Aguanno Doriane, Postal Bárbara Graziela, Carrière Véronique, Thenet Sophie
Centre de Recherche Saint-Antoine, INSERM UMRS 938, Sorbonne Université, Paris, France.
EPHE, PSL University, Paris, France.
Methods Mol Biol. 2021;2367:1-11. doi: 10.1007/7651_2021_367.
An increased intestinal permeability has been described in many diseases including inflammatory bowel disease and metabolic disorders, and a better understanding of the contribution of intestinal barrier impairment to pathogenesis is needed. In recent years, attention has been paid to the leak pathway, which is the route of paracellular transport allowing the diffusion of macromolecules through the tight junctions of the intestinal epithelial lining. While the passage of macromolecules by this pathway is very restricted under physiological conditions, its amplification is thought to promote an excessive immune activation in the intestinal mucosa. The Ussing chambers have been widely used to measure both active and passive transepithelial fluxes in intact tissues. In this chapter we present how this simple device can be used to measure paracellular permeability to macromolecules in the mouse intestine. We propose a detailed protocol and describe how to best exploit all the possibilities of this technique, correctly interpret the results, and avoid the main pitfalls.
在包括炎症性肠病和代谢紊乱在内的许多疾病中，肠道通透性均有所增加，因此需要更好地了解肠道屏障损伤在发病机制中的作用。近年来，人们开始关注渗漏途径，这是一种细胞旁转运途径，允许大分子通过肠上皮内衬的紧密连接进行扩散。虽然在生理条件下，大分子通过该途径的转运非常有限，但人们认为其增强会促进肠黏膜中过度的免疫激活。尤斯灌流小室已被广泛用于测量完整组织中的主动和被动跨上皮通量。在本章中，我们将介绍如何使用这种简单的装置来测量小鼠肠道对大分子的细胞旁通透性。我们提出了一个详细的方案，并描述了如何充分利用该技术的所有可能性、正确解释结果以及避免主要陷阱。
