Vascular Reactivity: Gender and Hormonal Influence

血管反应性：性别和激素的影响

基本信息

批准号：
7576721
负责人：
SUE P DUCKLES
金额：
$ 38.13万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-07-01 至 2012-03-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7576721
关键词：
Aconitate Hydratase Address Age Aging Agonist Antibodies Antioxidants Biogenesis Biological Assay Blood Vessels Brain Cells Cerebrum DNA Disease Down-Regulation Dyes Elderly Electron Transport Endothelial Cells Enzymes Equilibrium Estrogen Receptor alpha Estrogen Receptor beta Estrogens Exhibits Exposure to Family Female Fumarate Hydratase Functional disorder Gender Glucose Harvest Hormonal Human Immunoblotting Immunoprecipitation In Vitro Iron Ischemia Knowledge Measures Membrane Potentials Messenger RNA Mitochondria Mitochondrial DNA Mitochondrial Proteins Modeling Mus Nitric Oxide Synthase Nuclear Oxidative Phosphorylation Oxidative Stress Oxygen Process Production Proteins Reactive Oxygen Species Regulation Research Personnel Respiration Reverse Transcriptase Polymerase Chain Reaction Role Small Interfering RNA Stress Stroke Sulfur Superoxides Testing Transcription Coactivator Transgenic Mice Vascular Dementia Vascular Diseases Vascular System age related aged cerebral artery cerebrovascular deprivation diet and exercise enzyme activity improved in vivo in vivo Model indexing inhibitor/antagonist insight middle cerebral artery mitochondrial dysfunction mitochondrial membrane mouse model mtTF1 transcription factor novel nrf1 protein nuclear respiratory factor oxidative damage prevent programs receptor research study response transcription factor

项目摘要

DESCRIPTION (provided by applicant): Mitochondrial dysfunction has been implicated as a cause of age-related disorders; however little is known regarding the role of mitochondria in vascular disease. We recently found that estrogen (E) has beneficial effects on cerebrovascular mitochondria that likely contribute to vasoprotection. Thus, our overall hypothesis is: Effects of E on mitochondrial function protect against cerebrovascular endothelial dysfunction with age. In this proposal, mechanisms and consequences of E modulation of cerebrovascular mitochondria will be determined both in vitro and in vivo. Cultured human brain endothelial cells will be used in the first two aims. HYPOTHESIS 1: E increases the efficiency of oxidative phosphorylation and decreases oxidative stress in cerebral endothelial mitochondria. Key indices of mitochondrial function, mitochondrial enzyme levels and activities, mitochondrial biogenesis, and production of reactive oxygen species (ROS) will be measured after E treatment in normal conditions and in culture models of ischemia. The involvement of E receptors (ERalpha and ERbeta) will be tested using selective siRNA knockdown, antagonists, agonists and selective ER modulators (SERMs). HYPOTHESIS 2: E regulates mitochondrial function via PGC-1 transcriptional coactivators. We will determine effects of E, and the role of ERs, on levels of mitochondria-related transcription factors, including the PGC-1 family. Specific involvement of PGC-1¿ in mitochondrial effects of E will be tested using siRNA downregulation of PGC-1¿. ER-PGC-1¿ interactions will be tested by immunoprecipitation. In aim 3, a transgenic mouse model (MnSOD) will be used to test HYPOTHESIS 3: In vivo E protects cerebrovascular mitochondria and decreases endothelial dysfunction with advancing age. Cerebral blood vessels will be isolated from intact, ovariectomized, and E-treated females of different ages to compare mitochondrial enzymes, ATP and ROS production, and damage to mitochondrial iron/sulphur- containing enzymes and mtDNA. Endothelial function will be assessed in pressurized segments of isolated middle cerebral arteries. This project will delineate novel ways in which estrogen and estrogen-related therapies modulate the cerebral vascular system. These studies may provide insight into the role of mitochondria in cerebrovascular dysfunction, a contributor to age-related disorders such as stroke and vascular dementia.

描述（由申请人提供）：线粒体功能障碍被认为是与年龄相关的疾病的一个原因；然而，我们最近发现雌激素（E）对脑血管线粒体可能具有有益作用。因此，我们的总体假设是：E 对线粒体功能的影响可防止随年龄增长而出现的脑血管内皮功能障碍。脑血管线粒体将在体外和体内进行测定。假设 1：E 提高脑内皮线粒体功能的氧化应激效率。在正常条件下和缺血培养模型中，E处理后将测量线粒体酶水平和活性、线粒体生物合成和活性氧（ROS）的产生。 E 受体（ERα 和 ERβ）的参与将使用选择性 siRNA 敲低、拮抗剂、激动剂和选择性 ER 调节剂 (SERM) 进行测试。假设 2：E 通过 PGC-1 转录共激活因子调节线粒体功能。 ER 对线粒体相关转录因子（包括 PGC-1 家族）水平的作用。 E 在线粒体中的作用将使用 PGC-1 的 siRNA 下调进行测试¿ ER-PGC-1¿ 在目标 3 中，将使用转基因小鼠模型 (MnSOD) 来测试假设 3：体内 E 可以保护脑血管线粒体并随着年龄的增长而减少内皮功能障碍。和 E 处理的不同年龄的女性，比较线粒体酶、ATP 和 ROS 的产生，以及对线粒体含铁/硫酶的损害和mtDNA。该项目将在孤立的大脑中动脉的加压部分中评估内皮功能，这些研究可能会深入了解线粒体在脑血管功能障碍中的作用。导致中风和血管性痴呆等与年龄相关的疾病。