Measurement of Blood-Brain Barrier Hyperpermeability Using Evans Blue Extravasation Assay.
Measurement of Blood-Brain Barrier Hyperpermeability Using Evans Blue Extravasation Assay.
复制标题
使用伊文思蓝外渗测定法测量血脑屏障渗透性过高。
DOI：
10.1007/978-1-0716-3429-5_14
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Tharakan,Binu
中科院分区：
文献类型：
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作者：
Waithe,O'lisaYaa;Peng,Xu;Childs,EdW;Tharakan,Binu
血脑屏障（BBB）功能障碍和通透性过高与无数的脑病理学有关。伊文思蓝测定是研究脑疾病啮齿动物模型中BBB完整性和渗透性的最流行的方法之一。在正常生理条件下，血脑屏障对白蛋白是不可渗透的，因此静脉注射伊文思蓝时，伊文思蓝与血清白蛋白结合，并保持限制在血管内。在创伤性和缺血性损伤以及导致BBB高渗透性的其他脑病理中，邻近的内皮细胞部分地失去它们彼此的紧密接触，并且BBB变得对蛋白质（例如白蛋白）可渗透。伊文思蓝结合的白蛋白的这种细胞旁渗漏被认为是BBB功能障碍和通透性过高的可靠指标。在这里，我们描述了程序的BBB的完整性和高通透性的评价使用伊文思蓝外渗测定在小鼠模型的创伤性脑损伤。本文描述的方法侧重于静脉注射伊文思蓝，然后进行伊文思蓝染料提取。随后测量伊文思蓝的荧光强度，以确定染料外渗作为BBB通透性过高的直接指标。
Blood-brain barrier (BBB) dysfunction and hyperpermeability have been implicated in a myriad of brain pathologies. The Evans Blue assay is one of the most popular methods for studying BBB integrity and permeability in rodent models of brain disorders. Under normal physiological conditions, the BBB is impermeable to albumin, so Evans Blue when injected intravenously binds to serum albumin and remains restricted within blood vessels. In traumatic and ischemic injuries, and other brain pathologies that result in BBB hyperpermeability, neighboring endothelial cells partially lose their close contacts to each other, and the BBB becomes permeable to proteins such as albumin. This paracellular leak of Evans blue-bound albumin is considered a reliable indicator of BBB dysfunction and hyperpermeability. Here, we describe the procedures for the evaluation of BBB integrity and hyperpermeability using Evans Blue extravasation assay in a mouse model of traumatic brain injury. The method described here focuses on intravenous injection of Evans Blue followed by Evans Blue dye extraction. This is followed by the measurement of fluorescence intensity of Evans Blue to determine the dye extravasation as a direct indicator of BBB hyperpermeability.