ENDOTHELIUM-DERIVED VASODILATORS IN PREGNANCY

妊娠期内皮源性血管扩张剂

• 批准号: 2225318
• 负责人: RONALD R MAGNESS
• 金额: $ 15.4万
• 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 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2225318
• 关键词: adenylate cyclase; angiotensin II; cyclic AMP; cyclic GMP; enzyme activity; estrogens; gel electrophoresis; gene expression; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; nitric oxide; northern blottings; pregnancy; progesterone; prostacyclins; prostaglandin endoperoxide synthase; radiotracer; sheep; vascular smooth muscle; vasodilators; western blottings

Normal cardiovascular adaptations in pregnancy include generalized vasodilation and reduced uterine and systemic vasoconstriction in response to infused angiotensin II (ANG II) and alpha-agonists. During human and ovine pregnancy the uterine vascular bed is even less reactive to vasoconstriction by ANG II compared to systemic vasculature. The long-term goals of this research project are to determine if the vasodilatory endothelial factors prostacyclin (PGI2) and endothelium- derived relaxing factor/nitric oxide (EDRF/NO) are responsible for modulating maternal vascular smooth muscle (VSM) function in normal pregnancy and if estrogen and/or progesterone, steroid hormones elevated in pregnancy, will modulate their productions. We know from in vitro models which evaluate endothelial/VSM interactions that endothelium- derived PGI2 and EDRF/NO are released in a paracrine fashion to modulate production of the second messenger cyclic nucleotides cAMP and cGMP, respectively, which then decrease VSM constriction and tone. The overall hypothesis of this project is that uterine and systemic endothelium- derived PGI2 and EDRF/NO productions are increased in normal pregnancy and that they modulate VSM cAMP and cGMP to control dilation and arterial responsiveness to vasoconstrictors in particular ANG II. The specific aims of this project are to define the molecular and enzymatic mechanisms controlling PGI2 and EDRF/NO production in normal pregnancy in order to further understand uterine and systemic artery endothelial modulation of VSM cAMP and cGMP production. We will evaluate adenylate and guanylate cyclase enzyme sensitivity in VSM from pregnant and nonpregnant ewes. Also planned is the delineation of which enzymes are activated in association with increased production of endothelium-derived PGI2 and EDRF/NO during pregnancy; cyclooxygenase (PGHS-1 & PGHS-2), PGI2 synthase and EDRF/NO synthase will be studied using Western immunoblots, immunocytochemistry and productions of PGI2, cAMP, and cGMP; specific activity studies also are planned. Experiments are proposed to study the gene regulation of cyclooxygenase, PGI2 synthase EDRF/NO synthase in pregnancy using Northern blots and in situ hybridization. The direct effects of estrogen and/or progesterone to regulate PGI2 and EDRF/NO production and gene expression will provide mechanisms by which placental/ovarian steroid production modifies VSM function in pregnancy. Addressing these aims will increase our knowledge concerning the enzymatic and molecular control endothelial/VSM interactions associated with normal and potentially pathologic cardiovascular adaptations in pregnancy.

妊娠期正常的心血管适应包括全身性的 血管舒张并减少子宫和全身血管收缩 对输注血管紧张素 II (ANG II) 和 α 激动剂的反应。 期间 人类和绵羊怀孕时，子宫血管床的反应性更小 与全身脉管系统相比，ANG II 的血管收缩作用。 这 该研究项目的长期目标是确定是否 血管舒张内皮因子前列环素 (PGI2) 和内皮细胞- 衍生松弛因子/一氧化氮 (EDRF/NO) 负责 调节正常情况下母体血管平滑肌 (VSM) 功能 怀孕以及雌激素和/或黄体酮、类固醇激素升高 在怀孕期间，会调节它们的产生。 我们从体外得知 评估内皮/VSM 相互作用的模型 衍生的 PGI2 和 EDRF/NO 以旁分泌方式释放以调节 第二信使环核苷酸cAMP和cGMP的产生， 分别降低 VSM 收缩和音调。 整体 该项目的假设是子宫和全身内皮细胞 正常妊娠时衍生的 PGI2 和 EDRF/NO 产量增加 他们调节 VSM cAMP 和 cGMP 来控制扩张和动脉 对血管收缩剂特别是 ANG II 的反应。 具体的 该项目的目的是定义分子和酶机制 控制正常妊娠中 PGI2 和 EDRF/NO 的产生，以便 进一步了解子宫和全身动脉内皮的调节 VSM cAMP 和 cGMP 生产。 我们将评估腺苷酸和鸟苷酸 怀孕和非怀孕母羊 VSM 中环化酶的敏感性。 还计划描述哪些酶在 与内皮源性 PGI2 产生增加相关 怀孕期间的 EDRF/NO；环氧合酶（PGHS-1 和 PGHS-2）、PGI2 合酶 EDRF/NO合酶将使用Western免疫印迹进行研究， 免疫细胞化学和 PGI2、cAMP 和 cGMP 的产生；具体的 还计划开展活动研究。 建议进行实验来研究 环氧合酶、PGI2合酶、EDRF/NO合酶的基因调控 使用 Northern 印迹和原位杂交进行妊娠。 直接的 雌激素和/或孕激素调节 PGI2 和 EDRF/NO 的作用 生产和基因表达将提供机制 胎盘/卵巢类固醇的产生会改变怀孕期间的 VSM 功能。 解决这些目标将增加我们对以下方面的认识： 酶促和分子控制内皮/VSM 相互作用相关 具有正常和潜在病理性心血管适应 怀孕。