Sodium caprate as an enhancer of macromolecule permeation across tricellular tight junctions of intestinal cells
Sodium caprate as an enhancer of macromolecule permeation across tricellular tight junctions of intestinal cells
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DOI：
10.1016/j.biomaterials.2012.09.051
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发表时间：
2013-01-01
期刊：
影响因子：
14
通讯作者：
Amasheh, Salah
中科院分区：
文献类型：
--
作者：
Krug, Susanne M.;Amasheh, Maren;Amasheh, Salah
癸酸钠是诱导药物吸收增强的有前途的候选者。迄今为止，这种增强吸收作用的机制尚未完全清楚。我们研究了癸酸在已建立的人类肠道细胞系 HT-29/B6 中如何作用于跨细胞（穿过细胞膜）和旁细胞（穿过紧密连接）途径的瞬时打开。癸酸钠 (10 mm) 引起跨上皮电阻快速且可逆的降低，通过双路阻抗谱测量，该电阻完全基于细胞旁通路的电阻变化。细胞旁标记通量的测量显示荧光素（330 Da）和 FITC-葡聚糖（4 和 10 kDa）的渗透性增加，表明细胞旁屏障打开。共聚焦显微镜显示三细胞紧密连接中的三纤维素蛋白和双细胞紧密连接中的claudin-5显着减少。这并不是由于蛋白质表达改变，因为 occludin、claudins 或 tricellulin 在蛋白质印迹中没有显着变化。细胞膜不可渗透的标记分子磺基-NHS-SS-生物素 (607 Da) 易位位点的可视化表明三细胞紧密连接是主要途径。我们认为癸酸盐对肠道药物摄取的已知增强作用是基于三细胞细胞接触的通透性增加，这是通过从三细胞紧密连接中可逆去除三纤维素来介导的。 (C) 2012 Elsevier Ltd. 保留所有权利。
Sodium caprate is a promising candidate for inducing drug absorption enhancement. The mechanism of that uptake-enhancing effect is not fully understood so far. We investigated how caprate acts in an established human intestinal cell line, HT-29/B6, on the transient opening of transcellular (across the cell membranes) and paracellular (across the tight junction) pathways. Sodium caprate (10 mm) caused a rapid and reversible decrease of transepithelial resistance which is based, as measured by two-path impedance spectroscopy, exclusively on resistance changes of the paracellular pathway. Measurements of paracellular marker fluxes revealed an increased permeability for fluorescein (330 Da) and FITC-dextran (4 and 10 kDa), indicating an opening of the paracellular barrier. Confocal microscopy revealed a marked reduction of tricellulin in tricellular tight junctions and of claudin-5 in bicellular tight junctions. This was not due to altered protein expression, as occludin, claudins or tricellulin were not significantly changed in Western blots. Visualization of the translocation site of the cell membrane-impermeable marker molecule sulpho-NHS-SS-biotin (607 Da) indicated the tricellular tight junction to be the predominant pathway. We suggest that caprate's known enhancing effect on intestinal drug uptake is based on increased permeability in tricellular cell contacts, mediated by reversible removal of tricellulin from the tricellular tight junction. (C) 2012 Elsevier Ltd. All rights reserved.