PURINERGIC AXIS OF CARDIAC BLOOD VESSELS

心脏血管的嘌呤能轴

• 批准号: 2609377
• 负责人: IAIN L BUXTON
• 金额: $ 23.48万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-12-01 至 2001-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2609377
• 关键词: adenosine triphosphate; adenylate kinase; coronary vessels; guinea pigs; hormone regulation /control mechanism; hypoxia; nitric oxide; nucleoside phosphate kinase; nucleotide metabolism; prostacyclins; proteolysis; purinergic receptor; reperfusion; tissue /cell culture; vascular endothelium; vasomotion; western blottings

The goal of the proposed research is to explain the role of extracellular adenine nucleotides, particularly adenosine 5'- triphosphate (ATP), in normal cardiac endothelial cell (EC) function where release of factors that participate in the regulation of blood flow may be important in normal cardiac function and in cardiovascular disease. ECs have been shown to release ATP and autacoids such as endothelium-dependent relaxing factor and prostacyclin in response to various stimuli, including ATP itself. In addition, we propose that ECs can convert adenosine 5'-diphosphate (ADP) to ATP extracellularly to maintain the presence of ATP-mediated relaxation of the coronary and resistance vessels down-stream from the site of ATP release. We suggest that control of ATP appearance outside the EC and its eventual conversion to adenosine is an important component of regional blood flow control in the normal heart. As it is not clear how release of ATP or conversion of ADP to ATP by ECs is regulated, specific aims have been designed as follows: 1. Test the hypothesis that the release of ATP from ECs and the conversion of ADP to ATP by ECs can be measured in an intact coronary artery and that ATP release can support nucleoside triphosphate formation. Furthermore, to test the hypothesis that conversion of ADP to ATP can be inhibited in situ and determine the affect of hypoxia on extracellular generation of ATP. 2. Test the hypothesis that cardiac EC release of ATP in response to agonists occurs both luminally and abluminally and test the effect of hypoxia and re-oxygenation on this process. 3. Explore the manner in which ECs release ATP and test the hypothesis that the elaboration of ATP by stimulated ECs is associated, mechanistically, with the ability of ECs to release nitric oxide. 4. Assess the role of ecto-nucleoside diphosphate kinase and ecto- adenylate kinase in the extracellular production of ATP by ECs and test effects of hypoxia and reoxygenation on the activity of these enzymes. We will approach these aims with biochemical, pharmacological, molecular, cell biological and microscopic methods employing intact blood vessels, cells in primary culture and subcellular fractions.

拟议研究的目的是解释 细胞外腺嘌呤核苷酸，特别是腺苷5'-- 三磷酸（ATP），正常心脏内皮细胞（EC）功能 释放参与血液调节的因素 流动在正常心脏功能和心血管中可能很重要 疾病。 EC已显示出释放ATP和自闭症（例如 内皮依赖性放松因子和前列环蛋白响应于 各种刺激，包括ATP本身。此外，我们建议ECS 可以将5'-二磷酸腺苷（ADP）转换为ATP的细胞外 保持ATP介导的冠状动脉放松的存在 电阻容器从ATP释放位置向下进行。我们建议 控制EC之外的ATP外观及其最终的控制 转换为腺苷是区域血的重要组成部分 正常心脏的流动控制。因为尚不清楚ATP的释放如何 或受ECS将ADP转换为ATP，具体目的已有 被设计如下： 1。检验以下假设：ATP从EC和 可以在完整的冠状动脉中测量ADP向ATP转换为ATP 动脉和ATP释放可以支持三磷酸核苷 形成。此外，要检验ADP转化的假设 可以原位抑制ATP，并确定缺氧的影响 细胞外产生ATP。 2。检验以下假设：响应于ATP的心脏EC释放 激动剂既有透明地和空白的发生，并测试 在此过程中缺氧和重新融合。 3。探索ECS释放ATP的方式并检验假设 通过刺激EC阐述ATP是相关的， 机械上，EC释放一氧化氮的能力。 4。评估核核苷双磷酸激酶和肾小管的作用 EC的ATP细胞外产生中的腺苷酸激酶和测试 缺氧和重氧对这些酶活性的影响。 我们将通过生化，药理学， 使用完整的分子，细胞生物学和微观方法 血管，原发性培养物中的细胞和亚细胞级分。