AMPK, Metabolic and Inflammatory Stress and the Endothelial Cell

AMPK、代谢和炎症应激以及内皮细胞

• 批准号: 8230872
• 负责人: NEIL B RUDERMAN
• 金额: $ 29.99万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8230872
• 关键词: 5' -AMP-activated protein kinase; AMP-activated protein kinase kinase; Affect; Animals; Aorta; Apoptosis; Atherosclerosis; Attenuated; Blood Vessels; Blood capillaries; Caloric Restriction; Capillarity; Cell Adhesion Molecules; Cells; Central obesity; Chemicals; Confidential Information; Cultured Cells; Deacetylase; Disease; Down-Regulation; Dyslipidemias; Endothelial Cells; Endothelium; Enzymes; Event; Exercise; Fatty Acids; Functional disorder; Funding; Glucose; Goals; Grant; Health; Histones; Human; Hyperglycemia; Hypertension; Hypoglycemia; Inflammation; Inflammatory; Insulin Resistance; Ionophores; Ischemia; Language; Liver; Mediating; Metabolic; Metabolic syndrome; Metformin; Mission; Mitochondria; Modification; Mus; Muscle; Non-Insulin-Dependent Diabetes Mellitus; Oxidative Stress; Palmitates; Phenformin; Phenotype; Predisposition; Protein phosphatase; Proteins; Public Health; Rattus; Regulation; Research; Research Design; Research Methodology; Resveratrol; Rodent; Role; STK11 gene; Signal Transduction; Stimulus; Stress; Techniques; Testing; Thrombin; Training; Tumor Necrosis Factor-alpha; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; angiogenesis; atherogenesis; capillary; density; deprivation; human TNF protein; in vivo; inhibitor/antagonist; insight; lipid metabolism; mitochondrial dysfunction; novel; premature; prevent; response

State the application's broad, long-term objectives and specific aims, making reference to the health relatedness of ; the project (i.e., relevance to the mission of the agency). Describe concisely the research design and methods for achieving these goals. Describe v the rationale and techniques you will use to pursue these goals. i In addition, in two or three sentences, describe in plain, lay language the relevance of this research to public health. If the application is funded, this i description, as is, will become public information. Therefore, do not include proprietary/confidential information. DO NOT EXCEED THE SPACE : PROVIDED. The metabolic syndrome has been defined clinically as a disorder characterized by dyslipidemia, hypertension, central obesity and a predisposition to premature atherosclerotic cardiovascular disease and type 2 diabetes. It is also associated with capillary rarefaction and impaired angiogenesis in muscle. In the preceding grant periods, we demonstrated that AMPK is operative in cultured vascular endothelial cells '(HUVEC, HAEC) and that its activation protects them against the apoptosis, inflammation and mitochondrial (dysfunction caused by incubation in media enriched in glucose, palmitate or TNF-alpha. In addition, we demonstrated the existence of a Sirt1/LKB1/AMPK signaling mechanism in various cultured cells and in rat liver in vivo and we have obtained preliminary evidence for its presence in cultured endothelium. In project 1, we will extend this research by carrying out studies with the following objectives: 1) To characterize the roles of Sirtl and LKB1 in mediating AMPK activation by such factors as glucose deprivation, phenformin, and resveratrol in cultured endothelial cells; 2) To determine whether hyperglycemia induces insulin resistance and apoptosis and enhances TNF-alpha - induced adhesion molecule expression in endothelial cells by causing AMPK dysregulation (decreased activity or impaired activation). If as anticipated it does, we would then determine whether this dysregulation involves inhibition pf the Sirt1/LKB1 mechanism [possibly by post-translational oxidative modification of SirT1 (with project 2)], and/or activation of protein phosphatases; and 3) To examine whether AMPK-mediated events in the vasculature are impaired in vivo in mice with endothelial-cell specific deletions of either SirT1 or LKB1. Studies will be done in cre/lox mice that will be generated by Dr. Walsh (Core B), which he will use to study ischemia-induced angiogenesis (Project 3), and Dr. Cohen (Project 2) atherogenesis in the aorta. We will characterize the effects of exercise in these and in control mice together with Dr. Nathan LeBrasseur, a rodent exercise physiologist. In particular, we will determine how SirT1 and LKB1 downregulation affect the increase in muscle capillary density and the decrease in atherogenic changes in the aorta that typically occur in response to exercise in rodents with a metabolic syndrome phenotype. j - ¿ ^These studies will provide the first information about the function of the SirT1/LKB1/AMPK signaling mechanism in the vasculature. They should also provide novel insights as to whether its dysregulation could be a cause of the endothelial cell dysfunction associated with the metabolic syndrome and a target for its therapy. ,

说明该应用程序的广泛、长期目标和具体目标，并参考以下方面的健康相关性 ;项目（即与机构使命的相关性）。简明地描述实现这些目标的研究设计和方法。描述 v 您将用于实现这些目标的基本原理和技术。 i 此外，用两到三个句子，用通俗易懂的语言描述这项研究与公共卫生的相关性。如果申请获得资助，这 我的描述将按原样成为公共信息。因此，请勿包含专有/机密信息。不要超出空间 ： 假如。 代谢综合征在临床上被定义为以血脂异常为特征的疾病， 高血压、中心性肥胖和过早发生动脉粥样硬化性心血管疾病的倾向 2型糖尿病。它还与肌肉中毛细血管稀疏和血管生成受损有关。在 在资助期之前，我们证明了 AMPK 在培养的血管内皮细胞中起作用 '（HUVEC、HAEC），其激活可以保护它们免受细胞凋亡、炎症和线粒体的影响 （在富含葡萄糖、棕榈酸酯或 TNF-α 的培养基中孵育引起的功能障碍。此外，我们 证明了 Sirt1/LKB1/AMPK 信号传导机制在各种培养细胞和大鼠体内的存在 我们已经获得了其存在于培养的内皮细胞中的初步证据。在项目1中， 我们将通过开展具有以下目标的研究来扩展这项研究：1）表征 Sirtl 和 LKB1 在通过葡萄糖剥夺等因素介导 AMPK 激活中的作用， 培养的内皮细胞中的苯乙双胍和白藜芦醇； 2）判断是否有高血糖 诱导胰岛素抵抗和细胞凋亡并增强 TNF-α 诱导的粘附分子 通过引起 AMPK 失调（活性降低或受损）在内皮细胞中表达 激活）。如果确实如此，我们将确定这种失调是否涉及抑制 pf Sirt1/LKB1 机制[可能通过 SirT1 的翻译后氧化修饰（与项目 2 一起）]， 和/或蛋白磷酸酶的激活； 3) 检查 AMPK 介导的事件是否在 内皮细胞特异性缺失 SirT1 或 SirT1 的小鼠体内血管系统受损 LKB1。研究将在由 Walsh 博士（核心 B）培育的 cre/lox 小鼠中进行，他将用它来 研究缺血诱导的血管生成（项目 3）和 Cohen 博士（项目 2）主动脉动脉粥样硬化形成。我们 将与 Nathan LeBrasseur 博士一起描述运动对这些小鼠和对照小鼠的影响。 啮齿动物运动生理学家。特别是，我们将确定 SirT1 和 LKB1 下调如何影响 肌肉毛细血管密度增加和主动脉中通常发生的动脉粥样硬化变化减少 具有代谢综合征表型的啮齿动物对运动的反应。 j - ¿ ^这些研究将提供有关 SirT1/LKB1/AMPK 信号传导功能的第一批信息 脉管系统中的机制。他们还应该提供新的见解来说明其失调是否会导致 是与代谢综合征相关的内皮细胞功能障碍的原因及其靶点 治疗。 ,