Alcohol and Maternal Uterine Vascular Adaptations in Pregnancy

描述（由申请人提供）：妊娠期酒精和母体子宫血管适应性母亲在怀孕期间滥用酒精可导致胎儿酒精谱系障碍（FASD），这是一种终身残疾，其特征是一系列生长和发育缺陷。目前估计FASD患病率约为2-5%的年轻学生在美国。成功预防或改善乙醇致畸作用的努力 由于对酒精涉及多个器官系统的复杂作用机制的了解有限，至少部分地受到阻碍。正常妊娠与子宫循环系统的主要适应性相关，这些适应性直接关系到胎儿生长、新生儿出生体重 和生存胎儿酒精综合征的一个主要特征是生长受限，但传统上 酒精研究集中在大脑/行为缺陷上，而很少关注关键的妊娠子宫血管适应。在此，我们展示了新的初步数据，即酒精损害大鼠酒精模型妊娠子宫循环适应的精细调节。我们在此假设，慢性酗酒暴露在怀孕期间损害母体子宫动脉血管适应通过内皮一氧化氮（NO）系统失调。目标1将检验妊娠期酗酒暴露导致内皮依赖性母体子宫动脉舒张受损的假设。根据狂欢模式，我们将评估生长指数，并利用钢丝肌造影术研究激动剂诱导的子宫动脉舒张的内皮完整/裸露的血管生理盐水对照，成对喂养的营养控制，和酒精大鼠。目标#2将测试酗酒是否通过相对于前列环素/内皮衍生超极化因子的内皮衍生NO损害子宫动脉舒张，降低子宫动脉NO产生、内皮NO合酶（eNOS）表达，并损害eNOS多位点磷酸化。目的#3将测试酗酒是否通过ERK/AMPK途径降低子宫动脉NO，降低兴奋性Pser 1177 eNOS水平，并增加抑制性Pthr 495 eNOS水平，以及酒精对内皮[Ca+2]i瞬变的影响。在目标#2和#3中，我们将评估阻断主要血管舒张通路后的子宫动脉舒张，并通过RT-PCR、免疫印迹、组织学、荧光成像和分光光度法进行有/无eNOS活性/多位点磷酸化调节通路拮抗剂的机制研究。同时[Ca 2 +]i-NO荧光成像将利用高速激发/发射波长切换系统进行。我们的提案通过开发第一个酗酒诱导的子宫动脉适应机制框架和在体内模型中识别酒精靶点，探索了妊娠期酒精研究的新前沿。与NIAAA FY 14战略计划一致，我们的提案利用了强大的方法，并为FAS领域提供了一种新的母亲包容性范例，并预测更有效的干预将需要创新的药物靶向母体系统，特别是关键的子宫循环，以预测和提出一种具有真实的预防策略的治疗。

DESCRIPTION (provided by applicant): Alcohol & Maternal Uterine Vascular Adaptations in Pregnancy Maternal alcohol abuse during pregnancy can result in Fetal Alcohol Spectrum Disorders (FASD), a lifelong disability characterized by a range of growth and developmental deficits. Current estimates of FASD prevalence is about 2-5% among young school children in the United States. Efforts to successfully prevent or ameliorate the teratogenic effects of ethanol have been impeded, at least in part, by a limited understanding of alcohol's complex mechanisms of action involving multiple organ systems. A normal pregnancy is associated with major uterine circulatory adaptations that directly relates to fetal growth, neonatal birth weights and survival. A cardinal feature of fetal alcohol syndrome is growth restriction, but traditionally alcohol studies have focused on brain/behavioral deficits, and little attention has been paid to critical gestational uterine vascular adaptations. We herein show novel preliminary data that alcohol impairs the exquisite regulation of gestational uterine circulatory adaptations in rat bing alcohol model. We herein hypothesize that chronic binge alcohol exposure during pregnancy impairs maternal uterine artery vascular adaptations via endothelial nitric oxide (NO) system dysregulation. Aim#1 will test the hypothesis that binge alcohol exposure in pregnancy leads to impaired endothelium-dependent maternal uterine artery relaxation. Following binge paradigm, we will assess growth indices, and utilize wire myography to study agonist-induced uterine artery relaxation in endothelium-intact/denuded vessels from saline control, pair-fed nutrition control, and alcohol rats. Aim#2 will test if binge alcohol impairs uterine artery relaxation via endothelium-derived NO relative to prostacyclin/endothelium-derived hyperpolarizing factor, decreases uterine artery NO production, endothelial NO synthase (eNOS) expression, and impairs eNOS multi-site phosphorylation. Aim#3 will test if binge alcohol decreases uterine artery NO, decreases excitatory Pser1177eNOS levels, and increases inhibitory Pthr495eNOS levels via ERK/AMPK pathway, and alcohol effects on endothelial [Ca+2]i transients. In Aims#2 and #3, we will assess uterine artery relaxation after blocking major vasodilatory pathways and conduct mechanistic studies with/without eNOS activity/multi-site phosphorylation-regulating pathway antagonists via RT-PCR, immunoblotting, histology, fluorescent imaging and spectrophotometry. Simultaneous [Ca2+]i-NO fluorescent imaging will be performed utilizing high-speed excitation/emission wavelength switching system. Our proposal explores a new frontier of gestational alcohol research by developing the first mechanistic framework for binge alcohol-induced uterine artery adaptations and identifying alcohol targets in an in vivo model. In alignment with NIAAA FY14 Strategic Plan, our proposal utilizes powerful methods and presents a new maternal-inclusive paradigm to the FAS field and predicts that a more effective intervention will require innovative pharmacologic targeting of maternal systems, especially the critical uterine circulation, in order to predict and propose a therapy that will have a real promie as a preventive strategy.