HDAC1 Regulation of Endothelial NO

项目总结 过量的钠摄入量每年导致数十亿美元的医疗费用。2010年，165万人死亡 心血管疾病的风险归因于高盐饮食。此外，钠的摄入量与 与肾功能和死亡率有关。高盐饮食会增加心血管和肾脏的损伤 氧化应激和损伤内皮细胞一氧化氮(NO)信号。高盐饮食的机制 导致NOS3解偶联和增加NADPH氧化酶2(NOX2)的表达和活性仍然难以捉摸。 NOS3解偶联的特征是一氧化氮合酶依赖的活性氧物种(ROS)增加和 减少一氧化氮合酶依赖的一氧化氮。拟议研究的中心假设是高盐分会增加 内皮细胞HDAC1通过翻译后修饰(去乙酰化)解偶联NOS3，并增加 NOX2通过转录机制表达和活性。为了研究HDAC1在NOS3中的作用 分离正常和高盐饲养的大鼠肾内动脉和肾内皮细胞。 将评估HDAC1活性以及NOS3解偶联和NOS3乙酰化对HDAC1的依赖。我们 将血管内皮细胞HDAC1基因敲除(VEHDAC1KO)小鼠用于研究HDAC1的必要性 在高盐中，没有干扰任何信号。延髓旁肾单位的准备将用于评估无信号。 在肾内的小动脉。为了确定哪些NOS3赖氨酸对HDAC1去乙酰化敏感，我们 将利用包含特定赖氨酸(K)到精氨酸(R)或谷氨酰胺(Q)突变的NOS3结构。CdH5- CreERT2-Nox2KO小鼠将被用来评估内皮NOX2在高盐干扰NO信号转导中的作用 在肾内的小动脉。我们将使用VEHDAC1KO小鼠来阐明HDAC1在高盐增加中的作用 NOX2复合表达。芯片定量聚合酶链式反应将用于评估HDAC1对组蛋白修饰的影响 NOX2复合体基因的启动子。该项目的长期目标是了解 HDAC1在血管内皮细胞NO信号转导中的作用，以开发心血管疾病的新疗法。

PROJECT SUMMARY Excess sodium intake accounts for billions of healthcare dollars every year. In 2010, 1.65 million deaths from cardiovascular disease were attributed to a high salt diet. Furthermore, sodium intake directly correlates with kidney function and mortality. A high salt diet causes cardiovascular and renal damage by increasing oxidative stress and impairing endothelial nitric oxide (NO) signaling. The mechanisms by which a high salt diet leads to NOS3 uncoupling and increased NADPH oxidase 2 (Nox2) expression and activity remains elusive. NOS3 uncoupling is characterized by an increase in NOS-dependent reactive oxygen species (ROS) and a decrease in NOS-dependent NO. The central hypothesis to the proposed studies is that high salt increases endothelial HDAC1 which uncouples NOS3 via post-translational modification (deacetylation) and increases Nox2 expression and activity via a transcriptional mechanism. In order to study the role of HDAC1 in NOS3 uncoupling, intra-renal arteries and renal endothelial cells will be isolated from normal and high salt fed rats. HDAC1 activity as well as HDAC1 dependence of NOS3 uncoupling and NOS3 acetylation will be assessed. We will generate vascular endothelial HDAC1 knockout (VEHDAC1KO) mice to investigate the necessity of HDAC1 in high salt disrupted NO signaling. The juxtamedullary nephron preparation will be used to assess NO signalling in small intra-renal arteries. In order to identify which NOS3 lysines are susceptible to HDAC1 deacetylation, we will utilize NOS3 constructs containing specific lysine (K) to arginine (R) or glutamine (Q) mutations. Cdh5- CreERT2-Nox2KO mice will be used to assess the role of endothelial Nox2 in high salt disrupted NO signaling in small intra-renal arteries. We will use VEHDAC1KO mice to elucidate the role of HDAC1 in high salt increased Nox2 complex expression. ChIP-qPCR will be utilized to assess the effect of HDAC1 on histone modifications at the promoters of the genes of the Nox2 complex. The long-term goal of this project is to understand the role of HDAC1 in endothelial NO signaling in order to develop novel therapies for cardiovascular disease.