Nitric Oxide-Superoxide Interactions in Endothelial Cell Dysfunction

一氧化氮-超氧化物相互作用在内皮细胞功能障碍中的作用

基本信息

批准号：
8051607
负责人：
MAHENDRA KAVDIA
金额：
$ 34.62万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-14 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8051607
关键词：
Antioxidants Apoptosis Biological Availability Biological Process Blood Vessels Blood flow Cells Complex Computer Simulation Data Development Diabetes Mellitus Endothelial Cells Endothelium Environmental Risk Factor Enzymes Feedback Free Radical Formation Functional disorder Generations Glucose Goals Human Hyperglycemia Impairment In Vitro Kinetics Lipid Peroxidation Literature Measurement Measures Mediating Microcirculation Modeling Molecular Nitrates Nitric Oxide Nitric Oxide Synthase Nitrites Oxidative Stress Pathway interactions Peroxonitrite Process Production Reaction Reactive Nitrogen Species Reactive Oxygen Species Regulation Research Simulate Superoxide Dismutase Superoxides System Testing Time Tissues Umbilical vein Vasodilation abstracting cell injury design diabetes mellitus therapy diabetic human NOS3 protein improved multi-scale modeling prevent research study response shear stress therapy development

项目摘要

Abstract Endothelial cell (EC) dysfunction is a common pathogenic framework of many of the diabetes-related micro- and macro- vascular complications. Reduced bioavailability of EC-released nitric oxide (NO) is a primary marker generally used for EC dysfunction. However, the molecular mechanisms of hyperglycemia induced reduced NO bioavailability remain poorly understood. [We hypothesize that the diabetic endothelial cell dysfunction/reduced NO bioavailability is mediated by reactive oxygen species (ROS) and is a results of increased interaction of NO and superoxide (O2-) at the endothelial cell level. The increased interactions results in higher peroxynitrite (ONOO-) formation, shifting nitric oxide synthase (eNOS) activity from NO production to O2¿ production, and EC damage. The deleterious effects can be prevented by reducing ROS formation and concentration. Specific aims are designed to test these hypotheses. Aim1. Determine the EC release of NO and O2- and cell damage in hyperglycemic conditions. Hypotheses are: i) the high glucose causes endothelial dysfunction over long periods by increasing ONOO- and O2- formation and ii) reduction in O2- formation is key to reducing endothelial dysfunction. We will perform the following experiments: i) determining the effect of high glucose on eNOS and NA(D)PH expressions, NO and superoxide releases, endothelial cell lipid peroxidation (an indicator of peroxynitrite formation) and apoptosis in human umbilical vein endothelial cells (HUVECs) over short and long time-periods, and ii) determining whether increasing NO or decreasing O2- formation will be effective in preventing effects of high glucose. Aim2. Develop a reaction kinetic-transport computational model to simulate experiments of Aim1 and predict levels of NO, O2- and ONOO- at EC level. Hypotheses are: i) the NO concentration is reduced and ONOO- is increased due to high interaction between NO and O2- even though the NO release from endothelial cell increases in high glucose over short periods and ii) the NO concentration increases and ONOO- concentration decreases when O2- formation or concentration is reduced in high glucose. Aim3. Develop a multi-scale computational model for NO, ROS (O2-), and reactive nitrogen species (RNS; ONOO-) transport in the microcirculation underlying the process of oxidative stress. Hypotheses are: i) endothelial cell dysfunction is a results of higher superoxide formation, ii) a reduction in ROS formation enhances NO bioavailability and iii) increased superoxide dismutase levels not only reduces the O2- levels but also increases the NO levels, and reduces NO formation through feedback mechanism. At EC level, we will model the regulation of eNOS and the release of NO and O2-. At tissue level, we will model a volume of tissue containing an arteriolar blood vessel and simulate transport of NO, ROS and RNS.] This combined experimental & computational approach is critical in our understanding of molecular mechanism of EC dysfunction and examine the potential therapies to treat EC dysfunction related vascular complications. Project Narrative Endothelial cell (EC) dysfunction is a common pathogenic framework of many of the diabetes-related micro- and macro- vascular complications. The molecular mechanisms of hyperglycemia induced endothelial cell dysfunction remain poorly understood. The proposed research will use integrated computational and experimental approaches to assess endothelial cell dysfunction caused by oxidative stress due to high glucose at the molecular, cellular and tissue levels. The integrated experimental measurements and computational modeling of oxidative stress will provide an optimum set of parameters that will not only improve endothelial cell dysfunction/NO bioavailability but also will guide us in the development of therapies for diabetes related vascular complications.

抽象的内皮细胞（EC）功能障碍是许多与糖尿病相关的微型的常见致病框架和宏 - 血管并发症。 EC释放的一氧化氮（NO）的生物利用度降低是主要的标记通常用于EC功能障碍。但是，高血糖诱导的分子机制降低的生物利用度仍然很少了解。 [我们假设糖尿病内皮细胞功能障碍/降低没有生物利用度是由活性氧（ROS）介导的，是由在内皮细胞水平上NO和超氧化物（O2-）的相互作用增加。增加的相互作用结果在较高的过氧亚硝酸盐（ONOO-）形成中，一氧化氮合酶（ENOS）活性从无生产转变为 O2€生产和EC损害。可以通过减少ROS的形成和专注。具体目标旨在检验这些假设。 AIM1。确定否定的EC释放在高血糖条件下的O2和细胞损伤。假设是：i）高葡萄糖会导致内皮长时间通过增加ONOO-和O2形成和II）的功能障碍是关键的O2形成是关键减少内皮功能障碍。我们将执行以下实验：i）确定高葡萄糖在eNOS和Na（D）pH表达式上，NO和超氧化物释放，内皮细胞脂质过氧化（过氧亚硝酸盐形成的指标）和人脐静脉内皮细胞（HUVEC）的凋亡短期和长时间的周期，以及ii）确定NO不增加或减少O2形成将是有效预防高葡萄糖的作用。 AIM2。开发反应动力学传输计算模型为了模拟AIM1的实验，并在EC级别预测NO，O2-和ONOO-的水平。假设是：i）由于NO和O2之间的高度相互作用，均不降低浓度，并且在ONOO上增加了浓度 - 即使在短时间内，内皮细胞的无释放在高葡萄糖中增加，ii）无浓度当O2形成或浓度降低时，增加和浓度会下降葡萄糖。 AIM3。为NO，ROS（O2-）和反应性氮种开发多尺度计算模型（RN； onoo-）在微循环中的转运，氧化应激过程。假设是：i）内皮细胞功能障碍是较高的超氧化物形成的结果，ii）ROS形成的降低没有增强生物利用度，iii）升高的超氧化物歧化酶水平不仅会降低O2水平，而且还会降低还增加了NO水平，并通过反馈机制降低了NO形成。在EC级别，我们将建模eNOS的调节和NO和O2-的释放。在组织水平上，我们将建模一定数量的组织包含小动脉血管并模拟NO，ROS和RNS的运输。] 实验和计算方法对于我们对EC分子机制的理解至关重要功能障碍并检查治疗与功能障碍相关的血管并发症的潜在疗法。项目叙述内皮细胞（EC）功能障碍是许多与糖尿病相关的微型的常见致病框架和宏 - 血管并发症。高血糖诱导内皮细胞的分子机制功能障碍仍然知之甚少。拟议的研究将使用综合计算和评估由高葡萄糖引起的氧化应激引起的内皮细胞功能障碍的实验方法在分子，细胞和组织水平上。集成的实验测量和计算氧化应激的建模将提供一组最佳的参数，不仅可以改善内皮细胞功能障碍/无生物利用度，但也将指导我们开发与糖尿病有关的疗法血管并发症。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Induced peroxidase and cytoprotective enzyme expressions support adaptation of HUVECs to sustain subsequent H2O2 exposure.

诱导的过氧化物酶和细胞保护酶表达支持 HUVEC 的适应以维持随后的 H2O2 暴露。

DOI：
10.1016/j.mvr.2015.09.003
发表时间：
2016-01
期刊：
Microvascular research
影响因子：
3.1
作者：
Patel H;Chen J;Kavdia M
通讯作者：
Kavdia M

Modeling of biopterin-dependent pathways of eNOS for nitric oxide and superoxide production.

DOI：
10.1016/j.freeradbiomed.2011.06.009
发表时间：
2011-10-01
期刊：
Free radical biology & medicine
影响因子：
7.4
作者：
Kar S;Kavdia M
通讯作者：
Kavdia M

Computational insights into the role of glutathione in oxidative stress.

DOI：
10.2174/1567202611310020011
发表时间：
2013-05
期刊：
Current neurovascular research
影响因子：
2.1
作者：
Caitlin E Presnell;Gaurav Bhatti;L. Numan;M. Lerche;Salem K Alkhateeb;M. Ghalib;M. Shammaa;M. Kavdia-M.
通讯作者：
Caitlin E Presnell;Gaurav Bhatti;L. Numan;M. Lerche;Salem K Alkhateeb;M. Ghalib;M. Shammaa;M. Kavdia-M.

Mathematical and computational models of oxidative and nitrosative stress.

氧化和亚硝化应激的数学和计算模型。