Poly-L-arginine-induced internalization of tight junction proteins increases the paracellular permeability of the Caco-2 cell monolayer to hydrophilic macromolecules
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概要
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We investigated whether poly-L-arginine (PLA) enhances the paracellular permeability of the Caco-2 monolayer to hydrophilic macromolecules and clarified the disposition of tight junction (TJ) proteins. The transepithelial electrical resistance (TEER) and FITC-dextran (FD-4) permeation were determined after treatment with PLA. TJ proteins were visualized using immunofluorescence microscopy after PLA exposure and depletion, and their expression levels were determined. The barrier function of TJs was also evaluated by measuring the alterations in the TEER and in the localization of TJ proteins. PLA induced an increase in hydrophilic macromolecule, FD-4, permeation through Caco-2 cell monolayers and a decrease in the TEER in a concentration-dependent manner, without any significant impact on the cell viability. This increased paracellular permeability induced by PLA was found to be internalized of claudin-4, ZO-1, tricellulin and mainly occludin from cell-cell junction to the subcellular space. ZO-1 appeared to play an important role in the reconstitution of TJ strand structures following PLA depletion. These results indicate that the PLA led to the internalization of TJ proteins to the subcellular space, subsequently increasing the permeability of the Caco-2 cell monolayer to FD-4 via a paracellular route.
著者
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Juni Kazuhiko
Faculty of Pharmaceutical Sciences, Josai University
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Uchida Masaki
Faculty Of Pharmaceutical Sciences Josai University
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Juni Kazuhiko
Faculty Of Pharmaceutical Sciences Hokkaido University
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Morimoto Yasunori
Faculty Of Pharmaceutical Sciences Josai University
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Uchida Hiroyuki
Faculty Of Engineering Fukui University
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Natsume Hideshi
Faculty Of Pharmaceutical Sciences Josai University:research Institute Of Tts Technology
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Ohtake Kazuo
Faculty of Pharmaceutical Sciences, Josai University
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Kobayashi Jun
Faculty Of Agriculture Yamaguchi University
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Yamaki Tsutomu
Faculty of Pharmaceutical Sciences, Josai University
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Ichikawa Keiko
Faculty of Pharmaceutical Sciences, Josai University
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Ohshima Shinji
Faculty of Pharmaceutical Sciences, Josai University
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