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Interleukin 10 inhibits interferon γ- and tumor necrosis factor α-stimulated activation of NADPH oxidase 1 in human colonic epithelial cells and the mouse colon

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Abstract

Background

NADPH oxidase 1 (Nox1) is preferentially expressed in the colon, but its functional role is not fully understood. This study was designed to elucidate a potential role of Nox1 in inflammation of the colon.

Methods

Superoxide production by T84 cells was measured by the cytochrome c method. Protein and mRNA levels of Nox1 and Nox organizer 1 (NOXO1) in the cells were measured by real-time reverse transcriptase PCR and Western blotting, respectively. Expression of Nox1, Nox2, dual oxidase 2 (Duox2), NOXO1, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α mRNAs was measured in proximal, middle, and distal portions of colonic mucosas from male wild-type C57BL/6J and interleukin (IL)-10 knockout mice at 6, 10, and 16 weeks of age. Grading of inflammation was done by scoring histological changes.

Results

IL-10 significantly inhibited IFN-γ- or TNF-α-induced up-regulation of superoxide-producing activity in T84 cells by suppressing expression of Nox1 mRNA and protein. IL-10 also inhibited TNF-α-stimulated induction of NOXO1 and p38 MAPK phosphorylation. Levels of Nox1, but not Nox2 or Duox2 mRNA, was age-dependently increased following a gradient with low levels in the proximal colon and high levels in the distal colon of the wild-type mice. The absence of IL-10 significantly facilitated Nox1 expression in association with increased IFN-γ mRNA expression before the development of spontaneous colitis and age-dependently accelerated their mRNA expression.

Conclusions

IL-10 may be a possible down-regulator of the Nox1-based oxidase in the colon, suggesting a potential role of reactive oxygen species (ROS) derived from Nox1-based oxidase in inflammation of the colon.

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Authors and Affiliations

  1. Department of Stress Science, Institute of Health Biosciences, The University of Tokushima Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan

    Mai Kamizato, Kensei Nishida, Kiyoshi Masuda, Keiko Takeo, Yuta Yamamoto, Tomoko Kawai, Shigetada Teshima-Kondo, Toshihito Tanahashi & Kazuhito Rokutan

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  1. Mai Kamizato
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  2. Kensei Nishida
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Correspondence to Kensei Nishida.

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Kamizato, M., Nishida, K., Masuda, K. et al. Interleukin 10 inhibits interferon γ- and tumor necrosis factor α-stimulated activation of NADPH oxidase 1 in human colonic epithelial cells and the mouse colon. J Gastroenterol 44, 1172–1184 (2009). https://doi.org/10.1007/s00535-009-0119-6

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  • DOI: https://doi.org/10.1007/s00535-009-0119-6

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