PREVENTION OF INCREASED VASCULAR PERMEABILITY

预防血管通透性增加

• 批准号: 3355345
• 负责人: FRED L MINNEAR
• 金额: $ 8.37万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3355345
• 关键词: beta adrenergic agent; cardiovascular pharmacology; cell morphology; cyclic AMP; endotoxins; fluorescence microscopy; histamine; isoproterenol; microtubules; phase contrast microscopy; pulmonary edema; respiratory disorder chemotherapy; sheep; terbutaline; thrombin; tissue /cell culture; vascular endothelium permeability

Beta-adrenergic agonists such as isoproterenol and terbutaline have been shown to attenuate the edema that various agents can produce in systemic and pulmonary vascular beds. We hypothesize that beta-agonists can prevent and possibly reverse the development of edema by a permeability-decreasing effect on vascular endothelial cells as well as hemodynamic effects and that this effect is unique to beta-agonists as opposed to other vasodilator substances. Furthermore, we hypothesize that this permeability-decreasing effect is mediated via cyclic AMP which alters the intracellular microfilaments and/or microtubules in such a way as to change the shape of the cell. This proposal will utilize in vitro and in vivo experimentation to demonstrate whether beta-agonists have a permeability-decreasing effect and to elucidate the mechanism for this activation the in vitro studies will utilize cultured endothelial cells from arteries. These studies will determine if beta-agonists can prevent and reverse the increased endothelial permeability produced by various permeability-increasing mediators such as thrombin, endotoxin, and histamine. The measurement for endothelial permeability will be the clearance of 125I-albumin across the cultured endothelial cell monolayers seeded on Nucleopore (0.8 um) filters. Radioligand binding studies will be done to demonstrate the presence of beta-receptors on the cultured endothelial cells. The in vitro studies will also determine if the permeability-increasing agents alter cell shape and the number and organization of cellular microfilaments and microtubules and if beta-agonists can prevent these alterations. Cell shape and microfilament changes will be assessed by time-lapse video, phase contrast and immunofluorescent microscopy. The proposed studies will focus on the pulmonary endothelium. With the information gained by the in vitro experimentation, in vivo studies, utilizing the chronic sheep lung lymph fistula preparation, will be done to differentiate between the effects of beta-agonists on vascular permeability and hemodynamics in the awake animal. These in vivo sheep studies will also focus on the pulmonary vasculature. In parallel with these objectives is the long-term objective of determining the transcellular signal pathways that may alter the shape of endothelial and epithelial cells and influence their function as semipermeable membranes.

β-肾上腺素能激动剂，例如异丙肾上腺素和特布他林 已被证明可以减轻各种药物可以减轻的水肿 产生于全身和肺血管床。 我们假设 β-激动剂可以预防并可能逆转 通透性降低作用导致水肿的发展 血管内皮细胞以及血流动力学效应和 与其他药物相比，这种效应是 β 激动剂所独有的 血管扩张物质。 此外，我们假设这 通透性降低作用是通过环 AMP 介导的， 改变细胞内的微丝和/或微管 这种方法可以改变细胞的形状。 该提案将 利用体外和体内实验来证明 β-激动剂是否具有通透性降低作用以及 为了阐明这种激活的机制，进行了体外研究 将利用来自动脉的培养内皮细胞。 这些研究 将确定β受体激动剂是否可以预防和逆转 各种因素引起的内皮通透性增加 增加通透性的介质，如凝血酶、内毒素、 和组胺。 内皮通透性的测量 将是 125I-白蛋白在培养物中的清除率 内皮细胞单层接种在 Nucleopore (0.8 um) 过滤器上。 将进行放射性配体结合研究以证明 培养的内皮细胞上存在β受体。 这 体外研究还将确定通透性增加是否 药剂改变细胞形状以及细胞的数量和组织 细胞微丝和微管，如果β-激动剂可以 防止这些改变。 细胞形状和微丝变化 将通过延时视频、相差和 免疫荧光显微镜。 拟议的研究将集中于 在肺内皮上。 随着所获得的信息 体外实验，体内研究，利用慢性 羊肺淋巴瘘的准备，将做鉴别 β-激动剂对血管通透性的影响和 清醒动物的血流动力学。 这些绵羊体内研究 还将重点关注肺血管系统。 并行于 这些目标是确定的长期目标 可能改变形状的跨细胞信号通路 内皮细胞和上皮细胞并影响它们的功能 半透膜。