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.

心血管疾病是老年人死亡的主要原因。 65岁的美国人，尤其是血管内皮细胞功能 显著下降，导致明显的血管僵硬 随着年龄的增长。氧化应激增加，细胞氧化还原改变，并改变 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，而这又可能是由线粒体衰变引起的。 然而，目前对线粒体的功能知之甚少。 血管内皮细胞，线粒体的范围或精确性质 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。该提案旨在填补以下方面的重大空白 血管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？，以及C)细胞的操纵 和/或添加抗坏血酸(AA)的线粒体抗氧化剂水平 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个月中调查这些问题 具体目标：1)比较与年龄相关的线粒体功能变化 新鲜分离的猪主动脉内皮细胞的氧化通量 (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(生物能量学、超氧化物生产和 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 细胞的抗氧化状态和氧化应激。2)评估以下项目的影响 PAEC功能的线粒体衰退。我们将把线粒体 腐烂，特别是在ROS增加和硫醇变化方面 氧化还原状态对PAEC一氧化氮合酶(ENOS)活性和 一氧化氮(EDNO)的可用性。3)确定操作是否 细胞抗氧化剂和硫醇氧化还原状态的研究 AA或LA在体外(PAEC)或体内(豚鼠主动脉环)的添加 改善线粒体功能和内皮依赖性生物 活动。AA或LA将添加到新分离的PAEC中 和老猪来确定这些制剂是反转还是“伪装” 线粒体依赖于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 将通过补充AA和AA豚鼠喂养来增加 以确定线粒体功能(LA)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 活体内与年龄相关的血管张力丧失。