Role of Cytokine-Induced Inflammation in Endothelial Dysfunction in Diabetes

细胞因子诱导的炎症在糖尿病内皮功能障碍中的作用

• 批准号: 8291824
• 负责人: Michael A HILL
• 金额: $ 37.36万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8291824
• 关键词: Ablation; Affect; Aftercare; Antigens; Blood Vessels; CD4 Positive T Lymphocytes; Cells; Coronary; DNA; Dendritic Cells; Dendritic cell activation; Development; Diabetes Mellitus; Diabetic Angiopathies; Diabetic mouse; Diphtheria Toxin; Disease; Electron Spin Resonance Spectroscopy; Elements; Endothelium; Event; Fluorescence Microscopy; Functional disorder; Funding; Genetic; Goals; Heart Diseases; Human; ITGAM gene; ITGAX gene; Immune; Impairment; In Vitro; Inflammation; Inflammatory; Injury; Interferon Type II; Knockout Mice; Knowledge; Lead; Link; Macrophage Activation; Mediating; Microcirculation; Modeling; Morbidity - disease rate; Mus; Mutation; Myocardial Ischemia; Natural Immunity; Nitric Oxide; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Oxidative Stress; Play; Population; Production; Proteins; RNA; Receptor Activation; Research Personnel; Risk Factors; Role; Smooth Muscle Myocytes; Societies; Stroke; Superoxides; T-Cell Activation; T-Cell Development; T-Lymphocyte; Testing; Therapeutic; Toxin; Tumor Necrosis Factor-alpha; Tunica Adventitia; Vascular Diseases; Vasodilator Agents; Western Blotting; Work; arteriole; cell type; combat; cytokine; db/db mouse; design; diabetes risk; diabetic; diabetic cardiomyopathy; diphtheria toxin receptor; experience; improved; insight; macrophage; monocyte; mortality; mouse model; neutralizing antibody; nitration; novel; novel strategies; prevent; promoter; research study; response; therapeutic development; therapy design; vascular inflammation

DESCRIPTION (provided by applicant): The overarching goal of this proposal is to test the central hypothesis that: coronary dysfunction in type 2 diabetic mice is produced by the activation of dendritic cells (DC) with subsequent expression of interferon gamma (IFN). There are 3 specific aims and each aim has 2 hypotheses. The first aim will causally determine the role of dendritic cell (DC) activation in endothelial dysfunction in coronary arterioles in type 2 diabetes. We propose that 1) Endothelial function will be preserved in coronary arterioles in a cross between type 2 diabetic (db/db) and CD11c-DTR mice, a mouse "conditionally null" for DC. In this model, DCs are depleted after treatment with Diptheria toxin and comparisons will be made to both littermate controls (lean Db/db or wild type [WT], but conditionally null for DC) or mice not treated with Diptheria toxin (and thus containing the resident population of DCs); and 2) Levels of inflammatory cytokines (interferon gamma IFN and TNF) will be reduced in db/db x CD11c-DTR following treatment with Diptheria toxin and comparable to lean littermates, Db/db or WT x CD11c-DTR; and 3) In diabetic mice depleted of DCs (db/db x CD11c-DTR), activation of T cells and macrophages are reduced compared to db/db mice and comparable to lean littermates, Db/db or WT x CD11c-DTR. The second aim will determine if activation of DCs progresses to activation of T cells and macrophages leading to release of the inflammatory cytokines in coronary arterioles in type 2 diabetes. We propose that in a diabetic model with genetic deletion of T cells (db/db x CD4+ -/-; this is a mouse model on a C57BL/6 background with a targeted mutation in Cd4tm1Mak that significantly blocks in CD4+ T-cell development) endothelium-dependent vasodilatory responses are greater, and markers of oxidative stress (superoxide production, protein nitration) are reduced, compared to that in type 2 diabetic db/db mice. Vasodilatory responses of db/db x CD4+ -/- will be comparable to that of littermate, lean control (Db/db x CD4+ -/-) mice. We also propose that in a diabetic model with Diptheria toxin induced deletion of macrophages (db/db x CD11b-DTR; this is a mouse model which expresses the human diphtheria toxin receptor [DTR] under the control of the CD11b promoter. Treatment with diphtheria toxin results in the complete ablation of circulating monocytes), endothelium-dependent vasodilatory responses are greater, and markers of oxidative stress (superoxide production, protein nitration) are reduced, compared to that in type 2 diabetic db/db mice. Responses of db/db x CD11b-DTR will be comparable to that in the littermate, lean control (Db/db x CD11b-DTR) mice. We will use a combination of methodological approaches principally consisting of in vitro microscopy, fluorescence and electron paramagnetic resonance (EPR) analysis of superoxide (O2¿¿) production, real timePCRfor RNA and DNA analyse s, and Western Blotting to evaluate expression of key proteins in all strains and crosses. We believe that this study will provide a new understanding of, and new approaches for, the treatment of vascular injury in type 2 diabetes and related disorders/complications. This work will increase our knowledge of the role of dendritic cells, T cells and macrophage-induced vascular inflammation, which causes vascular dysfunction in type 2 diabetes. These studies will aid development of therapeutic strategies by providing results that hone our understanding of how DC activation affects coronary impairment and improve how we develop optimal therapeutic strategies for harnessing the adaptive impact of latent, innate immunity to combat inflammation. PUBLIC HEALTH RELEVANCE: Our proposed studies will investigate the causes of vascular inflammation initiating the spiral of endothelial dysfunction and culminating in diabetic vascular and microvascular disease by deciphering mechanisms of coronary microvascular dysfunction in type 2 diabetes. Specifically, our goal is to explore the central hypothesis that the activationof dendritic cells (DCs) with subsequent expression of interferon gamma (IFN) drives the events (T cell activation, macrophage activation, vascular inflammation) that result in endothelial dysfunction in type 2 diabetes. The results of this study are expected to contribute to the development of novel adjunctive therapies by identifying mechanisms and key targetable elements within them that activate vascular dendritic cells to produce endothelial dysfunction, which if untreated culminates in diabetic vascular and microvascular disease. New therapies that target vascular inflammation will aid in maintaining proper endothelial function and prevent initiation of the dysfunctional pathological spiral that inexorably evolves towards the pathologica manifestations of diabetes in the macro- and microcirculation.

描述（由应用程序提供）：该提案的总体目标是测试中心假设：2型糖尿病小鼠中的冠状动脉功能障碍是由树突细胞（DC）激活并随后表达干扰素伽马（IFN）产生的。有3个特定目标，每个目标都有2个假设。第一个目标将有可能确定树突状细胞（DC）激活在2型糖尿病中冠状动脉动脉内皮功能障碍中的作用。我们建议1）在2型糖尿病（DB/DB）和CD11C-DTR小鼠之间的十字架中，内皮功能将保存在冠状动脉art中，这是DC的小鼠“有条件无效”。在该模型中，DC在用白喉毒素治疗后会耗尽，并进行比较与同窝窝对照（瘦的db/db或野生型[wt]，但对于DC的有条件为无效）或未用白喉毒素处理的小鼠（因此包含居民的炎症和2级）（如果含有较高的cyma）（Intry and and Intermant）（Intry and and Intry）。 DB/DB X CD11C-DTR用白喉毒素处理后，与瘦同杂的同窝，db/db或wt x cd11c-dtr相当； 3）在DCS（DB/DB X CD11C-DTR）中耗尽的糖尿病小鼠中，与DB/DB小鼠相比，T细胞和巨噬细胞的激活减少，并且与瘦同窝仔，DB/DB或WT X x CD11C-DTR相当。第二个目的将确定DCS的激活是否发展为T细胞和巨噬细胞的激活，导致2型糖尿病中冠状动脉动脉中炎性细胞因子的释放。我们提出，在具有T细胞遗传缺失的糖尿病模型中（DB/DB X CD4+ - / - ；这是CD4TM1MAK中具有靶向突变的小鼠模型，在CD4+ T-Cell发育中显着阻碍了依赖于依赖于氧化的cd4tm1mak的靶向突变，它依赖于氧化剂的血管疾病响应量更大（依赖于氧化）。在2型糖尿病DB/DB小鼠中。 db/db x cd4+ - / - 的血管舒张反应将与窝窝控制，瘦肉控制（db/db x cd4+ - / - ）小鼠相当。 We also propose that in a diabetic model with Diptheria toxin induced deletion of macrophages (db/db x CD11b-DTR; this is a mouse model which expresses the human diphtheria toxin receptor [DTR] under the control of the CD11b promoter. Treatment with diphtheria toxin results in the complete ablation of circulating monocytes, endothelium-dependent vasodilatory responses与2型糖尿病DB/dB小鼠相比，氧化应激的标记（超氧化物产生，蛋白质硝化）是相比的。对超氧化物的产生（O2€）的电子磁共振共鸣（EPR）分析，REALPCRFOR RNA和DNA分析s和Western印迹，以评估所有菌株和十字架中关键蛋白的表达这项工作将增加我们对树突状细胞，T细胞和巨噬细胞诱导的血管感染的作用的了解，这会导致2型糖尿病中的血管功能障碍。这些研究将通过提供结果来帮助制定理论策略的发展，使我们对DC激活如何影响冠状动脉损害的理解，并改善我们如何制定最佳治疗策略来利用潜在的，先天免疫学对抗击炎症的适应性影响。 公共卫生相关性：我们拟议的研究将研究血管炎症的原因，引发了内皮功能障碍的螺旋，并通过冠状动脉疾病和微血管疾病的最终导致冠状动脉微血管功能障碍的解密机制在2型2型糖尿病中引起。具体而言，我们的目标是探索中心假设，即树突状细胞（DC）随后表达干扰素伽马（IFN）的激活驱动事件（T细胞激活，巨噬细胞激活，血管感染），导致2型糖尿病中内皮功能障碍导致内皮功能障碍。预计这项研究的结果将通过鉴定其内部的机制和关键靶向元素来促进新型辅助疗法的发展，从而激活血管树突状细胞以产生内皮功能障碍，如果未治疗的糖尿病血管和微血管疾病中未经治疗的最终导线。靶向血管感染的新疗法将有助于维持适当的内皮功能，并防止功能失调的病理螺旋形启动，该螺旋形螺旋形，可无可避免地朝着宏观和微循环中糖尿病的病理表现发展。