新的NOX2同系物NOX1和NOX4的上调，以应对球囊损伤。由于NOX1和NOX4似乎是在拉伸后不同程度地参与血管O2-产生的重要的氧化酶同系物，我们将确定对接序列模拟物(它抑制NOX1和NOX4与其他氧化酶亚基的组装)抑制全血管和内皮、平滑肌细胞和成纤维细胞O2的产生和新生内膜增殖的有效性。这些研究将解决这一假设，即NOX1和NOX4在功能上参与了血管拉伸诱导的氧化酶组装和O2-产生，从而导致新生内膜增殖。将测试三个具体目标：(1)开发基于NOX1和NOX4的氧化酶的特异性抑制剂，并在激素诱导的血管NAD(P)H氧化酶激活的体外模型中测试它们；(2)研究单个基于NOx的氧化酶的对接序列在体外血管拉伸诱导的氧化酶活性中的作用；以及(3)确定NOx对接序列在球囊血管成形术诱导的体内新生内膜增生中的作用。相关性：球囊血管成形术后，旨在破坏血管中各种NAD(P)H氧化酶系统的治疗应能显著改善血管通畅性和功能。这些抑制剂还有望在涉及氧化剂的各种疾病过程中提供广泛的用途，包括高血压、糖尿病和动脉粥样硬化。

DESCRIPTION (provided by applicant): NAD(P)H oxidases are broadly activated in cardiovascular diseases, including hypertension, atherosclerosis, and diabetes. Reactive oxygen species (ROS) derived from these oxidases have been implicated in impaired vascular relaxation, medial hypertrophy, and neointimal hyperplasia in various forms of hypertension. NAD(P)H oxidase-derived superoxide anion (O2-) and other ROS are believed to mediate stretch-induced signaling, leading to neointimal hyperplasia. We previously developed a cell-permeant inhibitor of gp91-phox- (nox2-) based oxidase assembly which is capable of abrogating vascular O2- production in response to angiotensin II. The current proposal stems from 3 major findings, demonstrating (a) the importance of the multi-component oxidase assembly in vascular O2- production; (b) the ability of our cell-permeant inhibitor of nox2-based oxidase assembly to inhibit O2- and attenuate neointimal proliferation of the rat carotid artery in response to balloon angioplasty; and (c) the upregulation of novel nox2 homologues, nox1 and nox4, in response to balloon injury. Since nox1 and nox4 appear to be important oxidase homologues involved differentially in vascular O2- production after stretch, we will determine the efficacy of docking sequence mimics (which inhibit nox1and nox4 assembly with other oxidase subunits) to inhibit whole-vessel and endothelial, smooth muscle cell and fibroblast O2- generation and neointimal proliferation. These studies will address the hypothesis that nox1 and nox4 are functionally involved in vascular stretch-induced oxidase assembly and O2- generation, leading to neointimal proliferation. Three specific aims will be tested: (1) to develop specific inhibitors of nox1- and nox4-based oxidases and test them in an in vitro model of hormone-induced vascular NAD(P)H oxidase activation; (2) to investigate the role of docking sequences on individual nox-based oxidases in vascular stretch-induced oxidase activity in vitro; and (3) to determine the role of nox docking sequences in balloon angioplasty-induced neointimal hyperplasia in vivo. Relevance: Therapies aimed at disrupting the various NAD(P)H oxidase systems in blood vessels should substantially improve vascular patency and function following balloon angioplasty. These inhibitors are also expected to provide broad utility in a variety of disease processes involving oxidants, including hypertension, diabetes and atherosclerosis.