Endothelium-Derived Vasodilators in Pregnancy

妊娠期内皮衍生血管扩张剂

• 批准号: 7871380
• 负责人: RONALD R MAGNESS
• 金额: $ 35.07万
• 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/7871380
• 关键词: 1-Phosphatidylinositol 3-Kinase; Address; Agonist; Arteries; Birth Weight; Blood flow; Cell Communication; Cell Culture Techniques; Cell Line; Cell Separation; Cell model; Cells; Coupling; Data; Disease; 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; 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; Vasodilator Agents; Woman; base; cell growth regulation; fetal; in vitro Model; in vivo; non-genomic; novel; physiologic model; pregnant; programs; proliferative phase Menstrual cycle; response; response marker; 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(申请人提供)：雌激素和一氧化氮之间的相互作用，以调节母胎胎盘界面的UBF，对胎儿生长至关重要。妊娠(P)和卵泡(Fol)非妊娠(NP)子宫动脉内皮细胞(UAEC)与黄体(Lut)NP-UAEC相比经历适应；即在新鲜分离的UA内皮细胞维持多代反应。其中包括ERK 1/2-MAPK与eNOS的偶联，以产生大量的ATP刺激的NO(P&gT；Fol-NP&GT；Lut-NP)，以及雌激素可能在改变钙离子介导的NO中所起的作用。切应力和/或雌激素都是非常有效的NO合成和/或eNOS表达的刺激物，它们共同诱导P-UAEC中ERK 1/2 MAPK信号通路的协同激活。血流的升高也大大增加了层流切应力。其他信号通路(如PI3Kinase)可能在体内情况下发挥额外的汇聚作用。总体假设：高DBF/切应力和雌激素升高(如P和Fol)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信号通路(如ERK 1/2和/或)的机制。PIS Kinase)和细胞-细胞通讯(以同步的钙离子爆发来测量)。目的I-生理学模型：在V/Vo来源的新鲜分离的妊娠、周期(Fol和Lut)和去卵巢(载体与雌激素处理)绵羊的UA内皮(0代)中，通过直接成像细胞内NO的产生与[Ca+]i/Ca~(2+)的同时，评价ERs和汇聚的信号中间体(ERK1/2 MAPK和/或PiS Kinase)通路的作用，以评估雌激素改变基础、ATP和lonomycin eNOS激活的细胞调控(非基因组和基因组调控)。目的II-剪应力模型：长期层流剪应力[0(静态)，3~15dynes/cm2]加和不加E2(3)对长期培养(第4-5代)Lut NP-UAEC、Fol NP-UAEC和P-UAEC的“程序性UAEC适应”的影响。从我们所关注的时间分离中“程序化”的适应的特异性标志物包括：a)eNOS的表达和ATP/lonomycin刺激的eNOS的激活和[Ca+]i/Ca~(2+)爆发；b)ATP/lonomycin刺激的ERK1/2和AKT的磷酸化；以及抑制ERK-MAPK和/或PiS Kinase介导的NO的产生和[Ca~(2+)]i/Ca~(2+)爆发。我们假设，包括切应力和雌激素将是必要的，以恢复特定的标志物和反应幅度“编程”到UAECs在妊娠和卵泡期定义的目标I。数据将提供第一个机制框架，以了解切应力和雌激素之间的相互作用，以调节UAEC NO和汇聚的信号机制。我们最近的初步数据表明，妊娠合并子痫前期患者子宫动脉切应力确实降低。因此，在有或没有IUGR的先兆子痫期间，这些机制也可能功能失调。