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Immunostimulatory effect of dried bonito extract on mouse macrophage cell lines and mouse primary peritoneal macrophages

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Abstract

Dried bonito is a preserved food used in Japan, which contains abundant flavor ingredients and functional substances. We focused on the immunostimulatory effect of dried bonito extract (DBE) on mouse macrophage-like J774.1 cells, RAW264.7 cells, and mouse primary peritoneal macrophages. DBE significantly stimulated the production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) by both J774.1 cells and peritoneal macrophages by enhancing the cytokine gene expression levels. In addition, DBE stimulated nitric oxide production by enhancing the expression of inducible nitric oxide synthase in RAW264.7 cells. DBE also increased the phagocytosis activity of J774.1 cells. Immunoblot analysis revealed that DBE has an immunostimulatory effect on macrophages through activation of mitogen-activated protein kinase and nuclear factor-κB cascades. TNF-α production enhanced by DBE was partially inhibited by treatment with TLR4 inhibitor TAK-242, whereas IL-6 production enhanced by DBE was almost inhibited. These results suggested that DBE is thought to strongly stimulate the TLR4 signaling pathway for macrophage activation, and its activation is also involved in other signaling. Finally, the phagocytosis activity of peritoneal macrophages from DBE-administered BALB/c mice increased significantly, suggesting that DBE has the potential to stimulate macrophage activity in vivo. In conclusion, these data indicated that DBE contributes to activating host defense against pathogens by activating innate immunity.

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Acknowledgements

We thank members of the Advanced Research Support Center (ADRES), Ehime University for the use of animal facility.

Author contributions

MI, KN, KS, NK, and TS conceived and designed the experiments. MI and KS, and NK performed the experiments and analyzed the data. MI, KN, and TS wrote the manuscript.

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Authors and Affiliations

  1. Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime, 790-8566, Japan

    Momoko Ishida, Kosuke Nishi, Kozue Shinohara, Nanami Kunihiro & Takuya Sugahara

  2. Food and Health Sciences Research Centre, Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime, 790-8566, Japan

    Kosuke Nishi & Takuya Sugahara

  3. Senmi Ekisu Co., Ltd, 779-2 Noda, Hirano-cho, Ozu, Ehime, 795-0021, Japan

    Katsuhiro Osajima & Tomokazu Suemitsu

Authors
  1. Momoko Ishida
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  2. Kosuke Nishi
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  3. Kozue Shinohara
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  4. Nanami Kunihiro
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  5. Katsuhiro Osajima
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  6. Tomokazu Suemitsu
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  7. Takuya Sugahara
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Corresponding author

Correspondence to Takuya Sugahara.

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Conflict of interest

Katsuhiro Osajima, Tomokazu Suemitsu, and Takuya Sugahara are inventors in a patent application filed by Ehime University, which disclose bioactive agents targeting immunostimulation described in the present article. The remaining authors declare no conflict of interest.

Additional information

Practical Application: Although dried bonito contains abundant flavor ingredients and functional substances, it was unclear whether dried bonito affects macrophage activation leading to enhancement of the immune system. In this study, the extract from dried bonito showed immunostimulatory effects on experimental animals as well as on cell lines. Our findings suggest that DBE would be a novel functional food for immunostimulation with academic significance and that daily intake of DBE contributes to the maintenance of human health with enhancement of the immunity due to macrophage activation.

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Ishida, M., Nishi, K., Shinohara, K. et al. Immunostimulatory effect of dried bonito extract on mouse macrophage cell lines and mouse primary peritoneal macrophages. Cytotechnology 69, 229–244 (2017). https://doi.org/10.1007/s10616-016-0053-4

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  • DOI: https://doi.org/10.1007/s10616-016-0053-4

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