MODULATION OF LUNG ENDOTHELIAL PERMEABILITY BY AUTACOIDS

Autacoids 对肺内皮细胞通透性的调节

• 批准号: 3357788
• 负责人: Frederick R Haselton
• 金额: $ 10.79万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-10 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3357788
• 关键词: adult respiratory distress syndrome; blood pressure; chromatography; endotoxins; ethylene glycols; histamine; hormones; lung disorder; mannitol; norepinephrine; polyethylenes; solute; thrombin; tissue /cell culture; vascular endothelium permeability

The measurement of blood-tissue permeability in the lung has proven to be extremely difficult. We have developed a new in vitro method to measure the permeability of cultured endothelial cell layers from pulmonary arteries, veins, and microvessels. The method offers a fresh approach and overcomes many of the difficulties of whole animal methods used to study the increased permeability edema often characteristic of the adult respiratory distress syndrome. Blood-tissue permeability increases are difficult to prove in animal models of increased permeability edema. For example, investigators have such the sheep lung- lymph preparation and stimuli such as histamine, endotoxin, and thrombin as models of lung injury similar to increased permeability edema. However, blood-tissue permeability changes cannot be clearly identified in these animal models, since the lung injury is accompanied by increased intravascular pressures which could also account for the observed increased flux of fluid and solutes. In vitro methods will help identify specific permeability changes induced by agents producing lung injury in animals. Using recent advances in cell culture methods, the permeability characteristics of the endothelial component of the blood-tissue barrier have begun to be investigated. However, these results are being questioned, since these in vitro methods find permeabilities 1000 times greater than in vivo estimates. In preliminary experiments, our in vitro method finds endothelial permeabilities similar to in vivo estimates. This new method utilizes a monolayer of endothelial cells cultured on microcarrier beads as a selective barrier to control the passage of molecular species from the mobile phase of a liquid chromatography column to the stationary phase within the cell covered microcarrier beads. We propose to use this method to clarify the role of the endothelial barrier in blood-tissue permeability and, in addition, measure how the permeability properties are altered by naturally occurring stimuli, i.e. autacoids. This proposal has three main aims: 1) to measure the in vitro intercellular permeability of the pulmonary endothelial cell layer to mannitol and polyethylene glycol; 2) to characterize the permeability of the pulmonary endothelial cell layer with respect to tracer size and charge; and 3) to test the hypothesis that autacoids alter pulmonary endothelial cell permeability by measuring permeability changes produced by three autacoids, which have been described as reversible modulators of blood-tissue permeability in the lung: norepinephrine, thrombin and histamine.

肺内血液组织通透性的测量有 事实证明，这是非常困难的。我们开发了一种新的 体外测定培养内皮细胞通透性的方法 肺动、静脉和微血管的细胞层。这个 方法提供了一种新的方法，并克服了许多 用全动物方法研究增加的难度 通透性水肿通常是成人呼吸系统的特征 窘迫综合症。血液组织通透性增加是 在通透性增加的动物模型中难以证明 浮肿。例如，调查人员有这样一种绵羊肺- 淋巴准备和刺激，如组胺、内毒素和 凝血酶作为肺损伤模型类似于增生性肺损伤 通透性水肿。然而，血液组织的通透性改变 在这些动物模型中无法清楚地识别，因为肺 损伤伴随着血管内压的增加 也可以解释观察到的流体流量的增加和 溶质。体外方法将有助于识别特定的渗透性 引起动物肺损伤的因素引起的变化。vbl.使用 细胞培养方法的最新进展--渗透性 血液组织内皮细胞成分的特性 障碍已经开始被调查。然而，这些结果是 受到质疑，因为这些体外方法发现通透性 比体内估计的要大1000倍。 在初步实验中，我们的体外方法发现内皮细胞 渗透率与体内估计的类似。这种新方法 利用在微载体上培养的单层内皮细胞 珠子作为控制分子通过的选择性屏障 从液体色谱柱的流动相中分离出的物种 到细胞覆盖的微载体内的固定相 珠子。我们建议使用这种方法来澄清 内皮屏障在血液组织通透性中的作用，此外， 测量渗透性如何在自然条件下发生改变 发生的刺激，即眼镜蛇。这项建议主要有三个方面 目的：1)测定体外细胞间通透性。 肺内皮细胞层对甘露醇和聚乙烯的影响 乙二醇法；2)表征肺组织通透性 关于示踪剂大小和电荷的内皮细胞层；以及 3)检验耳廓改变肺脏的假说 通过测量内皮细胞通透性的变化来确定通透性 由三个金刚类制作，它们被描述为 血液组织通透性的可逆调节剂 肺：去甲肾上腺素、凝血酶和组胺。