Brash James T, Ruhrberg Christiana, Fantin Alessandro
UCL Institute of Ophthalmology, University College London, London, UK.
Department of Biosciences, University of Milan, Milan, Italy.
Methods Mol Biol. 2022;2475:289-295. doi: 10.1007/978-1-0716-2217-9_21.
Before the endothelial mitogenic activity of the Vascular Endothelial Growth Factor A (VEGF) was described, VEGF had already been identified for its ability to induce vascular leakage. VEGF-induced vascular leakage has been most frequently studied in vivo using the Miles assay, a simple yet invaluable technique that has allowed researchers to unravel the molecular mechanisms underpinning vascular leakage both for VEGF and other permeability inducing agents. In this protocol, a mouse is intravenously injected with Evans Blue dye before VEGF is administered locally via intradermal injection. VEGF promotes vascular leak of serum proteins in the dermis, enabling Evans Blue-labeled albumin extravasation from the circulation and subsequent accumulation in the skin. As the volume of dye extravasation is proportional to the degree of vascular leak, it can be quantified as a proxy measurement of VEGF-induced vascular leakage.
在血管内皮生长因子A（VEGF）的内皮有丝分裂活性被描述之前，VEGF就因其诱导血管渗漏的能力而被发现。VEGF诱导的血管渗漏在体内最常使用迈尔斯试验进行研究，这是一种简单却极有价值的技术，它使研究人员能够揭示VEGF和其他通透性诱导剂导致血管渗漏的分子机制。在该实验方案中，在通过皮内注射局部给予VEGF之前，先给小鼠静脉注射伊文思蓝染料。VEGF促进真皮中血清蛋白的血管渗漏，使伊文思蓝标记的白蛋白从循环中渗出并随后在皮肤中积聚。由于染料渗出量与血管渗漏程度成正比，它可以作为VEGF诱导的血管渗漏的替代测量指标进行定量。
