Determination of Tight Junction Integrity in Brain Endothelial Cells Based on Tight Junction Protein Expression.
Determination of Tight Junction Integrity in Brain Endothelial Cells Based on Tight Junction Protein Expression.
复制标题
基于紧密连接蛋白表达测定脑内皮细胞紧密连接完整性。
DOI：
10.1007/978-1-0716-3429-5_19
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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：
Alluri,Himakarnika;Peddaboina,ChanderSekhar;Tharakan,Binu
创伤性和缺血性损伤后发生的血脑屏障（BBB）功能障碍和通透性增高导致各种下游不良反应，如脑水肿和颅内压升高。由紧密连接蛋白组成的内皮细胞间紧密连接是BBB功能障碍和通透性增高的关键调节因子。BBB的主要紧密连接相关蛋白是闭合蛋白、密蛋白和细胞内连接至衔接蛋白闭合小带-1（ZO-1）的连接粘附分子。紧密连接相关蛋白质的定量测量提供了有价值的洞察屏障的完整性和机制，调节微血管通透性过高。蛋白质印迹分析是使用凝胶电泳分离和鉴定混合物中蛋白质的常用方法。了解这些蛋白质中的一种或多种的表达变化对于评估健康和疾病中的屏障完整性和渗透性至关重要。此外，研究它们将提供相关的下游信号通路的洞察力和评估的治疗方法调节血脑屏障通透性。在此，我们已经描述了免疫印迹分析的协议ZO-1作为一个指标，在脑微血管内皮细胞的紧密连接的完整性。
Blood-brain barrier (BBB) dysfunction and hyperpermeability that occurs following traumatic and ischemic insults lead to various downstream ill effects such as cerebral edema and elevation of intracranial pressure. The inter-endothelial tight junctions that consist of tight junction proteins are critical regulators of BBB dysfunctions and hyperpermeability. The major tight junction-associated proteins of the BBB are occludin, claudins, and junctional adhesion molecules that are intracellularly linked to the adaptor protein zonula occludens-1 (ZO-1). Quantitative measurement of tight junction-associated proteins provides valuable insight into barrier integrity and mechanisms that regulate microvascular hyperpermeability. Western blot analysis is a commonly used method to separate and identify proteins in a mixture using gel electrophoresis. Understanding the changes in the expression of one or more of these proteins is critical to evaluating barrier integrity and permeability in health and disease. Furthermore, studying them will provide insight into the associated downstream signaling pathways and evaluation of therapeutic approaches for regulating BBB permeability. Herein, we have described the protocol for immunoblot analysis of ZO-1 as an indicator of tight junction integrity in brain microvascular endothelial cells.