MODULATION OF LUNG ENDOTHELIAL PERMEABILITY BY AUTACOIDS

Autacoids 对肺内皮细胞通透性的调节

• 批准号: 3357789
• 负责人: Frederick R Haselton
• 金额: $ 11.01万
• 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/3357789
• 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.

肺中血液组织渗透性的测量具有 被证明是非常困难的。 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 来自肺动脉、静脉和微血管的细胞层。 的 方法提供了一种新的方法，并克服了许多 用于研究增加的整个动物方法的困难 渗透性水肿通常是成人呼吸道疾病的特征， 痛苦综合症 血液组织渗透性增加 难以在渗透性增加的动物模型中证明 水肿 例如，研究人员有这样的羊肺- 淋巴制剂和刺激物如组胺、内毒素， 凝血酶作为肺损伤模型的相似性增加 渗透性水肿 然而，血液组织渗透性的变化 在这些动物模型中不能清楚地识别，因为肺 损伤伴随血管内压力增加， 也可以解释观察到的流体流量增加， 溶质。 体外方法将有助于确定特定的渗透性 在动物中产生肺损伤的药剂引起的变化。 使用 细胞培养方法的最新进展， 血液组织内皮成分的特征 障碍已经开始调查。 然而，这些结果是 受到质疑，因为这些体外方法发现渗透性 比体内估计值高1000倍。 在初步实验中，我们的体外方法发现内皮细胞 渗透性与体内估计相似。 这种新方法 利用在微载体上培养的单层内皮细胞 微珠作为选择性屏障来控制分子通过 来自液相色谱柱的移动的相的物质 到细胞覆盖的微载体内的固定相 珠 我们建议使用这种方法来阐明 血液组织渗透性的内皮屏障，此外， 测量渗透性如何被自然改变 发生的刺激，即autacoids。 该提案有三个主要内容 目的：1）测定体外培养的人脐静脉内皮细胞的细胞间通透性， 肺内皮细胞层对甘露醇和聚乙烯 乙二醇; 2）表征肺的渗透性 内皮细胞层相对于示踪剂大小和电荷的变化;和 3)为了检验自伤药改变肺功能的假设， 通过测量渗透性变化的内皮细胞渗透性 由三种autacoids产生，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。