MARKERS AND MECHANISMS OF VASCULAR DISEASE IN DIABETES

糖尿病血管疾病的标志物和机制

• 批准号: 7608104
• 负责人: TIMOTHY J LYONS
• 金额: $ 0.04万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7608104
• 关键词: Affect; Antioxidants; Apolipoprotein A-I; Apolipoproteins; Atherosclerosis; Basic Science; Binding; Blood Vessels; C-reactive protein; CETP gene; Cell Adhesion Molecules; Cell membrane; Cells; Characteristics; Cholesterol; Collaborations; Complement; Complex; Complication; Computer Retrieval of Information on Scientific Projects Database; Cross-Sectional Studies; Data; Diabetes Mellitus; Diabetic Angiopathies; Dyslipidemias; End Point; Endothelial Cells; Enzymes; Erythrocytes; Event; Fibrinolysis; Free Radicals; Funding; Goals; Grant; High Density Lipoproteins; Homocysteine; Homocystine; Human; Hydrolase; In Vitro; Incubated; Inflammation; Inflammatory; Injury; Institution; Insulin-Dependent Diabetes Mellitus; Intervention; Kidney Diseases; Lecithin; Lipid Peroxides; Lipoprotein (a); Lipoprotein (a-); Lipoproteins; Low-Density Lipoproteins; Measures; Mediating; Myocardial Infarction; Non-Insulin-Dependent Diabetes Mellitus; Nuclear Magnetic Resonance; Oxidative Stress; Patients; Plasma; Plasminogen Activator Inhibitor 1; Platelet Activating Factor; Process; Production; Randomized; Rate; Research; Research Personnel; Resources; Retinal Diseases; Role; Serum; Serum amyloid A protein; Source; Standards of Weights and Measures; Stroke; TNF gene; Transferase; United States National Institutes of Health; Vascular Diseases; aryldialkylphosphatase; atherogenesis; cell injury; cohort; diabetic; enzyme activity; glycemic control; human CETP protein; in vivo; macrovascular disease; non-diabetic; oxidation; oxidized low density lipoprotein; programs; prospective; protein expression; release of sequestered calcium ion into cytoplasm; type I and type II diabetes; type I diabetic

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our primary goal is to define the roles of dyslipidemia and oxidative stress in accelerating macrovascular disease (atherosclerosis) in diabetes. Microvascular disease (nephropathy and retinopathy) will be studied as secondary end-points. The place of our project in the broad scope of the program is shown in Figure 1. We will continue (and expand) our longitudinal and cross-sectional studies of the DCCT/EDIC cohort of Type 1 diabetic patients. This cohort is the largest and best-characterized group of Type 1 diabetic patients in the world. Though still relatively young, it has provided exciting data during the first funding period of the Program Project (1996-present). To obtain a higher "hard" event rate (myocardial infarction, stroke etc.) and to study the more common form of diabetes, we will add a Type 2 diabetes cohort from the VA Cooperative Trial "Glycemia and Complications in Diabetes, Type 2". This prospective trial commences October 2000, and as in the DCCT, patients will be randomized to intensive and standard glycemic control. The VA trial provides us with the opportunity to study patients during the randomized portion of the study (in the DCCT, randomization finished in 1993). We have obtained interim funding to support our collaboration with the VA trial in its first year. We will also perform basic science studies to investigate hypotheses we have developed over the past four years relating to dyslipidemia, oxidative stress and atherogenesis in diabetes. These studies, in collaboration with the other Projects and Cores and the Coordinating Centers of both studies, will greatly increase our understanding of the complex multi-factorial mechanisms underlying vascular disease in both types of diabetes. Our Specific Aims are: 1. To define the role of dyslipidemia in the vascular complications of Type 1 and Type 2 diabetes. We will perform comprehensive lipoprotein analyses, including Nuclear Magnetic Resonance (NMR) lipoprotein profiles, levels of apolipoproteins, Lipoprotein (a), and studies of lipoprotein-associated enzymes. We hypothesize that dyslipidemia, including both quantitative and qualitative lipoprotein abnormalities which can now be assessed in much greater detail than ever before, mediates endothelial cell injury and activation, contributing to accelerated atherosclerosis in diabetes. We will obtain NMR lipoprotein profiles of selected subjects from the DCCT/EDIC Type 1 diabetes cohort ("Type 1 Cohort") on two occasions, completing a total of seven determinations over the entire DCCT/EDIC study duration (1983-2006). We will also obtain NMR analyses prospectively on two occasions in the VA Cooperative Trial Type 2 diabetes cohort ("Type 2 Cohort") during 2000-2006. We will measure apolipoproteins A1, B, E, and Lp(a). We will determine activities of the following lipoprotein-related enzymes: paraoxonase (PON), Lecithin Cholesterol Acyl Transferase (LCAT), and Platelet Activating Factor Acetyl Hydrolase (PAFAH). The data will be related to lipoprotein subclass distribution, and cross-sectionally and prospectively to vascular complication status, randomization groups, and interventions. 2. To define the roles of oxidative stress, inflammation, and homocysteine in the vascular complications of Type 1 and Type 2 diabetes. We hypothesize that oxidative stress, inflammatory processes, and elevated homocysteine levels mediate endothelial injury and accelerated atherosclerosis in diabetes. In both cohorts, we will determine products of free radical oxidation and antioxidant reserves in plasma to assess oxidative stress, C-reactive protein and serum amyloid A as measures of inflammatory processes, and plasma homocysteine levels.3. To determine the effect of lipoprotein associated enzymes on lipoprotein subclass distribution. We hypothesize that lipoprotein subclass distribution in serum is affected by LCAT, CETP, and PAFAH activity. Complementing studies of NMR determined lipoprotein subclass distribution and enzyme activities in vivo (Specific Aim 1); we will incubate serum with purified enzymes in vitro and determine effects on lipoprotein subclass distribution by NMR. 4. To determine functional characteristics of LDL, HDL, and their subclasses from Type 1 and Type 2 diabetic patients and control subjects in studies using human aortic endothelial cells (HAEC) and human red blood cell (RBC) membranes. We hypothesize that altered interactions of lipoproteins with vascular cells, related to dyslipidemia and oxidative stress, contribute to endothelial injury and accelerated atherosclerosis in diabetes. Using well- characterized lipoproteins from Type 1 and Type 2 diabetic subjects and from non-diabetic control subjects, we will determine: (i) effects of large, medium, and small LDL on HAEC expression of proteins mediating fibrinolysis, vascular tone, and adhesion molecules, and on matrix binding and intracellular calcium flux.(ii) ability of total HDL, and of large (cardioprotective) HDL2 and small (non-protective) HDL3 to:(a) mitigate oxidized-LDL induced HAEC adhesion molecule expression, tPA suppression, PAI-1 and NO production. (b) mitigate TNF- induced HAEC adhesion molecule expression(c) protect LDL and RBC membranes from in vitro oxidation(d) remove pre-formed lipid peroxides from oxidized RBC membranes

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们的主要目标是确定血脂异常和氧化应激在加速糖尿病大血管疾病（动脉粥样硬化）中的作用。 微血管疾病（肾病和视网膜病变）将作为次要终点进行研究。我们的项目在该计划的广泛范围中的位置如图1所示。 我们将继续（并扩大）对1型糖尿病患者DCCT/EDIC队列的纵向和横断面研究。该队列是世界上最大和最具特征的1型糖尿病患者组。 虽然还相对年轻，但它在方案项目的第一个供资期（1996年至今）提供了令人兴奋的数据。 为了获得更高的“硬”事件发生率（心肌梗死、卒中等）为了研究更常见的糖尿病形式，我们将从VA合作试验“2型糖尿病的血糖和并发症”中增加一个2型糖尿病队列。 这项前瞻性试验于2000年10月开始，与DCCT一样，患者将随机接受强化血糖控制和标准血糖控制。 VA试验为我们提供了在研究的随机化部分研究患者的机会（在DCCT中，随机化于1993年完成）。 我们已经获得了临时资金，以支持我们在第一年与VA试验的合作。 我们还将进行基础科学研究，以调查我们在过去四年中开发的与糖尿病血脂异常，氧化应激和动脉粥样硬化形成有关的假设。 这些研究与两项研究的其他项目和核心以及协调中心合作，将大大增加我们对两种类型糖尿病血管疾病背后复杂的多因素机制的了解。 我们的具体目标是：1。 明确血脂异常在1型和2型糖尿病血管并发症中的作用。我们将进行全面的脂蛋白分析，包括核磁共振（NMR）脂蛋白谱，载脂蛋白，脂蛋白（a）的水平，和脂蛋白相关酶的研究。 我们假设血脂异常，包括定量和定性脂蛋白异常，现在可以比以往任何时候都更详细地评估，介导内皮细胞损伤和激活，促进糖尿病动脉粥样硬化的加速。 我们将在两个场合获得DCCT/EDIC 1型糖尿病队列（“1型队列”）中选定受试者的NMR脂蛋白谱，在整个DCCT/EDIC研究期间（1983-2006）共完成7次测定。 我们还将在2000-2006年期间在VA合作试验2型糖尿病队列（“2型队列”）中两次前瞻性获得NMR分析。 我们将测量载脂蛋白A1、B、E和Lp（a）。 我们将测定以下脂蛋白相关酶的活性：对氧磷酶（PON）、卵磷脂胆固醇酰基转移酶（LCAT）和血小板活化因子乙酰水解酶（PAFAH）。数据将与脂蛋白亚类分布相关，并与血管并发症状态、随机分组和干预措施进行横断面和前瞻性相关。 2. 明确氧化应激、炎症和同型半胱氨酸在1型和2型糖尿病血管并发症中的作用。 我们推测氧化应激、炎症过程和同型半胱氨酸水平升高介导了糖尿病患者的内皮损伤和动脉粥样硬化的加速。 在这两个队列中，我们将测定血浆中自由基氧化产物和抗氧化剂储备，以评估氧化应激、C反应蛋白和血清淀粉样蛋白A作为炎症过程的指标，以及血浆同型半胱氨酸水平. 探讨脂蛋白相关酶对脂蛋白亚类分布的影响。 我们推测血清中脂蛋白亚类分布受LCAT、CETP和PAFAH活性的影响。 作为对NMR测定脂蛋白亚类分布和体内酶活性研究的补充（具体目标1）;我们将在体外用纯化酶孵育血清，并通过NMR测定对脂蛋白亚类分布的影响。4. 在使用人主动脉内皮细胞（HAEC）和人红细胞（RBC）膜的研究中，确定1型和2型糖尿病患者和对照受试者的LDL、HDL及其亚类的功能特征。 我们推测，脂蛋白与血管细胞的相互作用改变，与血脂异常和氧化应激有关，有助于糖尿病内皮损伤和加速动脉粥样硬化。 使用来自1型和2型糖尿病受试者以及来自非糖尿病对照受试者的充分表征的脂蛋白，我们将确定： 大、中、小LDL对HAEC介导纤维蛋白溶解、血管张力和粘附分子的蛋白质表达的影响，以及对基质结合和细胞内钙通量的影响。（二） 总HDL以及大（心脏保护性）HDL 2和小（非保护性）HDL 3的能力：（a） 减轻氧化LDL诱导的HAEC粘附分子表达、tPA抑制、派-1和NO产生。 （B） 减轻TNF-诱导的HAEC粘附分子表达（c）保护LDL和RBC膜免受体外氧化（d）从氧化的RBC膜中除去预先形成的脂质过氧化物