Vitamin C, glutathione and mitochondrial function

维生素 C、谷胱甘肽和线粒体功能

• 批准号: 6369053
• 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: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6369053
• 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.

心血管疾病是人们的主要死亡原因 尤其是美国的65岁，血管内皮细胞功能 明显下降，促成了明显的船只僵化 随着年龄的增长。增加氧化应激，细胞氧化还原变化和改变 钙稳态可能是与年龄有关的内皮的潜在因素 细胞功能障碍反过来可能是由线粒体衰减引起的。 但是，关于线粒体功能在 血管内皮细胞，线粒体的程度或精确性 随着年龄的增长或这种衰减对内皮细胞的重要性 功能。该建议旨在填补这些重大空白 内皮细胞随着年龄的增长吗？ b）线粒体衰减会影响内皮 细胞功能和容器音调？和c）操纵细胞是否会操纵 和/或线粒体抗氧化剂水平通过添加抗坏血酸（AA） 或氟酸（LA）反向线粒体衰减的后果 内皮功能？我们建议在3中调查这些问题 具体目的：1）比较线粒体功能的年龄相关变化 新鲜分离的猪主动脉内皮细胞中的氧化剂通量 （PAEC）来自年轻猪和老猪。这些研究将定义衰老 影响线粒体功能（生物能学，超氧化物产生和 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 细胞抗氧化剂状态和氧化应激。 2）评估 PAEC功能的线粒体衰减。我们将关联线粒体 衰减，特别是就ROS增加和硫醇改变而言 氧化还原状态，对一氧化氮合酶（ENOS）活性和 一氧化氮（EDNO）的可用性。 3）确定是否操纵 细胞抗氧化剂和硫醇氧化还原状态的细胞抗氧化剂 在体外（PAEC）或体内添加AA或LA（豚鼠主动脉环） 改善线粒体功能和内皮依赖性生物学 活动。 AA或LA将添加到来自年轻的新鲜隔离的PAEC中 和旧猪来确定这些药物是逆转还是“掩盖” 线粒体依赖于细胞氧化还原状态的改变，氧化 压力（AIM 1）和内皮生物学活性（AIM 2）。这些研究 通过补充豚鼠和AA和 LA确定线粒体功能是否增加（LA）和/或 抗氧化剂和硫醇氧化还原状态（AA或LA）的增加可能会逆转 体内血管张力中与年龄相关的损失。