IKKBeta Mediated Impairment of Endothelial Nitric Oxide Production

IKKβ 介导的内皮一氧化氮生成损伤

基本信息

批准号：
7269389
负责人：
Francis Kim
金额：
$ 26.96万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7269389
关键词：
1-Phosphatidylinositol 3-Kinase Address Adult Agonist Animals Aspirin Atherosclerosis Attenuated Biological Availability Blood Vessels Bone Marrow Transplantation Bradykinin Cardiovascular Diseases Cause of Death Cells Data Development Diabetes Mellitus Diabetic Angiopathies Diet Dominant-Negative Mutation Endothelial Cells Enzymes Fatty acid glycerol esters Functional disorder Genetic Glucose Goals Immune Impairment Individual Inflammatory Insulin Insulin Resistance Laboratories Lipopolysaccharides Liquid substance Mediating Mediator of activation protein Modeling NF-kappa B Natural Immunity Nitric Oxide Nonesterified Fatty Acids Nutrient Obesity Pathogenesis Pathway interactions Patients Phosphoinositide-3-Kinase, Catalytic, Gamma Polypeptide Phosphorylation Physiological Play Production Protein Overexpression Regulation Research Research Personnel Research Proposals Risk Factors Role Serine Signal Transduction Stimulus Testing Therapeutic Toll-Like Receptor Pathway Toll-like receptors Tumor Necrosis Factor-alpha base cytokine disability feeding human TNF protein in vivo in vivo Model insulin signaling mortality mouse model mouse toll-like receptor 4 programs reconstitution research study response shear stress toll-like receptor 4 vascular inflammation

项目摘要

Description (provided by applicant): Cardiovascular disease is the principal cause of death and disability in adults with diabetes. Endothelial dysfunction is a hallmark of diabetic vascular disease and is characterized by decreased bioavailability of nitric oxide (NO). Our long-term goal is to elucidate the mechanism by which insulin resistance reduces NO production in endothelial cells and thereby facilitate the development of therapeutics that can be used to attenuate the impact of diabetes and insulin resistance on cardiovascular disease. The specific hypothesis of this revised research proposal is that IKKbeta a key enzyme in the regulation of the NF-kappaB inflammatory pathway mediates the effect of insulin resistance on NO production. This hypothesis is based on the following experimental observations completed in our laboratory. First, IKKbeta is activated in endothelial cells by free fatty acids (FFA), high glucose, and TNF-alpha all of which have been implicated in the pathogenesis of insulin resistance. Second, activation of IKKbeta is associated with impaired NO production. Third, inhibition of IKKbeta pharmacologically using aspirin or genetically using a dominant negative IKKbeta construct blocks the ability of FFA, TNF-alpha, or glucose to impair NO production. Finally, overexpression of wild-type IKKbeta recapitulates the effect of FFA, glucose, or TNF-alpha to impair NO production. The results of these experiments suggest that IKKbeta activation is both necessary and sufficient to cause impairment of endothelial NO production. These results also indicate that inflammatory pathways play a key role in both nutrient excess and cytokine mediated endothelial insulin resistance. Based on these observations the experimental focus of this proposal is on the role of IKKbeta in mediating impaired NO production. We propose to determine how activation of IKKbeta impairs endothelial NO production in both an endothelial cell culture model and in an in vivo model of insulin resistance. We will next address how mediators of insulin resistance might activate IKKbeta and examine a potential role for innate immunity (Toll Like Receptor) in mediating vascular insulin resistance. The following aims are proposed: Aim 1. To determine the mechanism by which IKKbeta activation impairs NO production in endothelial cells. Aim 2. To determine whether impaired endothelial insulin signaling and NO production are associated with IKKbeta activation in vivo in a mouse model of diet-induced obesity and insulin resistance. Aim 3. To determine whether the Toll Like Receptor pathway is necessary for endothelial activation of IKKbeta during the development of vascular insulin resistance.

描述（由申请人提供）：心血管疾病是糖尿病成人死亡和残疾的主要原因。内皮功能障碍是糖尿病血管疾病的标志，其特征是一氧化氮生物利用度降低（NO）。我们的长期目标是阐明胰岛素抵抗可以降低内皮细胞中没有产生的机制，从而促进可用于减轻糖尿病和胰岛素耐药性对心血管疾病的影响的疗法的发展。这项修订的研究建议的具体假设是，IKKBETA是调节NF-kappab炎症途径的关键酶，介导了胰岛素抵抗对无生产的影响。该假设基于我们实验室完成的以下实验观察结果。首先，Ikkbeta在内皮细胞中被游离脂肪酸（FFA），高葡萄糖和TNF-α都激活，所有这些都与胰岛素抵抗的发病机理有关。其次，Ikkbeta的激活与无生产受损有关。第三，使用阿司匹林或遗传使用主要的负IKKBETA构建体对IKKBETA进行抑制，可以阻止FFA，TNF-Alpha或葡萄糖损害NO生产的能力。最后，野生型ikkbeta的过表达概述了FFA，葡萄糖或TNF-Alpha的影响，以免损害生产。这些实验的结果表明，Ikkbeta激活既需要且足以引起内皮NO产生的损害。这些结果还表明，炎症途径在营养过量和细胞因子介导的内皮胰岛素抵抗中都起着关键作用。基于这些观察结果，该提案的实验重点是Ikkbeta在介导没有生产受损的情况下的作用。我们建议确定IKKBETA的激活如何损害内皮细胞培养模型和体内胰岛素耐药模型中的内皮无产生。接下来，我们将解决胰岛素抵抗的介体如何激活Ikkbeta并研究先天免疫（TOLL类似受体）在介导血管胰岛素抵抗中的潜在作用。提出了以下目的：目标1。确定IKKBETA激活损害内皮细胞中没有产生的机制。目标2。确定内皮胰岛素信号受损和无产生是否与饮食诱导的肥胖症和胰岛素抵抗的小鼠模型中的Ikkbeta激活有关。目标3。确定在血管胰岛素抵抗发展过程中，IKKBETA的内皮激活是否必要。