Utech Markus, Brüwer Matthias, Nusrat Asma
Department of Pathology and Laboratory Medicine, Epithelial Pathobiology Unit, Emory University School of Medicine, Atlanta, GA, USA.
Methods Mol Biol. 2006;341:185-95. doi: 10.1385/1-59745-113-4:185.
Chronic inflammation in mucosal tissues can influence epithelial barrier function via pro-inflammatory cytokines such as interferon (IFN)-gamma and tumor necrosis factor-alpha. Increased mucosal levels of these cytokines have been observed in mucosal biopsies from patients with a chronic inflammatory condition referred to as inflammatory bowel disease. Paracellular permeability across epithelial cells is regulated by tight junctions (TJs), which are the apical most junctions in epithelial cells. Given that pro-inflammatory cytokines modulate the epithelial barrier and that TJs regulate epithelial permeability, we analyzed the influence of IFN-gamma on U function/structure. Our results suggest that IFN-gamma induced a time-dependent increase in paracellular permeability that was associated with internalization of TJ transmembrane proteins, occludin, junction adhesion molecule A, and claudin-1. In this chapter, we focus on selected methods used to investigate the influence of IFN-gamma on epithelial barrier function.
黏膜组织中的慢性炎症可通过促炎细胞因子，如干扰素（IFN）-γ和肿瘤坏死因子-α，影响上皮屏障功能。在一种称为炎症性肠病的慢性炎症性疾病患者的黏膜活检中，已观察到这些细胞因子在黏膜中的水平升高。上皮细胞间的旁细胞通透性由紧密连接（TJ）调节，紧密连接是上皮细胞最顶端的连接。鉴于促炎细胞因子调节上皮屏障，且紧密连接调节上皮通透性，我们分析了IFN-γ对紧密连接功能/结构的影响。我们的结果表明，IFN-γ诱导旁细胞通透性随时间增加，这与紧密连接跨膜蛋白、闭合蛋白、连接黏附分子A和Claudin-1的内化有关。在本章中，我们重点介绍用于研究IFN-γ对上皮屏障功能影响的选定方法。
