Siamwala Jamila, Swaminathan Akila, Chatterjee Suvro
Department of Molecular Pharmacology, Physiology and Biotechnology, Providence, RI, USA.
Warren Alpert Medical School of Brown University, Providence Veterans Affairs Medical Center, Providence, RI, USA.
Methods Mol Biol. 2024;2711:185-197. doi: 10.1007/978-1-0716-3429-5_15.
Vascular permeability, a tightly regulated process, is a direct measure of angiogenic and immune responses in the endothelium altered in several acute and chronic diseases such as sepsis, high-altitude pulmonary edema (HAPE), high-altitude cerebral edema (HACE), ischemia, and coronavirus disease 2019 (COVID-19) endotheliitis. Both endogenous and exogenous factors such as cytokines, chemokines, and hormones may affect vascular permeability. The conventional tools available for the measurement of vascular permeability in vitro and in vivo based on collagen-coated Transwell and dye-based spectrophotometric methods are indirect measures of permeability. In this chapter, we present our live in ovo protocols based on dextran-Texas red and avian chorioallantoic membrane assay developed using custom-made equipment to assess leakiness of endothelial cell barrier both in vitro and in vivo. Further, we validate this assay using different stressors such as ischemia and hypoxia known to affect endothelial barrier properties by potentiating actin stress fiber disorganization and disrupting the cell-cell junctions.
血管通透性是一个受到严格调控的过程，是衡量内皮细胞血管生成和免疫反应的直接指标，在多种急慢性疾病中会发生改变，如脓毒症、高原肺水肿（HAPE）、高原脑水肿（HACE）、局部缺血以及2019冠状病毒病（COVID-19）内皮炎症。细胞因子、趋化因子和激素等内源性和外源性因素均可影响血管通透性。基于胶原包被的Transwell和基于染料的分光光度法在体外和体内测量血管通透性的传统工具都是对通透性的间接测量。在本章中，我们介绍了基于葡聚糖-德州红和鸡胚绒毛尿囊膜试验的卵内实时实验方案，该方案使用定制设备开发，用于在体外和体内评估内皮细胞屏障的渗漏情况。此外，我们使用不同的应激源（如局部缺血和缺氧）验证了该试验，已知这些应激源会通过增强肌动蛋白应激纤维紊乱和破坏细胞间连接来影响内皮屏障特性。
