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Non-targeted metabolite profiling in activated macrophage secretion

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Abstract

Periodontal diseases are inflammatory infectious diseases that affect the periodontal tissue. Macrophages play a central role in inflammatory conditions, leading to the destruction of tissues. Identifying the signaling molecules secreted by macrophages would be valuable to the study of these diseases. Here, we present non-targeted analysis using capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) for the profiling of extracellular metabolites released during macrophage activation. Lipopolysaccharide (LPS)-induced activation of a mouse macrophage-like cell line RAW264.7 was used as a model system. Cells were treated without (control) or with LPS for 22 h and, after washing, were incubated for 1 h in phosphate-buffered saline. The accumulation of metabolites in the culture supernatant was monitored. LPS treatment significantly enhanced the accumulation of prostaglandins, tumor necrosis factor-α, nitric oxide and citrulline in the culture medium. RAW264.7 cells produced 46 metabolites and 66% of these showed significant changes (P < 0.05) following cell activation. In particular, the production of leucine, hypoxanthine, choline, putrecine, N 8-acetylspermidine, succinate, itaconate, and 4-methyl-2-oxopentanoate was significantly increased by cell activation (P < 0.001). Significantly elevated production of lactate and glycine was also observed. Here, we present the first catalog of the up and down-regulation of the various metabolites secreted by macrophages following inflammatory activation.

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Acknowledgments

This work was supported by research funds from the Yamagata Prefectural Government and the city of Tsuruoka. We thank Shinobu Abe for technical assistance.

Conflicts of interest

The authors have no conflicts of interest to declare.

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Authors and Affiliations

  1. Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata, 997-0017, Japan

    Masahiro Sugimoto, Hiromi Onuma, Miku Kaneko, Masayo Mori, Yasuko Sakaguchi, Tomoyoshi Soga & Masaru Tomita

  2. Systems Biology Program, Graduate School of Media and Governance, Keio University, Fujisawa, Kanagawa, 252-8520, Japan

    Masahiro Sugimoto, Tomoyoshi Soga & Masaru Tomita

  3. Divisions of Pharmacology, Meikai University School of Dentistry, Sakado, Saitama, 350-0283, Japan

    Hiroshi Sakagami

  4. Faculty of Science, Josai University, Sakado, Saitama, 350-0295, Japan

    Yoshiko Yokote

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  1. Masahiro Sugimoto
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  2. Hiroshi Sakagami
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  3. Yoshiko Yokote
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  4. Hiromi Onuma
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  8. Tomoyoshi Soga
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  9. Masaru Tomita
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Correspondence to Masahiro Sugimoto.

Additional information

M. Sugimoto and H. Sakagami contributed equally to this work.

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Sugimoto, M., Sakagami, H., Yokote, Y. et al. Non-targeted metabolite profiling in activated macrophage secretion. Metabolomics 8, 624–633 (2012). https://doi.org/10.1007/s11306-011-0353-9

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  • DOI: https://doi.org/10.1007/s11306-011-0353-9

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