Oxidative Stress and Vascular HO in Diabetes

糖尿病中的氧化应激和血管 H2O2

• 批准号: 7630645
• 负责人: Nader G. Abraham
• 金额: $ 5.6万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7630645
• 关键词: Adenovirus Vector; Animals; Apoptosis; Apoptotic; Attenuated; Bilirubin; Biliverdine; Blood Vessels; Carbon Monoxide; Cardiovascular Diseases; Cardiovascular system; Cell Adhesion Molecules; Cell Death; Cells; Complication; Cytoplasm; Cytoprotection; Depth; Development; Diabetes Mellitus; Diabetic Angiopathies; Diabetic mouse; Elevated Bilirubin; Endothelial Cells; Experimental Diabetes Mellitus; Free Radicals; Functional disorder; Gene Expression; Gene Targeting; Gene Transfer; Genes; Genetic; Germ Lines; Goals; Heme; Human; Hyperglycemia; In Vitro; Inbred NOD Mice; Inflammation; Injury; Insulin; Insulin-Dependent Diabetes Mellitus; Isoenzymes; Laboratories; Lead; Lentivirus Vector; Mediating; Mitochondria; Morbidity - disease rate; Mus; Oxidative Stress; Oxygenases; Pathway interactions; Patients; Pharmaceutical Preparations; Physiological; Protein Overexpression; Proteins; Publishing; Rattus; Relative (related person); Research Proposals; Resistance; Retroviral Vector; Role; System; Testing; Time; Transgenic Mice; Up-Regulation; Vascular Diseases; Vascular Endothelium; Work; base; case control; caspase-9; cobaltiprotoporphyrin; cytochrome c; diabetic; diabetic rat; gain of function; gene therapy; heme oxygenase-1; heme oxygenase-2; improved; in vivo; innovation; loss of function; mortality; mouse model; novel; prevent; promoter; response; stem; type I and type II diabetes; type I diabetic; vascular endothelial dysfunction

Macro and microvascular diseases are the principal causes of morbidity and mortality in patients with type 1 and type 2 diabetes. Endothelial dysfunction, as evidenced by the increased release of free radicals, increase adhesion molecules and apoptosis. Previous work by us and others has shown that a decrease in HO-1 gene expression enhances apoptosis and vascular injury which can be prevented by pharmacological mediated upregulation of HO-1. Our preliminary results indicate that upregulation of HO-1prevents endothelial apoptosis by decreasing cellular heme content, increasing CO and bilirubin levels and decreasing EC-SOD and DMAdegradation. The goal of this proposal is to elucidate the mechanism by which HO-1 prevents diabetes-mediated endothelial dysfunction and to explore the use of genetic and/or pharmacological approaches to achieve long-term vascular protection. Our hypothesis is that HO-1 gene transfer will provide powerful vascular protection by increasing heme degradation and subsequently increasing CO and bilirubin synthesis.The increased levels of CO and bilirubin will result in an increase in EC-SOD, and eNOS, a decrease in Oa"and improved vascular response. In addition, and increase in mitochondrial HO-1 levels will increase mitochondrial transport carrier, decrease mitochondrial ROS and prevent release of cytochrome c and activation of caspase 9. We plan to test this hypothesis using genetic probes (retroviral LXSN lentiviral, and adenoviral) and in genetically spontaneously diabetic mice (NOD) mice. We will use the loss-of-function HO-2 (-/-)and gain-of-function, HO-1and HO-2 gene transfer to HO-2 (-/-) to decipher the role of each gene in vascular protection. The following specific aims will test the hypothesis. (1)To determine whether genetic intervention, using retroviral vectors, to selectively increases HO-1 provides vascular protection in diabetic rats and NOD mice, and whether this effect is due to CO, bilirubin or both. We will examine the effect HO-1 derived CO and bilirubin on EC-SOD, eNOS and O2" levels. (2) HO-2 (-/-) and NOD mice will be used to test the hypothesis that HO-1 gene transfer can provide vascular protection and that CO, bilirubin or both are obligatory for the vascular protection via increase in EC-SOD, eNOS, decrease in O2~ and iNOS. (3)To determine the mechanism whereby HO-1 gene expression (CO and bilirubin) on the extrinsic and intrinsic pathway of pro-apoptosis and anti-apoptotic proteins. This will examine the role of CO and bilirubin produced by HO-1 in both the mitochondria and cytoplasm. (4) To evaluate whether lentiviral vectors targeting endothelial cells using cell specific promoter (VE-CAD) to pverexpress HO-1 gene is sufficient to offset diabetes-induced vascular injury. This proposal is novel in its approach. It will allow for the first time, an in-depth analysis of the function of HO-1 in vascular protection and for the development of innovative gene-targeting therapies for the treatment of type I diabetes.

大血管和微血管疾病是糖尿病患者发病和死亡的主要原因。 1型糖尿病和2型糖尿病内皮功能障碍，如通过增加释放游离 自由基，增加粘附分子和凋亡。我们和其他人以前的工作表明， HO-1基因表达的降低可增强细胞凋亡和血管损伤， 通过药理学介导的HO-1上调。我们的初步结果表明， HO-1通过降低细胞血红素含量、增加CO和 胆红素水平和降低EC-SOD和DMA降解。本提案的目的是阐明 HO-1预防糖尿病介导的内皮功能障碍的机制，并探讨HO-1在糖尿病中的作用。 使用遗传和/或药理学方法实现长期血管保护。我们 假设HO-1基因转移将通过增加血红素提供强大血管保护 降解和随后增加的CO和胆红素合成。CO和胆红素水平的增加， 胆红素将导致EC-SOD和eNOS的增加，Oa "的减少和血管改善。 反应此外，线粒体HO-1水平的增加将增加线粒体转运 载体，减少线粒体ROS并防止细胞色素c的释放和半胱天冬酶9的活化。 我们计划使用基因探针（逆转录病毒LXSN、慢病毒和腺病毒）来验证这一假设， 遗传自发性糖尿病小鼠（NOD）。我们将使用丧失功能的HO-2（-/-）， 功能获得性，HO-1和HO-2基因转移到HO-2（-/-），以破译每个基因在 血管保护以下具体目标将检验这一假设。(1)To确定是否 使用逆转录病毒载体进行遗传干预，选择性地增加HO-1， 保护糖尿病大鼠和NOD小鼠，以及这种效果是否是由于CO，胆红素或两者兼而有之。我们将 检测HO-1诱导的CO和胆红素对EC-SOD、eNOS和O2-水平的影响。(2)HO-2（-/-）和 NOD小鼠将用于测试HO-1基因转移可以提供血管保护的假设 CO、胆红素或两者均通过增加EC-SOD、eNOS O2~-、iNOS减少。(3)To确定HO-1基因表达（CO和 胆红素）对促凋亡和抗凋亡蛋白的外在和内在途径的影响。这将 检测HO-1在线粒体和细胞质中产生的CO和胆红素的作用。(4)到 评估使用细胞特异性启动子（VE-CAD）靶向内皮细胞的慢病毒载体是否 过量表达HO-1基因足以抵消糖尿病引起的血管损伤。这一建议是新颖的， 其方法。这将首次对HO-1在血管内皮细胞中的功能进行深入分析。 保护和开发创新的基因靶向疗法治疗I型糖尿病 糖尿病