Chronic hypoxia, AMPK activation and uterine artery blood flow

慢性缺氧、AMPK 激活与子宫动脉血流

• 批准号: 9925655
• 负责人: Colleen Glyde Julian
• 金额: $ 32.27万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9925655
• 关键词: Adenosine Monophosphate; Affect; Agonist; Altitude; Arteries; Biometry; Biopsy; Blood Vessels; Blood flow; Blood specimen; Caliber; Catalytic Domain; Cesarean section; Chronic; DNA Methylation; Data; Discipline of obstetrics; Endothelium; Enzymes; Epigenetic Process; Fetal Growth; Fetal Growth Retardation; Fetal Tissues; Frequencies; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Growth; Homeostasis; House mice; Human; Hypoxia; Infant; Intervention; Ischemia; Link; Measures; Mediating; Metabolic; Metformin; Methylation; Mus; Myometrial; NIH Program Announcements; Peripheral Blood Mononuclear Cell; Pharmacology; Phosphotransferases; Placenta; Plasma; Play; Positioning Attribute; Postpartum Period; Pre-Eclampsia; Pregnancy; Process; Prospective Studies; Protein Isoforms; Protein Kinase; Proteins; Protocols documentation; Public Health; Publications; Regulation; Risk; Role; Sea; Signal Pathway; Signal Transduction; Signaling Protein; Single Nucleotide Polymorphism; Testing; Thoracic aorta; Time; Tissues; Uteroplacental Circulation; Uterus; Vascular Smooth Muscle; Vasodilation; Vasodilator Agents; Weight; Western Blotting; Woman; Work; cohort; drug development; epigenetic regulation; fetal; functional outcomes; health of the mother; human data; improved; in utero; inhibitor/antagonist; innovation; mRNA Expression; mouse model; novel; pregnancy disorder; pregnant; prevent; recruit; residence; response; upstream kinase

Project Summary High altitude residence (HA, >2500 m) increases the frequency of preeclampsia (PreE) and intrauterine growth restriction (IUGR) 3-fold. Since hypoxia is a common cause, HA studies are uniquely positioned to evaluate the mechanistic role of hypoxia, discover new treatments, and alleviate the “almost complete lack of drug development for obstetric indications” noted in PAR-13-389. Adenosine monophosphate kinase (AMPK) is a regulator of metabolic homeostasis that also affects vascular growth and function under hypoxia. We have shown that AMPK plays important roles in the regulation of human uterine artery (UtA) blood flow and fetal growth at HA, and that AMPK activation has vasodilator effects in isolated murine UtA that are potentiated by hypoxia. Our central hypothesis is that AMPK activation promotes vasodilation in the uteroplacental circulation in response to hypoxia to raise UtA blood flow and improve fetal growth at HA. Because hypoxia leads to epigenetic changes that alter the expression of genes regulating of AMPK activity and fetal growth, we propose that DNA methylation influences these AMPK-mediated processes. We present new murine and human data to show that hypoxia a) raises placental phosphorylated (P)- relative to total AMPK levels and the P- as well as the total protein levels of AMPK targets in thoracic aorta and placenta; b) increases the expression of key enzymes activating AMPK in UtA and placenta; and c) alters methylation-expression relationships of AMPK- signaling genes important for fetal growth. We will test our central hypothesis by determining: 1. In Aim 1, the effect of HA pregnancy on AMPK signaling and its relationship to UtA blood flow and fetal growth in humans. We will recruit 102 healthy residents of low altitude (LA, 1600 m, n=53) or HA (3000 m, n=49); measure UtA blood flow and fetal biometry longitudinally; and determine the activation of AMPK, its upstream regulators and downstream targets, the expression levels and DNA methylation of relevant genes in peripheral blood mononuclear cells (PBMCs), and plasma levels of AMPK regulators. 2. In Aim 2, the effects of hypoxia on AMPK activation in human myometrial artery (MA) and placenta, and on MA vasoreactivity. In women participating in Aim 1 who deliver by elective C-section (n=19/altitude), we will obtain myometrial biopsies and placentas to determine a) the activation, expression and DNA methylation status of AMPK, its well-established regulators and downstream targets; b) the effect of HA pregnancy on MA vasoreactivity; and c) whether hypoxia potentiates vasodilator effects of AMPK activation in MA. 3. In Aim 3, the role of AMPK in regulating uteroplacental blood flow and fetal growth in response to hypoxia in mice. Pregnant mice housed at sea level (SL) or HA will be treated with the AMPK activator (AICAR), inhibitor (Compound C), or vehicle (control) for determining the separate and combined effects of hypoxia and AMPK activation on AMPK signaling in UtA, placental and fetal tissues; UtA vascular reactivity and blood flow; and fetal growth.

项目摘要 高海拔居住（HA，> 2500 m）增加了子痫前期（PREE）和intratorine生长的频率 限制（IUGR）3倍。由于缺氧是常见原因，因此HA研究是独特的，以评估 缺氧的机械作用，发现新疗法并减轻“几乎完全缺乏药物 产科适应症的开发”在Par-13-389中指出。腺苷一磷酸激酶（AMPK）是一个 我们有代谢稳态的调节剂，它也影响缺氧下的血管生长和功能。 表明AMPK在人子宫动脉（UTA）血流和胎儿的调节中起重要作用 HA的增长，AMPK激活在孤立的鼠UTA中具有血管扩张剂的作用，这些作用是由 缺氧。我们的中心假设是AMPK激活促进子宫循环中的血管舒张 响应缺氧以增加UTA血流并改善HA时胎儿生长。因为缺氧导致 表观遗传变化改变了调节AMPK活性和胎儿生长的基因表达，我们提出 DNA甲基化会影响这些AMPK介导的过程。我们向 证明缺氧a）相对于总AMPK水平以及P-以及P-以及 胸主动脉和plapeta中AMPK靶标的总蛋白质水平； b）增加钥匙的表达 在UTA和PLACETA中激活AMPK的酶； c）改变AMPK-的甲基化表达关系 信号基因对胎儿生长很重要。我们将通过确定： 1。在AIM 1中，HA妊娠对AMPK信号传导及其与UTA血流和胎儿的关系 人类的增长。我们将招募102名低海拔（LA，1600 m，n = 53）或HA（3000 m，3000 m， n = 49）;纵向测量UTA血流和胎儿生物特征；并确定AMPK的激活 上游调节剂和下游靶标，相关的表达水平和DNA甲基化 外周血单核细胞中的基因（PBMC）和AMPK调节剂的血浆水平。 2。在AIM 2中，缺氧对人肌层动脉（MA）和PLACETA的AMPK激活以及对AMPK激活的影响，以及对 MA血管反应性。在参加AIM 1的女性中，她通过选修剖腹产（n = 19/高度） 获得肌分活检和片剂，以确定a）激活，表达和DNA甲基化 AMPK的地位，其成熟的监管机构和下游目标； b）HA怀孕对 马血管反应性； c）缺氧是否会增强MA中AMPK激活的血管扩张效应。 3。在AIM 3中，AMPK在控制子宫流体流量和胎儿生长中的作用对缺氧 在老鼠中。在海平面（SL）或HA的怀孕小鼠将用AMPK激活剂（AICAR）治疗， 抑制剂（Compound C）或用于确定缺氧的单独和组合作用的媒介物（对照） 在UTA，斑点和胎儿组织中对AMPK信号传导的AMPK激活； UTA血管反应性和 血流（量;和胎儿生长。