Protein S-glutathionylation and vascular dysfunction with aging

衰老过程中蛋白质 S-谷胱甘肽化和血管功能障碍

• 批准号: 9751680
• 负责人: JINGYAN HAN
• 金额: $ 21.11万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751680
• 关键词: Acetylcholine; Adult; Aging; Alzheimer' s Disease; American; Antioxidants; Aorta; Arteries; Biological Availability; Blood Vessels; Cardiac; Cardiovascular Diseases; Cardiovascular Physiology; Cell physiology; Cells; Cysteine; Data; Down-Regulation; Doxycycline; Elderly; Endothelial Cells; Endothelin-1; Endothelin-2; Endothelium; Etiology; Experimental Models; Fatty Liver; Functional disorder; Gene Proteins; Grant; Grx1 protein; Health; Hemostatic function; Human; Hyperlipidemia; Imaging Techniques; Impairment; Knockout Mice; Knowledge; Lead; Lipid Peroxidation; Literature; Longevity; Measures; Mediating; Metabolic; Metabolic syndrome; Mission; Modeling; Modification; Molecular; Mus; Nitrates; Obesity; Older Population; Optical Coherence Tomography; Oxidants; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Pathogenicity; Patients; Play; Population; Post-Translational Protein Processing; Predisposition; Process; Production; Protein S; Proteins; Proteomics; Public Health; Publishing; Reactive Oxygen Species; Regulation; Relaxation; Research; Role; SIRT1 gene; Signal Pathway; Signal Transduction; Signaling Molecule; Sulfhydryl Compounds; Symptoms; Techniques; Testing; Tetanus Helper Peptide; Therapeutic; Transgenic Mice; Tyrosine; United States National Institutes of Health; Up-Regulation; Vascular Diseases; Vasoconstrictor Agents; Vasodilation; Vasodilator Agents; Work; age effect; age related; aged; aging brain; antioxidant therapy; cardiovascular risk factor; catalyst; cell growth regulation; driving force; endothelial dysfunction; femoral artery; glutaredoxin; human old age (65+); hypercholesterolemia; in vivo; loss of function; middle age; mouse model; nitration; novel; novel strategies; older patient; overexpression; oxidative damage; preservation; prevent; response; targeted treatment; transgene expression; vascular endothelial dysfunction

PROJECT SUMMARY Vascular endothelial dysfunction (VED) is a key feature of vascular aging underlying the predisposition of old adults to CVD. Sustained oxidative stress is a major driving force for VED by synergically dysregulating the production of vasodilator NO and vasoconstrictor endothelin-1(ET-1). But the underlying redox mechanisms re- main unclear. Novel approaches to reversing oxidative stress are particularly compelling given the ineffective- ness of general antioxidant therapies that targets extremely short-lived ROS. Protein S-glutathionylation (PrS- SG), a stable but reversible oxidant-induced posttranslational modification dominates the molecular mechanisms for redox signaling. Reversal of PrS-SG is catalyzed specifically by glutaredoxin which thereby is critical in redox regulation of cellular function. Dysfunction of glutaredoxin-1 (Glrx-1), a major actor in the de-glutathionylation is implicated in cardiac, and brain aging. Its role in vascular aging is unknown. This application is built upon a strong premise. Our published and preliminary studies indicate that Glrx1 deficiency and PrS-SG induction promote endothelial dysfunction and aging-associated metabolic syndromes and that during aging, aortic Glrx1 is decreased with a concomitant increase in endothelial PrS-SG. Evidence in the literature also supports a causal role of Pr-SSG in VED in that S-glutathionylation can uncouple eNOS and activate p21Ras, which are known to impair NO bioavailability and stimulate Erk1/2 dependent-ET-1 expression. We thus hypothesize that age-related Glrx1 downregulation promotes VED by dysregulating the integrated redox signaling of eNOS and ET-1, which can be reversed by replenishing Grlx1 in endothelium. This central hypothesis is tested by pursuing two specific aims: 1) To understand the relationship between Glrx1/PrS-SG and the onset and progression of VED with aging. Using C57BL6J mice, a well-characterized model of aging we will measure the temporal changes in aortic Glrx1, PrS-SG, and endothelium-dependent vasorelaxation in young, middle-aged and old mice, and longitudinal changes of flow-mediated dilation of femoral artery in vivo using a non-invasive Optical-Coherence-Tomography technique. To test the causal role of Glrx1 downregulation in VED, we will test whether VED is aggravated in Glrx1 knockout mice; 2) To test a new concept that replenishing Glrx1 in endothelium can reverse VED in aging. Using a novel inducible endothelial specific Glrx1 transgenic mouse model, we will determine the impact on VED and the integrated signaling of eNOS and ET-1 of Glrx1 transgene expression in young, middle-aged, and old mice. The positive results in the R21 grant will help advance our understanding how redox signaling mediates the multifaceted effects of aging on ECs, promoting VED, and offer a new antioxidant therapeutic strategy to restore vascular function in older adults.

项目摘要 血管内皮功能障碍（VED）是旧血管衰老的关键特征 成人到CVD。持续的氧化应激是通过协同失调的VED的主要驱动力 血管扩张剂NO和血管收缩器内皮素-1（ET-1）的产生。但是基本的氧化还原机制恢复了 主要不清楚。鉴于无效 - 靶向极短的ROS的一般抗氧化剂疗法的NES。蛋白S-谷胱甘肽化（PRS- SG），一种稳定但可逆的氧化剂诱导的翻译后修饰，主导了分子机制 用于氧化还原信号。 PRS-SG的逆转是由谷胱甘肽专门催化的，因此在氧化还原中至关重要 调节细胞功能。谷胱甘肽-1（GLRX-1）的功能障碍，De-Glutathionylation中的主要参与者是 与心脏和大脑衰老有关。它在血管衰老中的作用尚不清楚。该应用是建立在强大的基础上的 前提。我们发表的初步研究表明，GLRX1缺乏症和PRS-SG诱导促进 内皮功能障碍和与衰老相关的代谢综合症，并且在衰老期间，主动脉GLRX1为 随着内皮PRS-SG的同时增加而减少。文献中的证据也支持因果关系 PR-SSG在S-glutathionylation中的VED中的作用可以取消eNOS并激活P21RA，这是已知的p21RA 损害没有生物利用度并刺激ERK1/2依赖性-ET-1表达。因此，我们假设与年龄有关 GLRX1下调通过失调的ENOS和ET-1的综合氧化还原信号传导来促进VED， 可以通过在内皮中补充GRLX1来逆转。通过追求两个特定的特定假设来检验该中心假设 目的：1）了解GLRX1/PRS-SG与衰老的VED的发作和进展之间的关系。 使用C57BL6J小鼠（一种陈化的良好模型），我们将测量主动脉glrx1的时间变化， PRS-SG和年轻，中年和老鼠的依赖于内皮的血管征为纵向 使用非侵入性光学 - 造影术在体内流动介导的股动脉扩张的变化 技术。为了测试GLRX1下调在VED中的因果作用，我们将测试VED是否在 GLRX1敲除小鼠； 2）测试一个新概念，以补充内皮中的GLRX1可以逆转衰老。 使用新型的诱导内皮特异性GLRX1转基因小鼠模型，我们将确定对VED的影响 以及年轻，中年和年龄的GLRX1转基因表达的eNOS和ET-1的集成信号传导 老鼠。 R21赠款的积极结果将有助于我们理解氧化还原信号如何介导 衰老对EC的多方面影响，促进VED，并提供一种新的抗氧化治疗策略 老年人恢复血管功能。