PREVENTION OF INCREASED VASCULAR PERMEABILITY

预防血管通透性增加

• 批准号: 3355342
• 负责人: FRED L MINNEAR
• 金额: $ 8.5万
• 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 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3355342
• 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，如异丙肾上腺素和特布他林 已经被证明可以减轻各种药剂可以缓解的浮肿 在全身和肺血管床中产生。我们假设 β-激动剂可以预防并可能逆转 通透性降低引起的水肿 血管内皮细胞以及血流动力学效应和 这种效应是β-激动剂独有的，而不是其他 血管扩张剂物质。此外，我们假设这一点 降低通透性的作用是通过环状AMP介导的 改变细胞内微丝和/或微管 一种改变细胞形状的方法。这项提议将 利用体外和体内实验证明 β-激动剂是否具有降低通透性的作用 为了阐明这种激活的机制，体外研究 将利用培养的动脉内皮细胞。这些研究 将确定β-激动剂是否可以预防和逆转 由多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、 还有组胺。血管内皮细胞通透性测定 将~(125)I-白蛋白清除到培养的 种植在核孔(0.8微米)滤膜上的内皮细胞单层。 将进行放射性配基结合研究，以证明 培养的内皮细胞上有β受体的存在。这个 体外研究也将确定通透性增加是否 代理人改变细胞的形状和数量和组织 细胞微丝和微管，如果β-激动剂可以 防止这些改动。细胞形态和微丝变化 将通过延时视频、相位对比度和 免疫荧光显微镜。拟议的研究将集中于 在肺内皮细胞上。通过以下方式获得的信息 体外实验，体内研究，利用慢性 羊肺淋巴瘘的准备，将做好鉴别 β-受体激动剂对血管通透性的影响与 清醒动物的血流动力学。这些活体绵羊研究 还将关注肺血管系统。与...并行 这些目标是确定 可能改变细胞形态的跨细胞信号通路 内皮细胞和上皮细胞，并影响它们的功能 半透膜。