Ethanol Prevents Microvascular Dysfunction

乙醇预防微血管功能障碍

• 批准号: 7036114
• 负责人: RONALD JOHN KORTHUIS
• 金额: $ 33.33万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-15 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7036114
• 关键词: alcoholic beverage consumption; antioxidants; biological signal transduction; calcitonin gene related peptide; cardiovascular disorder prevention; cell adhesion; cell cell interaction; chloride channels; cytochrome P450; enzyme activity; enzyme complex; enzyme induction /repression; ethanol; gene deletion mutation; genetically modified animals; inflammation; intravital microscopy; laboratory mouse; microcirculation; monoclonal antibody; nitric oxide; nitric oxide synthase; phenotype; protein localization; selectins; serine threonine protein kinase; vascular endothelium

DESCRIPTION (provided by applicant): Ethanol consumption at low to moderate levels protects the heart and vasculature from the deleterious effects of ischemia and reperfusion (I/R). Our recent work indicates that antecedent ethanol ingestion provokes the development of an anti-inflammatory phenotype in postcapillary venules such that these microvessels fail to support leukocyte/endothelial cell adhesive interactions during I/R. Surprisingly, AMP-activated kinase (AMPK) plays a necessary role in initiating this adaptive transformation to a protected phenotype in postcapillary venules. However, it is not clear how this enzyme, best known for its role as a metabolic master switch, is involved in transducing signals elicited by ethanol to effect the acquisition of tolerance to I/R in the microcirculation. Our overall hypothesis is that ethanol-induced, AMPK-dependent eNOS activation serves as an important initiating factor to trigger a cascade of signaling events that ultimately act to promote the expression of cytochrome P450 epoxygenases (CYP), which serve as an important effector of the beneficial microvascular actions of ethanol by catalyzing the formation of reaction products which exhibit powerful anti-adhesive and antioxidant properties. To address this postulate, we propose to determine whether: (1) ethanol-induced AMPK activation initiates the development of an anti-inflammatory phenotype by stimulating the formation of eNOS-derived NO; (2) NO-induced release of calcitonin gene-related peptide (CGRP) plays an essential role in the acquisition of tolerance to ischemia that is evoked by antecedent ethanol consumption; (3) the cystic fibrosis transmembrane regulator (CFTR) serves as an obligatory downstream signaling element that participates in inaugurating the development of the protected phenotype precipitated by ethanol-induced AMPK activation; and (4) ethanol-induced, AMPK-triggered increases in CYP activity during I/R prevents postischemic leukocyte adhesion in postcapillary venules by abrogating P-selectin expression. Intravital microscopic approaches will be used to quantify leukocyte rolling and adhesion in wild-type control mice and in mutant mice lacking AMPK, eNOS, CGRP, or CFTR. The influence of ethanol on l/R-induced adhesion molecule expression, AMPK and eNOS activity, CGRP release, and CYP protein expression and activity will also be investigated. Collectively, these aims address novel mechanisms whereby antecedent ethanol ingestion induces the development of an anti-inflammatory phenotype in postcapillary venules. This work will identify new links between ethanol-induced, AMPK-dependent, eNOS-derived NO formation as essential triggering elements, CGRP release and CFTR function as obligatory downstream mediators, and increased CYP activity as a major effector in the acquisition of tolerance to I/R by antecedent ethanol ingestion.

描述（由申请人提供）：低至中等水平的乙醇消耗可保护心脏和脉管系统免受缺血和再灌注（I/R）的有害影响。我们最近的工作表明，先前的乙醇摄入引起了后毛细血管静脉静脉表型的发展，因此这些微血管在I/R期间无法支持白细胞/内皮细胞粘附相互作用。令人惊讶的是，AMP激活的激酶（AMPK）在启动这种适应性转化对后毛细血管静脉的适应性转化中起着必要的作用。但是，目前尚不清楚该酶是如何以代谢主开关的作用而闻名的，它参与了乙醇引起的传输信号，以实现微循环中对I/R的耐受性。我们的总体假设是，乙醇诱导的AMPK依赖性ENOS激活是触发一系列信号事件的重要启动因素，这些因素最终促进了促进细胞色素p450磷酸酶（CYP）的表达，它是通过猫化作品的强大估算的估算的重要效果，该作用是猫科动物的重要效果，并表现出鸡尾酒的形式，表现出了估算的形式，表现出了形式的形式，即形成型的形式，以表现出形式形式。 特性。为了解决这一假设，我们建议确定：（1）乙醇诱导的AMPK激活是否通过刺激eNOS衍生的NO的形成来启动抗炎表型的发展； （2）无诱导的降钙素基因相关肽（CGRP）在获取对缺血的耐受性中起着至关重要的作用，而耐血性乙醇的消耗引起的耐受性； （3）囊性纤维化跨膜调节剂（CFTR）是一种强制性的下游信号传导元件，参与了由乙醇诱导的AMPK激活沉淀的受保护表型的发展的启动； （4）I/R期间乙醇诱导的，AMPK触发的CYP活性增加，可通过废除p-选择素的表达来防止后毛细血管后毛细血管静脉内粘附。插入式显微镜方法将用于量化野生型对照小鼠的白细胞滚动和粘附，以及缺乏AMPK，ENOS，CGRP或CFTR的突变小鼠中。还将研究乙醇对L/R诱导的粘附分子表达，AMPK和ENOS活性，CGRP释放以及CYP蛋白表达和活性的影响。总的来说，这些目的涉及新的机制，从而诱导乙醇摄入的前乙醇在后毛细血管后静脉静脉静脉表型的发展。这项工作将确定乙醇诱导的，依赖AMPK的，eNOS衍生的NO形成之间的新联系，因为必不可少的触发元素，CGRP释放和CFTR功能作为强制性的下游介体，以及CYP的CYP活性增加，作为在前乙醇中获得I/R对I/R的耐受性的主要效果。