Alcohol and Maternal Uterine Vascular Adaptations in Pregnancy

描述(申请人提供)：怀孕期间的酒精和母体子宫血管适应性怀孕期间滥用酒精会导致胎儿酒精谱障碍(FASD)，这是一种终生残疾，其特征是一系列生长和发育缺陷。目前估计FASD在美国学龄儿童中的患病率约为2%-5%。努力成功地预防或改善乙醇的致畸作用 对酒精涉及多个器官系统的复杂作用机制的有限了解，至少在一定程度上阻碍了这一进程。正常妊娠与主要的子宫循环适应有关，而子宫循环适应直接与胎儿生长、新生儿出生体重有关 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 酒精研究的重点是大脑/行为缺陷，而对关键的妊娠子宫血管适应问题关注较少。在此，我们展示了新的初步数据，酒精损害了酒精模型大鼠妊娠子宫循环适应的精细调节。在此，我们假设怀孕期间长期酗酒通过内皮一氧化氮(NO)系统失调来损害母体子宫动脉的血管适应性。目的#1将检验孕期酗酒暴露导致内皮依赖性母体子宫动脉松弛受损的假说。遵循Binger范式，我们将评估生长指数，并利用线状肌图术研究生理盐水对照组、成对喂养的营养对照组和酒精大鼠的内皮完整/裸露血管中激动剂诱导的子宫动脉松弛。目的#2研究酗酒是否通过与前列环素/内皮源性超极化因子相关的内皮源性NO损伤子宫动脉的舒张性，减少子宫动脉NO的产生，内皮型一氧化氮合酶(ENOS)的表达，并损害eNOS的多位点磷酸化。目的#3研究酗酒是否通过ERK/AMPK途径降低子宫动脉NO，降低兴奋性Pser1177eNOS水平，增加抑制性Pth495eNOS水平，以及酒精对内皮细胞[Ca+2]i瞬变的影响。在目标2和目标3中，我们将评估阻断主要血管扩张途径后的子宫动脉松弛，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/多部位磷酸化调节途径拮抗剂的机制研究。利用高速激发/发射波长切换系统同时进行[Ca~(2+)]I-NO荧光成像。我们的建议探索了妊娠酒精研究的新前沿，开发了第一个酗酒诱导子宫动脉适应的机制框架，并在体内模型中确定了酒精靶标。与NIAAA 2014财年战略计划相一致，我们的建议利用了强大的方法，并向Fas领域提出了一个新的母体包容范式，并预测更有效的干预将需要针对母体系统，特别是关键的子宫循环的创新药理学靶向，以便预测和提出一种真正有希望作为预防策略的治疗方法。

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.