PREVENTION OF INCREASED VASCULAR PERMEABILITY

预防血管通透性增加

• 批准号: 3355347
• 负责人: FRED L MINNEAR
• 金额: $ 12.73万
• 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 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3355347
• 关键词: adenylate cyclase; albumins; beta adrenergic agent; biological signal transduction; body water; calcium; calcium indicator; cardiovascular pharmacology; cell morphology; cyclic AMP; cyclic GMP; drug administration routes; electron microscopy; guinea pigs; hemodynamics; immunofluorescence technique; inositol phosphates; intercellular connection; membrane model; microfilaments; nitric oxide; nonhuman therapy evaluation; pulmonary edema; respiratory disorder chemotherapy; second messengers; sheep; thrombin; tissue /cell culture; vascular endothelium permeability; vasodilators

Beta-adrenergic agonists have been shown to attenuate the edema and increased vascular permeability that various agents can produce in systemic and pulmonary vascular beds. The objectives of this renewal are to: (1) determine if beta-agonists reduce water and protein fluxes by a direct effect on endothelial permeability of water and albumin and/or by changes in hemodynamics; (2) determine the validity of beta-agonists as potential therapeutic agents for pulmonary edema and increased vascular permeability, and (3) determine the mechanism, from stimulation of adenylate cyclase to cytoskeletal and junctional organization, for enhancement of the barrier function of the endothelium. We hypothesize that beta-agonists, especially when administered as an aerosol, can prevent or reduce increased fluid and protein fluxes with minimal alterations in hemodynamics, gas exchange, and lung compliance. We also hypothesize that these agents modify permeability by their action on junctional associated microfilaments to prevent alterations in junctional architecture, cell shape, and permeability. The overall design of this renewal utilizes a structure-function approach and models of cell culture, isolated organ, and whole animal. These models focus on the pulmonary endothelium and utilize albumin uptake as the common indicator of protein permeability. Measurements of capillary pressure, hydraulic conductivity, diffusive albumin flux, and the albumin reflection coefficient in the isolated, perfused guinea pig lung will be used to differentiate between hemodynamic and direct permeability effects of beta- agonists administered via the circulation or airway. Measurements of inositol phosphate metabolism and cytosolic calcium will be made to determine if beta-agonists affect these processes in opposition to the permeability - increasing action of alpha-thrombin. Sheep will be used to determine if beta-agonists are potential therapeutic agents for pulmonary edema and increased vascular permeability, the latter assessed by the calculation of the reflection coefficient. Studies will compare intravenous versus airway administration of the agents on fluid and protein fluxes, hemodynamics, gas exchange, and lung compliance. Endothelial cell monolayers cultured from bovine pulmonary artery endothelial cells will be used to continue to determine the mechanism for the permeability - decreasing action of beta-agonists. Drugs designed to activate or inhibit membrane cyclases and kinases will also determine if the action of cAMP is shared by cGMP-enhancing agents. Correlative light, immunofluorescence, and electron microscopy will determine if beta-agonists increase the number and interaction of the peripheral band of actin microfilaments with junctional elements of endothelial cells and, thereby, prevent the alterations in the endothelial junction, in cell shape, and in permeability induced by alpha-thrombin. In parallel with these objectives is the long- term goal of determining the transcellular signalling pathways that may alter the shape of endothelial and epithelial cells to influence their function as semipermeable membranes.

β-肾上腺素能激动剂已被证明可以减轻水肿和 血管通透性增加的各种因素可产生在全身 和肺血管床。这次更新的目标是：(1) 确定β-受体激动剂是否通过直接的 水、白蛋白和/或改变对内皮通透性的影响 在血流动力学中；(2)确定β-激动剂的有效性 治疗肺水肿和血管通透性增加的药物， 以及(3)从腺苷环化酶的刺激作用到 细胞骨架和连接组织，以增强屏障 内皮细胞的功能。我们假设β-激动剂，尤其是 当作为气雾剂使用时，可以防止或减少液体增加和 在血流动力学、气体交换和 肺顺应性。我们还假设这些药物可以改变通透性。 通过它们对连接相关微丝的作用来防止 连接结构、细胞形状和渗透性的变化。这个 本次更新的总体设计采用了结构-功能相结合的方法 细胞培养、分离器官和整个动物的模型。这些型号 关注肺内皮细胞，利用白蛋白摄取作为常见的 蛋白质通透性的指标。毛细管压力的测量， 水力传导性、扩散白蛋白通量和白蛋白反射 分离、灌流的豚鼠肺中的系数将用于 区分β-受体阻滞剂的血流动力学效应和直接通透性效应 通过循环或呼吸道给予的激动剂。测量 肌醇磷酸代谢和胞浆钙将被用来 确定β-激动剂是否影响这些过程而不是 α-凝血酶的通透性增加作用。绵羊将习惯于 确定β-激动剂是否是治疗肺部疾病的潜在药物 水肿和血管通透性增加，后者由 反射系数的计算。研究将比较 静脉给药与呼吸道给药对液体和蛋白质的影响 流量、血流动力学、气体交换和肺顺应性。内皮细胞 从牛肺动脉内皮细胞培养的单层将 用来继续确定渗透性的机制- 降低β-受体激动剂的作用。旨在激活或抑制的药物 膜周期酶和激酶也将决定cAMP的作用是否 由cGMP增强型代理共享。相关光，免疫荧光， 而电子显微镜将确定β-激动剂是否会增加这种数量。 以及肌动蛋白微丝的外周带与 内皮细胞的连接元件，从而防止 内皮连接、细胞形状和通透性的改变 由α-凝血酶诱导。与这些目标并行的是长期的- 确定跨细胞信号通路的长期目标 改变内皮细胞和上皮细胞的形状以影响它们的 起到半透膜的作用。