Vitamin C, glutathione and mitochondrial function

维生素 C、谷胱甘肽和线粒体功能

• 批准号: 6658446
• 负责人: TORY M HAGEN
• 金额: $ 26.22万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6658446
• 关键词: age difference; aging; ascorbate; bioenergetics; calcium flux; cell senescence; cellular respiration; dietary supplements; free radical oxygen; glutathione; guinea pigs; mature animal; mitochondria; nitric oxide; nitric oxide synthase; nutrition related tag; oxidative stress; short chain fatty acid; swine; vascular endothelium; vasomotion

Cardiovascular diseases are the major cause of death for people over the age of 65 in the U.S. In particular, vascular endothelial cell function markedly declines, contributing to the profound vessel rigidity evident with age. Increased oxidative stress, cellular redox changes, and altered calcium homeostasis may be underlying factors in age-related endothelial cell dysfunction, which in turn, could be caused by mitochondrial decay. However, there is nothing known about mitochondrial function in vascular endothelial cells, the extent or precise nature of mitochondrial decay with age, or the significance of such decay on endothelial cell function. This proposal is intended to fill these significant gaps in endothelial cells with age? B) does mitochondrial decay affect endothelial cell function and vessel tone?, and C) does manipulation of cellular and/or mitochondrial antioxidant levels by addition of ascorbic acid (AA) or lipoic acid (LA) reverse the consequences of mitochondrial decay to endothelial function? We propose to investigate these questions in 3 specific aims: 1) Compare age-related changes in mitochondrial function and oxidant flux in freshly isolated porcine aortic endothelial cells (PAEC) from young and old pigs. These studies will define how aging affects mitochondrial function (bioenergetics, superoxide production and calcium homeostasis) in PAEC. In particular, we will test how mitochondrial function derived reactive oxygen species (ROS) alter cellular antioxidant status and oxidative stress. 2) Assess the impact of mitochondrial decay of PAEC function. We will correlate mitochondrial decay, especially in terms in terms of increased ROS and altered thiol redox status, to PAEC nitric oxide synthase (eNOS) activity and availability of nitric oxide (EDNO). 3) Determine whether manipulation of cellular antioxidant of cellular antioxidant and thiol redox status by addition of AA or LA in vitro (PAEC) or in vivo (guinea pig aortic rings) improves mitochondrial function and endothelial-dependent biological activity. AA or LA will be added to freshly isolated PAEC from young and old pigs to determine whether these agents reverse or "mask" mitochondrial-dependent alterations to cellular redox status, oxidative stress (Aim 1) and endothelial biological activity (Aim 2). These studies will be augmented by supplemental feeding of guinea pigs with AA and LA to determine whether increased mitochondrial function (LA) and/or increased antioxidant and thiol redox status (AA or LA) may reverse the age-related loss in vascular tone in vivo.

心血管疾病是近年来人们死亡的主要原因 美国65岁特别是血管内皮细胞功能 显着下降，导致明显的血管刚性 随着年龄的增长。氧化应激增加、细胞氧化还原变化和改变 钙稳态可能是年龄相关内皮细胞的潜在因素 细胞功能障碍可能是由线粒体衰变引起的。 然而，对于线粒体的功能尚无所知。 血管内皮细胞，线粒体的程度或精确性质 随着年龄的增长而衰退，或这种衰退对内皮细胞的意义 功能。该提案旨在填补这些重大空白 内皮细胞随年龄增长？ B) 线粒体衰变是否影响内皮细胞 细胞功能和血管张力？，C）细胞的操纵 和/或通过添加抗坏血酸 (AA) 来提高线粒体抗氧化水平 或硫辛酸 (LA) 逆转线粒体衰变的后果 内皮功能？我们建议在 3 年内调查这些问题 具体目标：1）比较线粒体功能与年龄相关的变化 新鲜分离的猪主动脉内皮细胞中的氧化通量和氧化剂通量 (PAEC) 来自幼猪和老猪。这些研究将定义衰老如何 影响线粒体功能（生物能学、超氧化物产生和 PAEC 中的钙稳态）。特别是，我们将测试如何 线粒体功能衍生的活性氧 (ROS) 改变 细胞抗氧化状态和氧化应激。 2) 评估影响 PAEC 功能的线粒体衰退。我们将关联线粒体 衰变，特别是在活性氧增加和硫醇改变方面 氧化还原状态，PAEC 一氧化氮合酶 (eNOS) 活性和 一氧化氮 (EDNO) 的可用性。 3）判断是否被操纵 细胞抗氧化剂的细胞抗氧化剂和硫醇氧化还原状态 体外 (PAEC) 或体内（豚鼠主动脉环）添加 AA 或 LA 改善线粒体功能和内皮依赖性生物 活动。 AA 或 LA 将添加到新鲜分离的幼仔 PAEC 中 和老猪以确定这些药物是否逆转或“掩盖” 线粒体依赖性细胞氧化还原状态的改变，氧化 应激（目标 1）和内皮生物活性（目标 2）。这些研究 将通过用 AA 和 AA 补充喂养豚鼠来增强 LA 以确定线粒体功能 (LA) 是否增加和/或 增加抗氧化剂和硫醇氧化还原状态（AA 或 LA）可能会逆转 体内血管张力与年龄相关的损失。