, Volume 69, Issue 2, pp 229–244 | Cite as

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

  • Momoko Ishida
  • Kosuke Nishi
  • Kozue Shinohara
  • Nanami Kunihiro
  • Katsuhiro Osajima
  • Tomokazu Suemitsu
  • Takuya SugaharaEmail author
Original Article


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.


Immunostimulatory effect Dried bonito J774.1 cells Peritoneal macrophages Phagocytosis activity 



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.

Compliance with ethical standards

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.

Supplementary material

10616_2016_53_MOESM1_ESM.docx (227 kb)
Supplementary material 1 (DOCX 227 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Momoko Ishida
    • 1
  • Kosuke Nishi
    • 1
    • 2
  • Kozue Shinohara
    • 1
  • Nanami Kunihiro
    • 1
  • Katsuhiro Osajima
    • 3
  • Tomokazu Suemitsu
    • 3
  • Takuya Sugahara
    • 1
    • 2
    Email author
  1. 1.Faculty of AgricultureEhime UniversityMatsuyamaJapan
  2. 2.Food and Health Sciences Research CentreEhime UniversityMatsuyamaJapan
  3. 3.Senmi Ekisu Co., LtdOzuJapan

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