Pregnancy/NO Induced Changes in UAE Ca2+Signaling

怀孕/NO 引起的阿联酋 Ca2 信号变化

基本信息

批准号：
8707534
负责人：
IAN M. BIRD
金额：
$ 36万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-05-01 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8707534
关键词：
ATP Receptors Abbreviations Adult Adverse effects Affect Agonist Animal Model Animals Apamin Arteries Birth Blood Pressure Blood Vessels Blood flow Cell Count Cell Line Cell membrane Cell physiology Cells Communication Connexin 43 Connexins Country Coupled Coupling Data Dependence Diabetes Mellitus Disease Electrophysiology (science)Endothelial Cells Endothelium Event Failure Fetal Growth Retardation Funding Future GTP-Binding Proteins Gap Junctions Growth Factor Human Image In Vitro Individual Inositol Ion Channel Ions Knowledge Life Measures Mediating Mediator of activation protein Membrane Membrane Potentials Modeling Mothers Movement Nitric Oxide Obesity One-Step dentin bonding system P2X-receptor Perfusion Pharmaceutical Preparations Phospholipase C Phosphorylation Placenta Play Population Potassium Channel Pre-Eclampsia Pregnancy Probability Process Production Relative (related person)Resistance Risk Role Series Signal Transduction Smooth Muscle Myocytes TRPC3 ion channel Testing VEGF165 Vascular Endothelial Growth Factors Vascular Endothelium Vascular Smooth Muscle Vasodilation Woman Work density functional restoration human NOS3 protein improved in vivo inhibitor/antagonist myometrium pregnant pressure public health relevance receptor response success therapeutic target treatment planning unborn child voltage

项目摘要

DESCRIPTION (provided by applicant): Preeclampsia (PE) adversely affects as many as 20% of pregnancies in the USA. One of the earliest events is the failure of maternal vascular endothelium vasodilatory function to adapt, with the normal pregnancy-associated increase in NO being blunted or lost. In our previous funding period we used dual imaging of [Ca2+]i and NO in freshly isolated uterine artery endothelium to show that in normal pregnancy, enhancement of Ca2+ signaling is the primary cause of enhanced NO production, and such enhancement depends on specific changes in function of Ca2+ signaling mediators. Sustained Ca2+ entry into UA endothelium occurs through TRPC channels but our preliminary data also suggests the pregnancy enhanced opening of TRPC channels in turn depends on CX43 Gap junction communication, and that such coupling of cells may occur at an electrical level. Our data further suggests membrane hyperpolarization by KCa type channels connects increased Gap junction function to enhance TRPC function in pregnancy. Consistent with this, UAEC show sensitivity to antagonists of KCa function (Apamin and Tram34) with a resulting reduction in Ca2+ signaling. If this model is correct then future therapy for failed adaptation of Ca2+ signaling in PE pregnancy could be achieved by the use of SK or IK type KCa channel agonists. In this renewal we propose to test this model, moving from isolated cells in culture to whole vessels ex vivo to whole animal studies. We will examine 1) Changes in membrane potential and KCa current activity in response to ATP in NP- and P-UAEC in cells at low vs high density, dependence of these changes on gap junction function, and determine the effect of antagonists Apamin and Tram-34 or the selective agonist NS309 on Ca2+ burst activity to implicate KCa channels in this response; 2) Examine the effects of these same antagonists and agonists on changes in Ca2+, membrane potential and NO production in intact UA vessels ex vivo; 3) determine if the KCa agonist NS309 is comparable to the effects of ATP/UTP in mediating NO dependent increases in blood flow in uterine artery in vivo during pregnancy, and also the extent to which SK and/or IK types of KCa channel mediate this effect. In this way we take our recent advances in mechanistic knowledge of pregnancy adaptation of blood flow to now establish the validity of using KCa channel agonists to maximize this response in vivo in pregnancy and particularly those complicated by PE. Success could have major impact on strategies to improve treatment of human PE pregnancy where this otherwise normal adaptive response has failed and is resistant to current therapies.

描述（由申请人提供）：在美国，先兆子痫 (PE) 对多达 20% 的妊娠产生不利影响。最早的事件之一是母体血管内皮血管舒张功能无法适应，正常妊娠相关的一氧化氮（NO）增加被减弱或丧失。在我们之前的资助期间，我们使用了新鲜分离的子宫动脉内皮中的[Ca2+]i和NO双重成像来表明，在正常妊娠中，Ca2+信号传导的增强是NO产生增强的主要原因，并且这种增强取决于Ca2+信号传导介质的功能。 Ca2+ 持续进入 UA 内皮是通过 TRPC 通道发生的，但我们的初步数据还表明妊娠增强的 TRPC 通道开放反过来又取决于 CX43 间隙连接通信，并且这种细胞耦合可能发生在电水平上。我们的数据进一步表明，KCa 型通道的膜超极化与间隙连接功能的增强有关，从而增强妊娠期间的 TRPC 功能。与此一致的是，UAEC 对 KCa 功能拮抗剂（Apamin 和 Tram34）表现出敏感性，从而导致 Ca2+ 信号传导减少。如果这个模型是正确的，那么未来对 PE 妊娠中 Ca2+ 信号适应失败的治疗可以通过使用 SK 或 IK 型 KCa 通道激动剂来实现。在这次更新中，我们建议测试这个模型，从培养中的分离细胞转移到离体的整个血管到整个动物研究。我们将检查 1) 低密度与高密度细胞中 NP- 和 P-UAEC 中 ATP 响应的膜电位和 KCa 电流活性的变化，这些变化对间隙连接功能的依赖性，并确定拮抗剂 Apamin 和 Tram 的作用-34 或选择性激动剂 NS309 对 Ca2+ 爆发活性的影响，表明 KCa 通道参与了该反应； 2) 离体检查这些拮抗剂和激动剂对完整 UA 血管中 Ca2+、膜电位和 NO 产生变化的影响； 3)确定KCa激动剂NS309在介导妊娠期间体内子宫动脉血流量NO依赖性增加方面的作用是否与ATP/UTP相当，以及KCa通道的SK和/或IK类型介导这种作用的程度影响。通过这种方式，我们利用妊娠期血流适应机制方面的最新进展，确定了使用 KCa 通道激动剂在妊娠期（尤其是那些并发 PE 的妊娠期）体内最大化这种反应的有效性。成功可能会对改善人类早泄妊娠治疗的策略产生重大影响，因为这种正常的适应性反应已经失败，并且对当前的治疗有抵抗力。