Endothelium-Derived Vasodilators in Pregnancy

妊娠期内皮衍生血管扩张剂

• 批准号: 7419016
• 负责人: RONALD R MAGNESS
• 金额: $ 33.08万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-07-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7419016
• 关键词: 1-Phosphatidylinositol 3-Kinase; Address; Agonist; Arteries; Birth Weight; Blood flow; Cell Communication; Cell Line; Cell Separation; Cell model; Cells; Coupling; Cultured Cells; Data; Disease; Elevation; Employee Strikes; Endothelial Cells; Endothelium; Estrogen Receptors; Estrogens; Fetal Growth; Fetal Growth Retardation; Functional disorder; Gene Expression; Genomics; Image; In Vitro; Lonomycin; Luteal Phase; MAP Kinase Gene; MAPK Signaling Pathway Pathway; Measures; Mediating; Microscope; Mitogen Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Modeling; Molecular; Nitric Oxide; Ovarian; Pathway interactions; Patient currently pregnant; Personal Satisfaction; Phosphoinositide-3-Kinase, Catalytic, Gamma Polypeptide; Phosphorylation Inhibition; Phosphotransferases; Physiological; Play; Pre-Eclampsia; Pregnancy; Principal Investigator; Production; Proto-Oncogene Proteins c-akt; Recruitment Activity; Role; Sheep; Signal Pathway; Signal Transduction; Slide; Third Pregnancy Trimester; Time; Vasodilation; Vasodilation disorder; Vasodilator Agents; Woman; base; cell growth regulation; fetal; in vitro Model; in vivo; non-genomic; novel; physiologic model; programs; proliferative phase Menstrual cycle; response; shear stress

DESCRIPTION (provided by applicant): Interactions between estrogen and NO to regulate UBF at the maternal-fetal placental interface are critically important to fetal growth. Pregnant (P) and Follicular (Fol) Nonpregnant (NP) Uterine Artery Endothelial Cells (UAEC) undergo adaptation compared to Luteal (Lut) NP-UAEC; i.e. responses seen in freshly isolated UA Endothelium maintaining upon numerous passages. These include ERK 1/2 -MAPK coupling to eNOS activation for substantial ATP-stimulated NO production (P > Fol-NP> Lut-NP) and the role estrogen may have to alter Ca2+ mediated NO. Shear stress and/or estrogen are both very potent stimulators of NO synthesis and/or eNOS expression and together they elicit synergistic activation of the ERK 1/2 MAPK signaling pathway in P-UAEC. Elevations in blood flow also substantially increase the laminar shear stress. Other signaling pathways (e.g. PI3 Kinase) may play an additional convergent role in the in vivo situation. Overall hypothesis: Physiologic states of High DBF/shear stress and elevated estrogen (e.g. P and Fol) causes "Programmed Adaptation" of the UAEC to modulate important endothelial functions associated with NO production. Specific emphasis is given to mechanisms that increase eNOS activation, eNOS expression, UAEC signaling kinases (e.g. ERK 1/2 and/or. PIS Kinase), and cell-cell communication (measured as synchronized Ca2+ bursts). Aim I- Physiologic Models: Using In V/Vo-derived freshly isolate UA Endothelium (Passage 0) from Pregnant, Cycling (Fol and Lut) and Ovariectomized (Vehicle vs Estrogen-treated) sheep, studies will be performed evaluating cellular regulation (nongenomic and genomic) by which Estrogen alters basal, ATP and lonomycin eNOS activation by directly imaging intracellular NO production simultaneously with [Ca2+]i/Ca2+ bursts and evaluating the role of ERs and converging signaling intermediate (ERK1/2 MAPK and/or PIS Kinase) pathways. Aim II- Shear Stress Models: Effects of prolonged Laminar Shear Stress [0 (static), 3 & 15 dynes/cm2] with and without E2(3 on "Programmed UAEC Adaptation" of long term cultured (Passage 4-5) Lut NP-UAEC, Fol NP- UAEC, and P-UAEC. The specific markers of adaptation that are "programmed" from the time isolation we focus on include: a) eNOS expression and ATP/lonomycin-stimulated eNOS activation and [Ca2+]i/Ca2+ bursts and b) ATP/lonomycin-stimulated ERK1/2 and AKT phosphorylation; and inhibition of ERK-MAPK and/or PIS Kinase mediated stimulated NO production and [Ca2+]i/Ca2+ bursts. We hypothesize that inclusion of both shear stress and estrogen will be necessary to restore the specific markers and response magnitude "programmed" into UAECs by high flow and estrogen during Pregnancy and in the Follicular phase defined in Aim I. Data thus derived will provide the first mechanistic framework for understanding interactions between shear stress and estrogen to regulate UAEC NO and converging signaling mechanisms this. Our recent preliminary data demonstrate that uterine artery shear stress is indeed reduced in pregnancies complicated by preeclampsia. Therefore these mechanisms are likely also to be dysfunctional during preeclampsia with or without IUGR.

描述（由申请人提供）：雌激素和NO之间的相互作用调节母胎胎盘界面的UBF对胎儿生长至关重要。与黄体（Lut） NP-UAEC相比，妊娠(P)和卵泡（Fol）非妊娠（NP）子宫动脉内皮细胞（UAEC）经历了适应性；即在新鲜分离的UA中观察到的反应，内皮维持在许多传代上。这些包括ERK 1/2 -MAPK偶联到eNOS激活，以产生大量atp刺激的NO (P > foll - np > Lut-NP)，以及雌激素可能必须改变Ca2+介导的NO的作用。剪切应力和/或雌激素都是NO合成和/或eNOS表达的有效刺激物，它们共同引发P-UAEC中ERK 1/2 MAPK信号通路的协同激活。血流量的增加也大大增加了层流剪切应力。其他信号通路（如PI3激酶）可能在体内情况下发挥额外的收敛作用。总体假设：高DBF/剪切应力和雌激素（如P和Fol）升高的生理状态导致UAEC的“程序性适应”，以调节与NO产生相关的重要内皮功能。特别强调了增加eNOS激活、eNOS表达、UAEC信号激酶（如erk1 /2和/或）的机制。PIS激酶)和细胞-细胞通讯（测量同步Ca2+爆发）。目标1 -生理模型：使用V/ vo衍生的新鲜分离的UA内皮（传代0），这些内皮来自怀孕、循环（Fol和Lut）和卵巢切除（对照雌激素处理）的绵羊，研究将评估雌激素通过细胞调节（非基因组和基因组）改变基础、通过直接成像细胞内NO生成与[Ca2+]i/Ca2+爆发同时发生，并评估er和聚合信号中间体（ERK1/2 MAPK和/或PIS激酶）途径的作用来激活ATP和洛霉素eNOS。Aim II-剪切应力模型：有和没有E2(3)的长时间层状剪切应力[0（静态），3和15 dynes/cm2]对长期培养的Lut NP-UAEC， Fol NP-UAEC和P-UAEC“程序化适应”的影响。我们关注的时间隔离中“程序化”的特定适应标记包括：a) eNOS表达和ATP/lonomycin刺激的eNOS激活和[Ca2+]i/Ca2+爆发；b) ATP/lonomycin刺激的ERK1/2和AKT磷酸化；抑制ERK-MAPK和/或PIS激酶介导的刺激NO产生和[Ca2+]i/Ca2+爆发。我们假设，剪切应力和雌激素的结合对于恢复妊娠期间和卵泡期高流量和雌激素“编程”到UAEC中的特定标记物和反应强度是必要的。由此得出的数据将为理解剪切应力和雌激素之间的相互作用提供第一个机制框架，以调节UAEC NO及其趋同信号机制。我们最近的初步数据表明，子宫动脉剪应力确实减少妊娠合并先兆子痫。因此，这些机制也可能在有或没有IUGR的子痫前期功能失调。