Pregnancy/NO Induced Changes in UAE Ca2+Signaling

怀孕/NO 引起的阿联酋 Ca2 信号变化

• 批准号: 8463585
• 负责人: IAN M. BIRD
• 金额: $ 34.97万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8463585
• 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

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. PUBLIC HEALTH RELEVANCE: Preeclampsia (PE) is a potentially devastating disease of pregnancy that can threaten the life of both mother and the unborn child. It affects as many as 20% of the population. Even mild PE is dangerous and can cause preterm (early) birth and this then gives the baby an increased risk to develop several diseases such as blood pressure, diabetes and obesity as an adult. It is believed that in PE the cells lining the uterine artery blood vessels fail to increase their function as would otherwise happen in normal pregnancy. We have evidence to suggest normal adaptation is due to electrical changes and their failure may be 'fixed' with a specific drug NS309. The aim of this project is to find out if our suspicions are correct by testing in an animal model. If we are right we may be able to come up with a treatment plan fro PE pregnancy. Having more complete knowledge of how cell function is regulated in pregnancy will certainly take us one step closer to overcoming this potentially devastating disease that effects so many in this country and beyond.

描述（由申请人提供）：在美国，先兆子痫（PE）对多达20％的怀孕产生了不利影响。最早的事件之一是母体血管内皮血管舒张功能的失败以适应，正常妊娠相关的增加，无钝或丢失。在以前的资金期间，我们使用了[Ca2+] I的双重成像，而在新鲜孤立的子宫动脉内皮中则使用了NO，以表明在正常怀孕中，CA2+信号的增强是提高NO产生的主要原因，并且这种增强取决于特定的变化。 CA2+信号传导介质的功能。通过TRPC通道发生持续的Ca2+进入UA内皮，但是我们的初步数据还表明，TRPC通道的妊娠增强，依次取决于CX43间隙连接通信，并且细胞的这种耦合可能会在电气水平上发生。我们的数据进一步表明，KCA类型通道的膜超极化连接增加的间隙连接功能，以增强怀孕的TRPC功能。与此相一致，UAEC对KCA功能的拮抗剂（Apamin和TRAM34）表现出敏感性，导致Ca2+信号传导的降低。如果该模型是正确的，那么可以通过使用SK或IK型KCA通道激动剂来实现PE妊娠中Ca2+信号传导失败的未来治疗。在这种续签中，我们建议测试该模型，从培养物中的孤立细胞转变为整个血管，从体内到整个动物研究。我们将检查1）在低密度与高密度的NP和P-UAEC中响应ATP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的效果-34或Ca2+突发活动上的选择性激动剂NS309在此响应中暗示KCA通道； 2）检查这些相同的拮抗剂和激动剂对Ca2+的变化，膜电位变化的影响，并且在完整的UA容器中没有生产； 3）确定KCA激动剂NS309是否与ATP/UTP在怀孕期间体内子宫动脉的血流无依赖性增加以及SK和/或IK类型的KCA通道中介导此介导的程度的程度相当影响。通过这种方式，我们将最新的关于血液妊娠适应的机理知识的进步现在确定了使用KCA通道激动剂在妊娠中最大化这种反应，尤其是PE复杂的有效性。成功可能会对改善这种正常适应性反应失败并且对当前疗法具有抵抗力的正常适应性反应的治疗的策略产生重大影响。 公共卫生相关性：先兆子痫（PE）是一种可能造成毁灭性的怀孕疾病，可能威胁母亲和未出生的孩子的生命。它影响了多达20％的人口。即使是轻度的PE也很危险，可能会导致早产（早产），然后这使婴儿增加了多种疾病的风险，例如血压，糖尿病和成年人的肥胖症。据信，在PE中，衬有子宫动脉血管的细胞无法提高其功能，否则在正常怀孕中会发生。我们有证据表明正常适应性是由于电动变化造成的，并且可能使用特定的药物NS309“固定”其故障。该项目的目的是确定我们的怀疑是否正确通过在动物模型中进行测试。如果我们是对的，我们可能会提出一个从妊娠中的治疗计划。对怀孕中细胞功能的调节方式有了更完整的了解，肯定会使我们更近一点克服了这种潜在的毁灭性疾病，在这个国家及以后会影响很多。