Role of Calpain in Diabetic Endothelial Dysfunction

钙蛋白酶在糖尿病内皮功能障碍中的作用

基本信息

批准号：
7413884
负责人：
Rosario G Scalia
金额：
$ 3.74万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-30 至 2008-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7413884
关键词：
Accounting Acute Adherence Adhesiveness Anti-Inflammatory Agents Anti-inflammatory Attenuated Biochemical Biology Blood Vessels Calcium Calpain Cardiovascular Diseases Cardiovascular system Cataract Cell Adhesion Molecules Cell physiology Cell surface Chronic Clinical Complement Factor B Condition Cysteine Protease Data Development Diabetes Mellitus Diabetic Angiopathies Disease Electrophoretic Mobility Shift Assay Endopeptidases Endothelial Cells Endothelium Event Exhibits Family Functional disorder Gelshift Analysis Gene Mutation Genes Glucose Goals Human Hyperglycemia Immunoblot Analysis Immunohistochemistry Immunoprecipitation In Vitro Incidence Inflammatory Inflammatory Response Intercellular adhesion molecule 1 Kinetics Knockout Mice Knowledge Leukocyte Rolling Leukocytes Life Link Literature Mammals Measurement Measures Methodology Microcirculation Modeling Molecular Morbidity - disease rate Mus Nitric Oxide Non-Insulin-Dependent Diabetes Mellitus Nuclear Patients Pattern Peptide Hydrolases Phosphorylation Physiological Physiology Play Post-Translational Protein Processing Post-Translational Regulation Process Property Protein Isoforms Protein Kinase C Proteins Rate Rattus Regulation Reporting Role Signal Pathway Techniques Technology Testing Up-Regulation Vascular Cell Adhesion Molecule-1 Vascular Diseases Vascular Permeabilities Western Blotting calpain 10 diabetic experience human NOS3 protein in vivo indexing intravital microscopy m-calpain mortality novel novel therapeutics prevent response transcription factor vascular inflammation

项目摘要

Cardiovascular disease accounts for an overwhelming proportion of the morbidity and mortality suffered by patients with all forms of diabetes. Hyperglycemia is considered an important etiologic factor that serves as the initial trigger for diabetic vascular complications. Clinical evidence demonstrates that cardiovascular disease in diabetic patients increases as a function of the duration of hyperglycemia and that exposure of the vasculature to elevated ambient glucose causes generalized endothelial dysfunction, accelerates the atherosclerotic process, and impairs important functions of the microcirculation. As a consequence of hyperglycemia, the diabetic vasculature experiences abnormal inflammatory processes characterized by impaired release of nitric oxide (NO) and increased endothelial adhesiveness. Calpains are a family of calcium-dependent proteases, which have been recently implicated in acute inflammatory disorders of the cardiovascular system. In preliminary studies, we have made the novel observation that inhibition of calpain activity preserves release of endothelial NO and attenuates inflammatory leukocyte-endothelium interactions in the microcirculation during hyperglycemia. Our data strongly support a role for calpains in the pathophysiology of diabetic vascular disease, suggesting a potentially beneficial effect of calpain inhibition in diabetes. Therefore, we propose to study the role of calpains in the inflammatory response of the microcirculation during acute and chronic hyperglycemia. We will test the hypothesis that calpains cause vascular inflammation during hyperglycemia by studying whether: (1) calpain activity is abnormally increased; (2) inhibition of calpain activity prevents inflammatory events in the microcirculation; (3) increased calpain activity downregulates the eNOS enzyme leading to loss of physiologic levels of NO; (4) activation of calpains increases endothelial cell surface expression of pro-inflammatory adhesion molecules via upregulation of NF-IcB activity. We will utilize the following in vivo and in vitro cell physiology techniques: intravital microscopy, culture of microvascular endothelial cells under static and flow conditions, NO measurements in vivo and in vitro, immunohistochemistry, Western blot analysis, and gel shift assays. By these means, the studies in this proposal will elucidate important and novel mechanisms underlying microvascular dysfunction in diabetes. This information should provide a framework for developing new therapeutic strategies for the treatment of the life-threatening disorder, diabetes mellitus.

心血管疾病在发病率和死亡率中占绝大多数患有各种形式的糖尿病的患者。高血糖被认为是一个重要的病因，糖尿病血管并发症的最初触发因素。临床证据表明，心血管糖尿病患者的疾病随着高血糖持续时间和暴露于高血糖的情况而增加。血管系统升高的环境葡萄糖会导致广泛的内皮功能障碍，加速动脉粥样硬化过程，并损害微循环的重要功能。由于高血糖时，糖尿病血管系统会经历异常炎症过程，其特征是一氧化氮（NO）释放受损和内皮粘附性增加。 Calpains 是一个家族钙依赖性蛋白酶，最近与急性炎症性疾病有关心血管系统。在初步研究中，我们做出了新的观察，即抑制钙蛋白酶活性可保留内皮 NO 的释放并减弱炎症性白细胞与内皮细胞的相互作用高血糖期间的微循环。我们的数据强烈支持钙蛋白酶在糖尿病血管疾病的病理生理学，表明钙蛋白酶抑制对糖尿病血管疾病的潜在有益作用糖尿病。因此，我们建议研究钙蛋白酶在炎症反应中的作用。急性和慢性高血糖期间的微循环。我们将检验钙蛋白酶引起的假设通过研究是否：（1）钙蛋白酶活性异常来评估高血糖期间的血管炎症增加； (2)抑制钙蛋白酶活性可预防微循环中的炎症事件； (3) 钙蛋白酶活性增加会下调 eNOS 酶，导致 NO 生理水平下降； (4) 钙蛋白酶的激活增加内皮细胞表面促炎性粘附的表达通过上调 NF-IcB 活性来调节分子。我们将利用以下体内和体外细胞生理学技术：活体显微镜、静态和流动下微血管内皮细胞培养条件、体内和体外 NO 测量、免疫组织化学、蛋白质印迹分析和凝胶转变分析。通过这些方式，本提案中的研究将阐明重要且新颖的机制糖尿病潜在的微血管功能障碍。该信息应提供一个框架开发新的治疗策略来治疗危及生命的疾病——糖尿病。