Difructose anhydride III and sodium caprate activate paracellular transport via different intracellular events in Caco-2 cells
Difructose anhydride III and sodium caprate activate paracellular transport via different intracellular events in Caco-2 cells
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DOI：
10.1016/j.lfs.2006.01.044
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发表时间：
2006-06-20
期刊：
影响因子：
6.1
通讯作者：
Hara, Hiroshi
中科院分区：
文献类型：
--
作者：
Suzuki, Takuya;Hara, Hiroshi
DFA III是一种不可消化的二糖，可通过紧密连接(TJ)激活钙转运，但DFA III促进细胞旁转运的特性和机制尚不清楚。我们利用Caco-2单层比较了DFAIII和已知的TJ通透性增强剂--CAPRATE(C10)对TJ蛋白的变化、细胞旁标志物的转运以及9种细胞信号阻滞剂的作用。在Caco-2单层的顶端培养液中加入DFAIII(0-100mmoL/L)和C10(0-10mmoL/L)呈剂量依赖性地降低跨上皮电阻(TER)，这是TJ通透性的一个指标。C10的还原速度远快于DFAIII。加入100mmoL/L DFAIII和10mmoL/L C10可促进不同分子量(182-43,200)的胞外标志物的转运。在C10存在下的转运速率比DFAIII大得多，而两种处理对TER的减少相似(从1000欧加厘米(2)减少到300欧加厘米(2))。DFAIII和C10处理改变了肌动蛋白细丝和Claudin-1的分布，但不改变阻滞素、连接黏附分子-1或闭锁带-1的分布；然而，TJ蛋白模式的变化因处理而不同。肌球蛋白轻链激酶的抑制剂和细胞内钙离子的螯合剂([Ca~(2+)](I))可减弱C10对TER的还原，但不能减弱DFAIII的作用。这些数据表明，DFAIII诱导的TJ通透性增加是通过[Ca~(2+)](I)非依赖机制改变肌动蛋白和claudin-1的结果。(C)2006 Elsevier Inc.保留所有权利。
A nondigestible disaccharide, difiructose anhydride (DFA) III, is known to activate calcium transport via tight junctions (TJs); however, the characteristics of and mechanisms for the increase in paracellular transport induced by DFAIII have not been clarified. We compared the effect of DFAIII with that of sodium caprate (C10), a well-known enhancer of TJ permeability, on the changes in TJ proteins, transport of paracellular markers, and effects of nine cellular signaling blockers using Caco-2 monolayers. The addition of DFAIII (0-100 mmol/L) and C10 (0-10 mmol/L) to the apical medium of the Caco-2 monolayers dose-dependently decreased transepithelial electrical resistance (TER), which is an indicator of TJ permeability. The reduction with C10 was much faster than that with DFAIII. Transport of the paracellular markers of various molecular weights (182-43,200) was elevated by the addition of 100 mmol/L DFAIII and 10 mmol/L C10. The transport rates were much in the presence of C10 than of DFAIII, while the reduction in TER by two treatments was similar (from 1000 to 300 Omega cm(2)). Treatment with DFAIII and C10 changed the distribution of actin filament and claudin-1, but not occludin, junctional adhesion molecule-1, or zonula occludens-1; however, alterations in the patterns of the TJ proteins differed according to treatment. An inhibitor of myosin light chain kinase and a chelator of intracellular calcium ion ([Ca2+](i)) attenuated the TER reduction by C10, but not by DFAIII. These data demonstrate that the increase in TJ permeability induced by DFAIII results from the alterations to actin and claudin-1 via [Ca2+](i)-independent mechanisms. (c) 2006 Elsevier Inc. All rights reserved.