Clinical and Experimental Therapeutics, College of Pharmacy, University of Georgia, Augusta, GA, USA.
Methods Mol Biol. 2021;2367:165-175. doi: 10.1007/7651_2020_310.
Whereas physiological vascular permeability (VP) mediates selective transport of plasma, electrolytes, proteins, and cells across an intact endothelial barrier, pathological VP results in the loss of endothelial barrier integrity. Whereas physiological VP is a feature of regular host defense and tissue repair, compromised barrier function may lead to aberrant vascular leakage, concurrent tissue edema, and inflammation eventually causing life-threatening conditions such as acute lung injury or acute respiratory distress syndrome, cancer, kidney injury, etc. Measurement of VP helps to identify, design, and optimize anti-leak therapies. Further, it can define the effect of a stimulus or a gene modulation in endothelial-barrier regulation. The degree of VP can be of importance to determine the stage of cancer and disease prognosis. This chapter discusses Miles assay, which is a well-established, relatively simple, and a reliable in vivo technique to assess VP as a surrogate measurement. Although a reliable technique, Miles assay is time-consuming, and the technique does not consider the compounding factors that may increase VP independently of endothelial-barrier regulation, such as blood pressure or blood flow. As an alternative, we describe fluorescein isothiocyanate-dextran lung permeability assay, a method that can also be adapted to measure VP and edema in other organs such as the brain and kidney.
虽然生理血管通透性 (VP) 介导了血浆、电解质、蛋白质和细胞穿过完整内皮屏障的选择性运输，但病理性 VP 导致内皮屏障完整性的丧失。虽然生理 VP 是正常宿主防御和组织修复的特征，但屏障功能受损可能导致异常的血管渗漏，同时发生组织水肿和炎症，最终导致危及生命的情况，如急性肺损伤或急性呼吸窘迫综合征、癌症、肾损伤等。VP 的测量有助于识别、设计和优化抗渗漏治疗。此外，它可以定义刺激或内皮屏障调节基因调控的效果。VP 的程度对于确定癌症的阶段和疾病预后可能很重要。本章讨论了 Miles 测定法，这是一种成熟、相对简单和可靠的体内技术，可作为替代测量方法来评估 VP。尽管 Miles 测定法是一种可靠的技术，但它很耗时，并且该技术不考虑可能独立于内皮屏障调节而增加 VP 的复合因素，例如血压或血流量。作为替代方法，我们描述了荧光素异硫氰酸酯-葡聚糖肺通透性测定法，该方法也可以适应测量大脑和肾脏等其他器官的 VP 和水肿。
