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（通过应用程序证明）：该提案的总体目标是中心假设：2型IC小鼠中的冠状动脉功能障碍是由树突状细胞（DC）激活而产生的，随后表达了Internferon Gamma（IFN）特定的目标和每个目标都有2个假设。在2型糖尿病（DB/DB）和CD11C-DTR小鼠之间，在此模型中，DC的“有条件为null”。 wt]，但DC的疾病无效或未用白喉毒素水平处理的小鼠炎性细胞因子（干扰素伽玛IFN和TNF）在用Diptheria毒素治疗后DB/DB/DTR中会降低DB/DB/DTR的小鼠。 WT X CD1C-DTR; DC耗尽的小鼠（DB/DB X CD11C-DTR），将T细胞和巨噬细胞的激活减少到DB/DB/DB/DB/DB或WT X CD1C-DTR。 DC的激活发展为T细胞和巨噬细胞的激活，导致2型糖尿病的冠状动脉毒物中炎症细胞因子释放。 CD4TM1MAK中具有靶向突变的C57BL/6背景上的小鼠模型在CD4+ T细胞发育中显着阻塞）内皮依赖性血管扩张反应是氧化应激（超氧化物的产生，蛋白质硝基化），与2型类型2型相比，降低了糖尿病DB/DB小鼠。 CD11b -DTR这是一个模型，而在CD11b的控制下，人二毒素受体[DTR]会导致循环单核细胞的压缩（超氧化物的产生，蛋白质硝化）与2型糖尿病DB/dB小鼠相比，DTR将与同窝室中的瘦肉控制（db/db x cd11b-dtr）相媲美。由体外显微镜，荧光和电子磁共振共振（EPR）分析（O2？挥之不去生产和Western印迹以评估所有菌株和十字架中的关键蛋白质的表达。 T细胞和ED血管炎症，导致2型糖尿病。炎。 公共卫生相关性：我们的支撑性研究将导致血管炎症引发，从而通过解密机制进行一些特定的原理，从而引发感染性炎症性血管和微血管疾病的螺旋，我们的目的是探索中心假设，即树突状细胞（DCS）具有Interferon interferon Gamma的激活， （IFN）在2型糖尿病中驱动导致内皮功能障碍的事件（IFN），导致内皮功能障碍。产生上治疗功能障碍，如果您未经治疗的话，在糖尿病血管和微血管疾病中达到高潮，会有助于维持适当的矿体功能，并防止病理性刺激性疾病的疾病促进疾病的病理性疾病。 。