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Real‐time monitoring of trans‐epithelial electrical resistance in cultured intestinal epithelial cells: the barrier protection of water‐soluble dietary fiber

2017; Wiley; Volume: 18; Issue: 3 Linguagem: Inglês

10.1111/1751-2980.12456

1751-2980

Atsushi Majima, Osamu Handa, Yuji Naito, Yosuke Suyama, Yuriko Onozawa, Yasuki Higashimura, Katsura Mizushima, Mayuko Morita, Yukiko Uehara, Hideki Horie, Takaya Iida, Akifumu Fukui, Osamu Dohi, Tetsuya Okayama, Naohisa Yoshida, Kazuhiro Kamada, Kazuhiro Katada, Kazuhiko Uchiyama, Takeshi Ishikawa, Tomohisa Takagi, Hideyuki Konishi, Zenta Yasukawa, Makoto Tokunaga, Tsutomu Ōkubo, Yoshito Itoh,

Connexins and lens biology

OBJECTIVES In this study we aimed to verify a real‐time trans‐epithelial electrical resistance ( TEER) monitoring system in a C aco‐2 monolayer and to investigate the therapeutic effect of partially hydrolyzed guar gum ( PHGG ), a dietary fiber, against interferon ( IFN )‐γ‐induced intestinal barrier dysfunction using this monitoring system. METHODS We measured TEER using a real‐time monitoring system and evaluated epithelial paracellular permeability using fluorescein isothiocyanate‐conjugated dextran (4 kDa; FD 4) in C aco‐2 monolayers treated with IFN ‐γ for 48 h. The expression and distribution of tight junction ( TJ )‐associated proteins, ZO ‐1 and occludin, were analyzed by W estern blot and immunocytochemistry, respectively. In some experiments PHGG was added prior to IFN ‐γ treatment in order to investigate its protective effect on barrier function. RESULTS IFN ‐γ treatment significantly decreased TEER and increased FD 4 flux across C aco‐2 monolayers, indicating a great influence of IFN ‐γ on the intestinal epithelial paracellular permeability. In contrast, the pretreatment of PHGG significantly reduced the IFN ‐γ‐induced increment of FD 4 flux without affecting TEER . Neither IFN ‐γ nor PHGG treatment affected the expressions of TJ ‐associated proteins, while immunocytochemistry showed that IFN ‐γ‐induced redistribution of occludin was clearly restored by PHGG . CONCLUSIONS Real‐time TEER monitoring enabled us to evaluate the dynamic changes of intestinal epithelial barrier function. PHGG may have a protective effect against IFN ‐γ‐induced barrier dysfunction by attenuating the paracellular hyperpermeability; thus, its promotion as a functional food is anticipated.