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Shear Stress-mediated Pregnancy Adaptations of Uterine Artery Endothelial...

剪切应力介导的子宫动脉内皮细胞的妊娠适应...

基本信息

批准号：
7189523
负责人：
RONALD R MAGNESS
金额：
$ 17.19万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-05-01 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7189523
关键词：
Address Agonist Arteries Birth Weight Blood Vessels Blood flow Cell Communication Cell physiology Cells Connexin 43 Coupled Coupling Cultured Cells Cytosolic Phospholipase A2 Data Disease Elevation Employee Strikes Endothelial Cells Endothelium Estrogen Receptors Estrogens Fetal Growth Fetal Growth Retardation Functional disorder Gap Junctions HTATIP gene ICI 182780 In Vitro Link Luteal Phase MAP Kinase Gene MAPK Signaling Pathway Pathway Measures Mediating Microscope Mitogen Activated Protein Kinase 1 Modeling Neonatal Nitric Oxide PLA2G4A gene Pathologic Patient currently pregnant Perfusion Phenotype Phosphorylation Phosphotransferases Pre-Eclampsia Pregnancy Production Program Development Proteins Reporting Research Personnel Role Sheep Signal Transduction Slide Stimulus Testing Third Pregnancy Trimester Time Vascular Endothelial Cell Vascular Endothelial Growth Factor Receptor-1 Vascular Endothelial Growth Factor Receptor-2 Vascular Endothelial Growth Factors Vasodilation Vasodilation disorder Vasodilator Agents angiogenesis base cyclooxygenase 1 fetal human PLA2G4A protein human embryonic stem cell in vivo neovascularization novel programs receptor response shear stress

项目摘要

Pregnancy is associated with dramatic increases in uterine blood flows correlating with fetal growth and survival. The uteroplacental unit has prominent production of estrogen, NO and eNOS expression. Estrogen increases NO production. Uterine Artery Endothelial Cells (UAEC) of Pregnancy (P) show specific "Programmed Adaptation" [not seen in Nonpregnant (NP)-UAEC] that maintains in cell culture passages. "Programmed Adaptation" thus is a UAEC response of freshly isolated UA Endothelium ex vivo, that upon many passages does not diminish. The overall hypothesis of this proposal is that: In pregnancy, shear stress/flow in the presence of estrogen causes "Programmed Adaptation" of the UAEC to modulate important endothelial functions associated with NO-mediated vasodilator production. We will give specific emphasis to those mechanisms that increase eNOS activation, eNOS expression, UAEC signaling kinases (e.g. ERK 1/2),and cell-cell communication (measured as synchronized Ca2+ bursts) as modulated by VEGF and Gapjunctions. Aim 1: Effects of prolonged Laminar Shear Stress with and without E2(3on expression of key functional proteins (eNOS, COX-1, PGIS, cPLA2, CX43, ERa, ERp, VEGFR-1, VEGFR-2, and NP-1) necessary for adaptation in UAECs from Luteal Phase Nonpregnant Sheep (NP-UAEC) and Late Pregnant Sheep (P- UAEC). Aim 2: Effects of prolonged Laminar Shear Stress with and without E2pon "Programmed Adaptation" in UAECs from NP-UAEC and P-UAEC. Three "Programmed Adaptation Specific Markers" include: a)ATP/VEGF- stimulated eNOS activation; b) ATP/VEGF-stimulated ERK1/2 phosphorylation; and c) ERK-MAPK and Gap junction mediated ATP-induced Ca2+ bursts. Aim 3: Effects of Estrogen Receptor antagonism with ICI 182,780 on the expression of key functional proteins (as in Aim 1) necessary for Shear Stress/E2p adaptation of UAECs. Aim 4: Effects of Estrogen Receptor antagonism with ICI 182,780 on Shear Stress/E2p UAEC "Programmed Adaptation Specific Markers" (as in Aim 2). Aim 5: Evaluate the effect of prolonged Laminar Shear Stress on development of programmed endothelial cell functions comparing responses of endothelial cells derived from human Embryonic Stem Cells (Project III) to human Embryonic Stem Cells (Project IV; Core B) themselves. These studies will provide a mechanistic framework for understanding interactions between shear stress and estrogen to regulate NO production via VEGF and gap junction mediated cell-cell communication. Vascular adaptations in pregnancy are important because increases in fetoplacental/uteroplacental perfusion, are linked directly to fetal growth and these mechanisms are dysfunctionalin pathologic pregnancies (e.g. preeclampsia/ IUGR).

妊娠与子宫血流量的急剧增加有关，子宫血流量与胎儿的生长和存活有关。子宫胎盘单位具有显著的雌激素产生、NO和eNOS表达。雌激素增加NO 生产妊娠期子宫动脉内皮细胞（UAEC）具有特异性的“程序性适应” [not在非妊娠（NP）-UAEC中观察到]，其维持在细胞培养传代中。因此，“程序化适应”是一种新鲜分离的UA内皮素离体的UAEC反应，在多次传代后不会减少。的这一建议的总体假设是：在怀孕期间，在雌激素存在下的剪切应力/流动引起UAEC的“程序性适应”，以调节重要的内皮功能，一氧化氮介导的血管扩张剂的产生。我们将特别强调那些机制，增加eNOS活化、eNOS表达、UAEC信号传导激酶（例如ERK 1/2）和细胞-细胞图2示出了由VEGF和缝隙连接调节的细胞间通讯（测量为同步的Ca 2+爆发）。目的1：长时间的层流剪切应力（含和不含E2 β）对关键功能蛋白表达的影响蛋白质（eNOS、考克斯-1、PGIS、cPLA 2、CX 43、ER α、ER β、VEGFR-1、VEGFR-2和NP-1），来自黄体期非妊娠绵羊（NP-UAEC）和晚孕绵羊（P-UAEC）的UAEC的适应性 UAEC）。目的2：在有和没有E2的情况下，延长的层流剪切应力对“程序化适应”的影响，来自NP-UAEC和P-UAEC的UAEC。三种“程序化适应特异性标志物”包括：刺激的eNOS活化; B）ATP/VEGF刺激的ERK 1/2磷酸化;和c）ERK-MAPK和Gap 连接介导的ATP诱导的Ca 2+爆发。目的3：用ICI 182，780拮抗雌激素受体对关键功能性细胞因子表达的影响。 UAEC的剪切应力/E2 p适应所必需的蛋白质（如目的1中所述）。目的4：ICI 182，780对剪切应力/E2 p UAEC的雌激素受体拮抗作用 “程序化适应特定标记”（如目标2）。目的5：评价长期层流剪切应力对程序性内皮细胞发育的影响。细胞功能比较人胚胎干细胞来源的内皮细胞的反应（项目III）到人类胚胎干细胞（项目IV;核心B）本身。这些研究将为理解剪切应力和剪切力之间的相互作用提供一个机制框架。雌激素通过VEGF和缝隙连接介导的细胞间通讯调节NO的产生。血管妊娠期的适应是重要的，因为胎儿胎盘/子宫胎盘灌注的增加，直接影响胎儿生长，这些机制在病理性妊娠（如先兆子痫/ IUGR）中功能障碍。