Department of Gastroenterology, Infectious Diseases and Rheumatology, Campus Benjamin Franklin, Charité Berlin, Berlin, Germany.
Gut. 2012 Feb;61(2):220-8. doi: 10.1136/gutjnl-2011-300123. Epub 2011 Aug 24.
Epithelial barrier defects are well known in coeliac disease, but the mechanisms are only poorly defined. It is unclear, whether barrier disturbance reflects upregulated epithelial transcytosis or paracellular leakage.
To characterise the molecular structure and function of the epithelial tight junction (TJ) and mechanisms of its dysregulation.
Molecular analysis of proteins involved in TJ assembly and their regulation was performed by western blotting and confocal microscopy correlated to electrophysiology.
A complex alteration of the composition of epithelial TJ proteins (with more pore-forming claudins like claudin-2 and a reduction in tightening claudins like claudin-3, -5 and -7) was found for protein expression and subcellular localisation, responsible for an increase in paracellular biotin-NHS uptake. In contrast, epithelial apoptosis was only moderately elevated (accounting for a minor portion of barrier defects) and epithelial gross lesions--for example, at cell extrusion zones, were absent. This TJ alteration was linked to an altered localisation/expression of proteins regulating TJ assembly, the polarity complex protein Par-3 and the serine-/threonine phosphatase PP-1.
Changes in cell polarity proteins Par-3 and PP-1 are associated with altered expression and assembly of TJ proteins claudin-2, -3, -5 and -7 and ZO-1, causing paracellular leakage in active coeliac disease.
肠病相关的细胞屏障缺陷是众所周知的,但相关机制仍不完全明确。目前尚不清楚这种屏障障碍是反映了上皮细胞的转胞吞作用增强,还是细胞旁渗漏增加。
描述上皮细胞紧密连接(TJ)的分子结构和功能及其调节机制。
通过 Western blot 和共聚焦显微镜与电生理学相关联,对参与 TJ 组装的蛋白质及其调节机制的分子进行分析。
发现上皮 TJ 蛋白的组成发生了复杂的改变(更多的孔形成闭合蛋白,如闭合蛋白-2,而收紧闭合蛋白,如闭合蛋白-3、-5 和-7 的减少),这导致了细胞旁生物素-NHS 摄取的增加。相比之下,上皮细胞凋亡仅适度增加(占屏障缺陷的一小部分),并且不存在上皮细胞的大病变,例如细胞挤出区。这种 TJ 改变与调节 TJ 组装的蛋白质、极性复合物蛋白 Par-3 和丝氨酸/苏氨酸磷酸酶 PP-1 的定位/表达改变有关。
细胞极性蛋白 Par-3 和 PP-1 的变化与 TJ 蛋白闭合蛋白-2、-3、-5 和-7 和 ZO-1 的表达和组装改变有关,导致活动性乳糜泻中的细胞旁渗漏。