Control of Microcirculatory Exchange Function

描述（由申请方提供）：微血管通透性增加是炎症的标志。以前的研究集中在如何防止炎症条件下的发病和维持高渗透性。.我们建议测试的中心假设，微血管通透性调节平衡信号机制，保持体内平衡，通过提高渗透性或屏障性能，分别。我们将研究eNOS易位作为刺激高渗透性的机制和在早期炎症阶段抑制高渗透性的机制。我们将在内皮细胞和横纹肌中研究这些过程。我们假设eNOS易位是必要的，以实现有效的NO浓度或附近的渗透性效应。我们还假设，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微血管壁的生理屏障特性的过程。我们进一步提出，小GTP结合蛋白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的是要测试：具体的假设1。eNOS易位到胞质（非高尔基体胞质）是微血管通透性内皮调节的一个步骤。具体目标1.探讨内皮型一氧化氮合酶（eNOS）转位与内皮和微血管通透性调节的关系。特异性假设2：cAMP通过Epac使激动剂诱导的高通透性失活或逆转。具体目的2.1研究高通透性的定时失活/逆转。具体目标2.1.确定刺激Epac是否逆转（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方法来阐明假设的机制。公共卫生相关性：更好地理解分子机制和高通透性与其在炎症过程中的“生理”失活或逆转之间的时间，应该为促进高通透性的失活或逆转提供机会之窗，以预防（微）血管疾病中的并发症，如过度水肿和筋膜室综合征。

DESCRIPTION (provided by applicant): 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 mechanisms. PUBLIC HEALTH RELEVANCE: A better understanding of the molecular mechanisms and of the timing between hyperpermeability and its "physiological" inactivation or reversal in inflammatory processes should provide a window of opportunity for promoting inactivation or reversal of hyperpermeability to prevent complications such as excessive edema and compartment syndrome in (micro)vascular disease.