Evaluating Vascular Hyperpermeability-inducing Agents in the Skin with the Miles Assay.
Evaluating Vascular Hyperpermeability-inducing Agents in the Skin with the Miles Assay.
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DOI：
10.3791/57524
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发表时间：
2018-06-19
期刊：
影响因子：
--
通讯作者：
Fantin A
中科院分区：
文献类型：
--
作者：
Brash JT;Ruhrberg C;Fantin A
脊椎动物生物体中血管内皮的主要功能是充当血液和身体的每个组织之间的屏障，由此内皮对血细胞、血浆大分子和水的渗透性可以根据生理需要进行调整。在某些疾病中，释放靶向内皮屏障的细胞因子和生长因子以瞬时增加血管通透性;然而，它们的长期存在可能导致慢性血管通透性过高，从而导致组织损伤性水肿。Miles试验是一种体内技术，允许研究人员通过血管渗漏的替代测量来研究血管通透性过高。在这里，我们提供了一个详细的协议，如何在小鼠，这是最广泛使用的模式生物，研究哺乳动物的生理和病理学进行这一程序。该程序涉及静脉内注射伊文思蓝染料以标记循环白蛋白，然后向小鼠的相对侧腹多次皮内注射渗透诱导剂和溶媒对照溶液。因此，伊文思蓝染料逐渐渗漏到真皮中，在真皮中其积累并可被提取用于定量，作为相对于媒介物由渗透诱导剂诱导的渗漏。Miles测定可以在野生型或遗传修饰的小鼠模型中进行，并且可以与药物施用组合以研究调节血管通透性的分子机制并鉴定能够诱导或阻断高通透性的试剂/靶标。
The primary function of the vascular endothelium in vertebrate organisms is to serve as a barrier between the blood and each tissue of the body, whereby the permeability of the endothelium to blood cells, plasma macromolecules, and water can be adapted according to the physiological need. In certain diseases, cytokines and growth factors are released that target the endothelial barrier to transiently increase vascular permeability; however, their prolonged presence may cause chronic vascular hyperpermeability and thereby tissue-damaging edema. The Miles assay is an in vivo technique that allows researchers to study vascular hyperpermeability through the proxy measurement of vascular leakage. Here, we provide a detailed protocol on how to perform this procedure in the mouse, which is the most widely used model organism to study mammalian physiology and pathology. The procedure involves the intravenous injection of Evans blue dye to label the circulating albumin followed by multiple intradermal injections of permeability-inducing agents and vehicle control solutions into opposing flanks of the mouse. Consequently, Evans blue dye gradually leaks into the dermis, where it accumulates and can be extracted for quantification as leakage induced by the permeability-inducing agent relative to the vehicle. The Miles assay can be performed in wild type or genetically modified mouse models and may be combined with drug administration to study molecular mechanisms that regulate vascular permeability and identify agents/targets capable of inducing or blocking hyperpermeability.
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影响因子：
9.8
作者：
Nagy, Janice A.;Benjamin, Laura;Zeng, Huiyan;Dvorak, Ann M.;Dvorak, Harold F.
通讯作者：
Dvorak, Harold F.
DOI：
10.1084/jem.20111343
发表时间：
2012-07-02
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Sun Z;Li X;Massena S;Kutschera S;Padhan N;Gualandi L;Sundvold-Gjerstad V;Gustafsson K;Choy WW;Zang G;Quach M;Jansson L;Phillipson M;Abid MR;Spurkland A;Claesson-Welsh L
通讯作者：
Claesson-Welsh L
影响因子：
20.3
作者：
Acevedo, Lisette M.;Barillas, Samuel;Cheresh, David A.
通讯作者：
Cheresh, David A.
影响因子：
7.3
作者：
Roth, Lise;Prahst, Claudia;Augustin, Hellmut G.
通讯作者：
Augustin, Hellmut G.
影响因子：
3.3
作者：
Strbian, D.;Durukan, A.;Tatlisumak, T.
通讯作者：
Tatlisumak, T.