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The three isoforms of hepcidin in human serum and their processing determined by liquid chromatography-tandem mass spectrometry (LC-tandem MS)

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Abstract

Hepcidin, the iron regulatory hormone, has three isoforms; -20, -22 and -25. While hepcidin-25 has been studied extensively, the physiological significance of other isoforms remains poorly understood. Using a quantitative method based on liquid chromatography-tandem mass spectrometry (LC-tandem MS) developed by our group, we quantified hepcidin isoforms in human serum to elucidate their characteristics, and investigated the role of hepatocytes in isoform processing. Hepcidin isoforms in serum obtained from 40 healthy volunteers were quantified. Synthetic hepcidin peptides were added to healthy serum, and to HepG2 culture media, and hepcidin isoform concentrations determined. All three hepcidin isoforms were detected in human serum; however, hepcidin-25 concentrations were highest. The three hepcidin isoforms showed a strong positive correlation with each other and with serum ferritin. Additionally, while hepcidin-20 was strongly correlated with serum creatinine, the other isoforms were not. Hepcidin-20 and -25 levels were also increased in chronic kidney disease (CKD) serum. Hepcidin-22 rapidly degraded into hepcidin-20, whereas hepcidin-25 remained relatively stable. Finally, hepcidin-22 degradation into hepcidin-20 was accelerated in the presence of HepG2. This method has enabled us to reveal fundamental characteristics of the three hepcidin isoforms in serum and may be a powerful tool for quantifying hepcidin isoform expression and processing.

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Author information

Authors and Affiliations

  1. Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa, Hokkaido, 078-8510, Japan

    Lynda Addo, Katsuya Ikuta, Yasumichi Toki, Mayumi Hatayama, Masayo Yamamoto, Satoshi Ito, Motohiro Shindo, Mikihiro Fujiya & Yutaka Kohgo

  2. Department of Gastrointestinal Immunology and Regenerative Medicine, Asahikawa Medical University, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido, 078-8510, Japan

    Hiroki Tanaka & Yutaka Kohgo

  3. Product Research Department, Kamakura Research Labs, Chugai Pharmaceutical Co., Ltd., 200 Kajiwara, Kamakura, Kanagawa, 247-8530, Japan

    Yusuke Sasaki & Yasushi Shimonaka

Authors
  1. Lynda Addo
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  2. Katsuya Ikuta
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  3. Hiroki Tanaka
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  4. Yasumichi Toki
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  5. Mayumi Hatayama
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  6. Masayo Yamamoto
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  7. Satoshi Ito
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  8. Motohiro Shindo
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  9. Yusuke Sasaki
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  10. Yasushi Shimonaka
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  11. Mikihiro Fujiya
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Corresponding author

Correspondence to Katsuya Ikuta.

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Conflict of interest

Lynda Addo, Katsuya Ikuta, Yasumichi Toki, Mayumi Hatayama, Masayo Yamamoto, Satoshi Ito, Motohiro Shindo, Mikihiro Fujiya, Yutaka Kohgo (Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical University) and Hiroki Tanaka (Department of Gastrointestinal Immunology and Regenerative Medicine, Asahikawa Medical University) received research funding from Chugai Pharmaceutical Co. Ltd. This study was also performed in collaboration with Chugai Pharmaceutical Co. Ltd. These departments also received collaborative research funding for research work concerning iron metabolism from Novartis Pharma K. K. and Asahi Kasei Medical Co. Ltd.

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Addo, L., Ikuta, K., Tanaka, H. et al. The three isoforms of hepcidin in human serum and their processing determined by liquid chromatography-tandem mass spectrometry (LC-tandem MS). Int J Hematol 103, 34–43 (2016). https://doi.org/10.1007/s12185-015-1885-y

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  • DOI: https://doi.org/10.1007/s12185-015-1885-y

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