Ramnath Raina D, Satchell Simon C
Bristol Renal, Translational Health Sciences, Bristol Medical School, University of Bristol, Bristol, UK.
Methods Mol Biol. 2020;2067:145-151. doi: 10.1007/978-1-4939-9841-8_11.
Endothelial cells form the inner lining of all blood vessels and play a vital role in regulating vascular permeability. This applies to the circulation in general and also to specific capillary beds including the renal glomerular capillaries. Endothelial dysfunction, including increased permeability, is a key component of diabetes-induced organ damage. Endothelial cells together with their glycocalyx, grown on porous membranes, provide an excellent model to study endothelial permeability properties. Here we describe the measurement of two characteristics of glomerular endothelial cell (GEnC) monolayers: electrical resistance and macromolecular passage. Trans-endothelial electrical resistance provides a measure of small-pore pathways across the endothelium and provides an index of monolayer confluence and cell-cell junction integrity. Measurement of macromolecular passage provides an index of large-pore pathways and use of labeled albumin provides direct relevance to the clinically important parameter of albuminuria. The combination of the two approaches provides a fantastic tool to elucidate endothelial barrier function in vitro including in response to cytokines, pathological stimuli, and potential therapeutic agents.
内皮细胞构成所有血管的内层,在调节血管通透性方面发挥着至关重要的作用。这适用于一般的循环系统,也适用于包括肾肾小球毛细血管在内的特定毛细血管床。内皮功能障碍,包括通透性增加,是糖尿病诱导的器官损伤的关键组成部分。生长在多孔膜上的内皮细胞及其糖萼提供了一个研究内皮通透性特性的优秀模型。在这里,我们描述了肾小球内皮细胞(GEnC)单层的两个特性的测量:电阻和大分子通过。跨内皮电阻提供了一种测量穿过内皮的小孔径通路的方法,并提供了单层汇合度和细胞间连接完整性的指标。大分子通过的测量提供了大孔径通路的指标,使用标记白蛋白与蛋白尿这一临床重要参数直接相关。这两种方法的结合提供了一个极好的工具,用于阐明体外内皮屏障功能,包括对细胞因子、病理刺激和潜在治疗药物的反应。