CEREBROVASCULAR CONTRACTILE RESPONSES TO HIGH ALTITUDE LONG TERM HYPOXIA

高原长期缺氧的脑血管收缩反应

• 批准号: 8011799
• 负责人: LAWRENCE D LONGO
• 金额: $ 20.91万
• 依托单位: LOMA LINDA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8011799
• 关键词: Acclimatization; Acute; Address; Adrenergic Agents; Adrenergic Receptor; Adult; Agonist; Altitude; Altitude Sickness; Animals; Arteries; Attenuated; Biochemical; Biogenic Amines; Biological Assay; Blood Vessels; Blood flow; Brain; Brain Hypoxia-Ischemia; Calcium; Cardiovascular Diseases; Cations; Cell membrane; Cells; Cellular Retinol Binding Protein; Cerebral Edema; Cerebral Ischemia-Hypoxia; Cerebral hemisphere hemorrhage; Cerebrovascular Circulation; Cerebrum; Computer software; Confocal Microscopy; Coupling; Data Analyses; Dependence; Development; Disease; Domestic Sheep; Double-Stranded RNA; Elements; Endoplasmic Reticulum; Enzymes; Extracellular Signal Regulated Kinases; FOS gene; Fetus; Figs - dietary; Filament; Gel; Gene Activation; Gene Expression; Gene Expression Regulation; Gene Proteins; Genes; Genetic Transcription; Genomics; Growth and Development function; Heart; Hemorrhage; Homeostasis; Hypoxemia; Hypoxia; Image; Infant; Inositol; Intraventricular; Isoenzymes; JUN gene; Life; Light; Long-Term Effects; Lung diseases; MAPK1 gene; Maintenance; Maps; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Microfilaments; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Molecular; Molecular Profiling; Myosin ATPase; Myosin Heavy Chains; Neurologic; Newborn Infant; Nuclear Antigens; Organism; Pathway Analysis; Pathway interactions; Peptide Mapping; Peptides; Perfusion; Perinatal subependymal hemorrhage; Personal Satisfaction; Phenotype; Phenylephrine; Phosphorylation; Phosphotransferases; Physiological; Play; Potassium; Pre-Eclampsia; Predisposition; Protein Isoforms; Protein Kinase C; Proteins; Proteomics; Proto-Oncogenes; Pulmonary Edema; Pulmonary Hypertension; Pump; Qi; RNA; RNA Interference; Regulation; Research; Retinol Binding Proteins; Reverse Transcriptase Polymerase Chain Reaction; Rho-associated kinase; Role; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; Sarcoplasmic Reticulum; Sea; Second Messenger Systems; Secondary to; Sheep; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Signaling Pathway Gene; Signaling Protein; Smoke; Smooth Muscle Myocytes; Smooth Muscle Myosins; Solutions; Staining method; Stains; Stimulus; Stream; Stress; Testing; Thick; Transcript; Up-Regulation; Vascular Smooth Muscle; Western Blotting; Woman; Work; adrenergic; age difference; base; caldesmon; calponin; cell growth; cerebral artery; cerebrovascular; citrate carrier; extracellular; fetal; inhibitor/antagonist; insight; mRNA Expression; mature animal; microchip; multidisciplinary; novel; pregnant; programs; protein activation; protein expression; protein protein interaction; receptor; response; rho; second messenger; virtual

The overall theme of this project is to understand whereby acclimatization to high altitude long-term hypoxia (LTH) alters fundamental mechanisms in the cerebrovasculature of the fetus and adult. We also will examine these mechanisms in association with development from fetus to adult. This project is broadly based, multidisciplinary, and vertically integrated using physiologic, cellular, biochemical, and molecular approaches. Based on two decades of research, we shall test a number of hypotheses in sheep acclimatized to high altitude. The overall hypothesis is that high altitude, long-term hypoxia is associated with changes in cerebrovascular contractile responses secondary to altered alphai-adrenergic-receptor (or AR) subtypes and/or specific protein kinase C isoforms (PKC)-mediated downstream Ca2+-dependent and Ca -independent signal transduction pathways. An associated hypothesis is that LTH significantly alters Oi-AR-subtype- and specific PKC isozyme-mediated expression of proto-oncogenes and genes representing vascular smooth muscle "synthetic" and/or "proliferative" phenotypes, as compared to those of adult "contractile" phenotype. Four specific aims will examine the role of LTH in cerebral artery Or adrenergic-mediated signal tranduction of thin and thick myofilament, e.g., 1) roles of Oi-AR, PKC isoforms, extracellular signal regulated kinases (ERKs), and downstream effector proteins, 2) the role of plasma membrane and sarcoplasmic reticulum Ca2+ channels, 3) gene regulation of vascular phenotypes, and 4) signaling pathways gene/protein discovery. For the studies we will utilize agonist-induced contractility and intracellular [Ca2+], Western immunobiots, RT-PCR, RNAi silencing, confocal microscopy, 2D-gel-mass spectroscopy, and gene microarray/pathways analysis. Scientifically the studies will augment our understanding of basic mechanisms whereby fetal and adult cerebral vessels acclimatize to LTH. They also will illustrate aspects of development from fetus to adult. Clinically the studies relate to at least three critical problems. 1) For the fetus and newborn they relate to responses to prolonged hypoxia as occurs in women who live at high altitude, as well as those who smoke or are anemic, who have heart or lung disease; for the newborn altered cerebrovascular blood flow with intracerebral hemorrhage and pulmonary hypertension. 2) They also will contribute to understanding mechanisms of cardiovascular disorders and renatal "programming" of adult disease. 3) Finally, they are relevant to understanding mechanisms of diseases such as: Acute Mountain Sickness, Preeclampsia, and High Altitude Cerebral and Pulmonary Edema.

本项目的总体主题是了解高海拔长期缺氧（LTH）的适应改变胎儿和成人血管系统的基本机制。我们还将研究这些机制与从胎儿到成人的发展。这个项目是广泛的基础上，多学科，并垂直整合使用生理，细胞，生物化学和分子的方法。基于二十年的研究，我们将在适应高海拔的绵羊中测试一些假设。总体假设是，高海拔、长期缺氧与继发于改变的α-肾上腺素能受体（或AR）亚型和/或特异性蛋白激酶C亚型（PKC）介导的下游Ca 2+依赖性和Ca非依赖性信号转导途径的脑血管收缩反应的变化相关。一个相关的假设是，LTH显着改变Oi-AR亚型和特定的PKC同工酶介导的原癌基因和基因的表达，代表血管平滑肌的“合成”和/或“增殖”表型，与成人的“收缩”表型。四个具体的目标将检查LTH在脑动脉或 肾上腺素能介导的薄和厚肌丝的信号转导，例如，1)O 1-AR、PKC亚型、细胞外信号调节激酶（ERK）和下游效应蛋白的作用，2）质膜和肌浆网Ca 2+通道的作用，3）血管表型的基因调节，和4）信号传导途径基因/蛋白发现。对于这些研究，我们将利用激动剂诱导的收缩性和细胞内[Ca 2 +]，Western免疫生物素，RT-PCR，RNAi沉默，共聚焦显微镜，2D-凝胶-质谱和基因微阵列/途径分析。科学地说，这些研究将增加我们对胎儿和成人脑血管适应LTH的基本机制的理解。他们还将说明从胎儿到成人的发展方面。在临床上，这些研究涉及至少三个关键问题。1)对于胎儿和新生儿，它们与长时间缺氧的反应有关，如生活在高海拔地区的妇女，以及吸烟或贫血的妇女，患有心脏或肺部疾病的妇女;对于新生儿，脑血管血流改变，伴有脑出血和肺动脉高压。2)他们还将有助于了解心血管疾病和成人疾病的先天“编程”机制。3)最后，它们与理解疾病的机制有关，例如：急性高原病，先兆子痫和高海拔脑水肿和肺水肿。