Oxidative Stress and Vascular HO in Diabetes

糖尿病中的氧化应激和血管 H2O2

• 批准号: 8011295
• 负责人: Nader G. Abraham
• 金额: $ 24.18万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 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/8011295
• 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; DNA; Development; Diabetes Mellitus; Diabetic Angiopathies; Diabetic mouse; 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; 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; overexpression; prevent; promoter; response; stem; type I and type II diabetes; type I diabetic; vascular endothelial dysfunction

DESCRIPTION (provided by applicant): 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, increases 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-1 prevents endothelial apoptosis by decreasing cellular heme content, increasing CO and bilirubin levels and decreasing EC-SOD and DNA degradation. 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 O2- and improved vascular response. In addition, 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-1 and 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) act 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 overexpress 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.

描述（由申请人提供）：宏观和微血管疾病是1型和2型糖尿病患者发病率和死亡率的主要原因。自由基的释放增加证明了内皮功能障碍，增加了粘附分子和凋亡。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，这可以通过药理学介导的HO-1上调来预防。我们的初步结果表明，HO-1的上调通过降低细胞血红素含量，增加CO和胆红素水平并降低EC-SOD和DNA降解，从而阻止内皮细胞凋亡。该提案的目的是阐明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/或药理方法来实现长期血管保护的使用。我们的假设是，HO-1基因转移将通过增加血红素降解并随后增加CO和胆红素合成来提供强大的血管保护。 CO和胆红素的水平升高将导致EC-SOD和ENOS增加，O2和改善的血管反应降低。此外，线粒体HO-1水平的升高将增加线粒体传输载体，减少线粒体ROS并防止细胞色素C的释放以及Caspase 9的激活。我们计划使用遗传探针（逆转录病毒LXSN LINEDIVIRAL，以及腺病毒）以及腺病毒和腺病毒）和遗传自发的糖尿病小鼠（点头）小鼠。我们将使用功能丧失的HO-2（ - / - ）和功能获得，HO-1和HO-2基因转移到HO-2（ - / - ）来破译每个基因在血管保护。以下特定目标将检验该假设。 （1）确定使用逆转录病毒载体有选择地增加HO-1的遗传干预是否在糖尿病大鼠和NOD小鼠中提供血管保护，以及这种作用是否归因于CO，胆红素或两者兼而有之。我们将检查HO-1得出的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，而CO，胆红素或两者都是通过EC-SOD的增加而对血管保护的强制性。 ，eNOS，O2和Inos的减少。 （3）确定HO-1基因表达（CO和胆红素）作用于促凋亡和抗凋亡蛋白的外在和内在途径的机制。这将检查HO-1在线粒体和细胞质中产生的CO和胆红素的作用。 （4）评估使用细胞特异性启动子（VE-CAD）靶向内皮细胞过表达HO-1基因的慢病毒载体是否足以抵消糖尿病诱导的血管损伤。该提议的方法是新颖的。它将首次允许对HO-1在血管保护中的功能以及用于治疗I型糖尿病的创新基因靶向疗法的深入分析。