Control of Microcirculatory Exchange Function

摘要 微血管通透性增加是炎症的标志。先前的研究 重点是如何防止发病和维持下的渗透性升高 炎性条件。.我们建议测试中心假设，微血管 渗透性由维持体内平衡的信号传导机制调节 通过分别增强渗透性或阻隔性。我们会调查eNOS 易位作为刺激高渗透性的机制， 在早期炎症阶段出现血管通透性增高。我们将研究这些过程， 内皮细胞和横纹肌。我们假设eNOS转位是必要的， 以在渗透性效应器处或附近获得有效的NO浓度。我们也 假设在或接近高渗透性反应的峰值时，内皮（或 组织）开始抑制或逆转高渗透性并恢复生理性的过程。 微血管壁的屏障特性。我们进一步提出，小GTP结合 蛋白质Epac/Rap-1作为“屏障增强因子”并参与 超渗透性-失活/逆转过程。具体的假设和具体的目标， 具体的假设是：1。eNOS易位至胞质溶胶（非高尔基体胞质溶胶 隔室）是微血管通透性的内皮调节中的一个步骤。具体目标 1.探讨内皮型一氧化氮合酶（eNOS）转位与血管内皮细胞功能调节的关系， 微血管通透性特异性假说2：cAMP通过Epac失活或逆转 激动剂诱导的高通透性。具体目标2.1调查 高渗透性的失活/逆转。具体目标2.1.以确定是否 Epac的刺激逆转（灭活）激动剂诱导的高通透性。我们将应用 活体显微镜结合计算机辅助图像分析和分子生物学 生物学方法来阐明假设的机制。

ABSTRACT Increased microvascular permeability is a hallmark of inflammation. Previous research has focused on how to prevent the onset and the maintenance of the elevated permeability under inflammatory conditions. . We propose to test the central hypothesis that Microvascular permeability is regulated by counterbalancing signaling mechanisms that maintain homeostasis by enhancing permeability or barrier properties, respectively. We will investigate eNOS translocation as a mechanism that stimulates hyperpermeability and mechanisms that inactivate hyperpermeability in the early inflammatory phase. We will study these processes in endothelial cells and in striated muscle. We hypothesize that eNOS translocation is necessary to achieve effective NO concentrations at or near the effectors for permeability. We also hypothesize that at or near the peak of the hyperpermeability response, the endothelium (or tissue) begins a process to inactivate or reverse hyperpermeability and restore the physiological barrier properties of the microvascular wall. We further propose that the small GTP-binding proteins Epac/Rap-1 serve as `Barrier Enhancing Factors' and participate in the hyperpermeability-inactivation/reversal process. The Specific Hypotheses and Specific Aims to be tested are: SPECIFIC HYPOTHESIS 1. eNOS translocation to cytosol (non-Golgi cytosol compartment) is a step in endothelial regulation of microvascular permeability. SPECIFIC AIM 1. To investigate the relationship between eNOS translocation and regulation of endothelial and microvascular permeability. SPECIFIC HYPOTHESIS 2: cAMP via Epac inactivates or reverses agonist-induced hyperpermeability. SPECIFIC AIM 2.1 To investigate the timed inactivation/reversal of hyperpermeability. SPECIFIC AIM 2.1. To determine whether or not stimulation of Epac reverses (inactivates) agonist-induced hyperpermeability. We will apply intravital microscopy in conjunction with computer-assisted image analysis and molecular biology approaches to elucidate the postulated mechanims.