Endothelium-Derived Vasodilators in Pregnancy

妊娠期内皮衍生血管扩张剂

• 批准号: 7880324
• 负责人: RONALD R MAGNESS
• 金额: $ 1.83万
• 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/7880324
• 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

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 (Foland 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(3on "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 demonstrates 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.

雌激素和 NO 之间的相互作用对于调节母胎胎盘界面的 UBF 至关重要 对胎儿生长很重要。妊娠 (P) 和滤泡 (Fol) 非妊娠 (NP) 子宫动脉内皮细胞 (UAEC) 与黄体 (Lut) NP-UAEC 相比经历了适应；即在新分离的 UA 中看到的反应 内皮维持在许多通道上。其中包括 ERK 1/2 -MAPK 与 eNOS 激活的偶联 大量 ATP 刺激的 NO 产生 (P > Fol-NP> Lut-NP) 以及雌激素可能改变 Ca2+ 的作用 介导的NO。剪切应力和/或雌激素都是 NO 合成和/或 eNOS 的非常有效的刺激剂 表达并共同引发 P-UAEC 中 ERK 1/2 MAPK 信号通路的协同激活。 血流的升高也会大大增加层流剪切应力。其他信号通路（例如 PI3 激酶）可能在体内情况中发挥额外的收敛作用。总体假设：生理状态 高 DBF/剪切应力和升高的雌激素（例如 P 和 Fol）导致 UAEC 的“程序性适应” 调节与 NO 产生相关的重要内皮功能。具体强调的是 增加 eNOS 激活、eNOS 表达、UAEC 信号激酶（例如 ERK 1/2 和/或 PIS）的机制 激酶）和细胞间通讯（以同步 Ca2+ 爆发来测量）。 目标 I-生理模型：使用来自孕妇的 V/Vo 衍生的新鲜分离的 UA 内皮细胞（第 0 代）， 将进行骑自行车（Foland Lut）和卵巢切除（车辆与雌激素处理）羊的研究 评估雌激素改变基础、ATP 和龙霉素的细胞调节（非基因组和基因组） 通过直接成像细胞内 NO 生成与 [Ca2+]i/Ca2+ 爆发同时进行 eNOS 激活 评估 ER 和汇聚信号中间体（ERK1/2 MAPK 和/或 PIS 激酶）途径的作用。 目标 II - 剪切应力模型：长期层流剪切应力 [0（静态）、3 和 15 达因/cm2] 的影响 没有E2（3关于长期培养的“程序化UAEC适应”（第4-5代）Lut NP-UAEC，Fol NP- UAEC 和 P-UAEC。从我们关注的时间隔离中“编程”的具体适应标记 包括：a) eNOS 表达和 ATP/lonomycin 刺激的 eNOS 激活和 [Ca2+]i/Ca2+ 爆发和 b) ATP/lonomycin 刺激 ERK1/2 和 AKT 磷酸化；以及 ERK-MAPK 和/或 PIS 激酶的抑制 介导刺激 NO 产生和 [Ca2+]i/Ca2+ 爆发。我们假设包含剪切应力 雌激素对于恢复特定标记和“编程”到的反应幅度是必要的 怀孕期间和目标 I 中定义的卵泡期通过高流量和雌激素产生的 UAEC。数据如下 推导将为理解剪切应力和剪切应力之间的相互作用提供第一个机械框架 雌激素调节 UAEC NO 和聚合信号机制。我们最近的初步数据 表明在并发先兆子痫的妊娠中，子宫动脉剪切应力确实降低了。 因此，这些机制在有或没有 IUGR 的先兆子痫期间也可能出现功能障碍。