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.

宏观和微血管疾病是患者发病率和死亡率的主要原因 类型1和2型糖尿病。内皮功能障碍，自由的释放增加证明了 自由基，增加粘附分子和凋亡。我们和其他人的先前工作表明 HO-1基因表达的降低会增强凋亡和血管损伤，这可以预防 通过药理介导的HO-1上调。我们的初步结果表明上调 通过减少细胞血红素含量，增加CO和 胆红素水平以及降低EC-SOD和DMADEGRADATION。该提议的目的是阐明 HO-1阻止糖尿病介导的内皮功能障碍的机制并探索 使用遗传和/或药理学方法来实现长期血管保护。我们的 假设是HO-1基因转移将通过增加血红素来提供强大的血管保护 降解并随后增加CO和胆红素合成。 胆红素将导致EC-SOD和ENOS增加，OA的减少并改善了血管 回复。另外，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caspase 9的激活。 我们计划使用遗传探针（逆转录病毒LXSN慢病毒和腺病毒）和 遗传自发的糖尿病小鼠（NOD）小鼠。我们将使用功能丧失HO-2（ - / - ）和 功能获得的HO-1和HO-2基因转移到HO-2（ - / - ）以破译每个基因在 血管保护。以下特定目标将检验该假设。 （1）确定是否 使用逆转录病毒载体有选择地增加HO-1的遗传干预提供血管 糖尿病大鼠和点头小鼠的保护，以及这种作用是由于CO，胆红素还是两者兼而有之。我们将 检查HO-1衍生的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 胆红素或两者都是通过增加EC-SOD，ENOS， O2〜和Inos的减少。 （3）确定HO-1基因表达的机制（CO和CO和 胆红素）在凋亡和抗凋亡蛋白的外在和内在途径上。这会 检查HO-1在线粒体和细胞质中产生的CO和胆红素的作用。 （4）至 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 PVERESPRESS HO-1基因足以抵消糖尿病诱导的血管损伤。该提议是新颖的 它的方法。它将首次允许对HO-1在血管中的功能进行深入分析 保护和开发创新的基因靶向疗法，以治疗I型 糖尿病。