Molecular and Cellular Biochemistry

, Volume 452, Issue 1–2, pp 51–62 | Cite as

High-glucose-induced changes in macrophage secretome: regulation of immune response

  • Taniya Kaewarpai
  • Visith ThongboonkerdEmail author


Secretory products from infiltrating macrophages have been thought to play crucial roles in development and progression of diabetic complications in various tissues/organs. Nevertheless, diabetes-induced changes in macrophage secretory products remained largely unknown. We thus analyzed high-glucose (HG)-induced changes in secretome of human macrophages derived from U937 human monocytic cell line after phorbol 12-myristate 13-acetate (PMA) activation. Serum-free culture supernatants were collected from macrophages exposed to 5.5 mM glucose (NG-M-sup) (normal control), 25 mM glucose (HG-M-sup), or 5.5 mM glucose + 19.5 mM mannitol (MN-M-sup) (osmotic control) for 16 h. After dialysis and lyophilization, secreted proteins were subjected to 2-DE analysis (n = 5 gels derived from 5 independent cultures per group). Quantitative analysis and statistics revealed 23 protein spots whose secretory levels significantly differed among the three conditions. These proteins were successfully identified by nanoLC-ESI-MS/MS analyses and changes in levels of heat shock protein 90 (HSP90), HSP70, HSP60, and β-actin were confirmed by Western blotting. Global protein network and functional enrichment analyses revealed that the altered proteins in HG-M-sup were involved mainly in regulation of immune response that might communicate with other bystander cells through the release of extracellular vesicles. These data may lead to a wider view of pathogenic mechanisms of diabetic complications.


Diabetes Diabetic complications Mass spectrometry Proteome Proteomic analysis Secretory proteins 



We are grateful to Dr. Phornpimon Tipthara for her technical assistance. This study was supported by Mahidol University research grant and the Thailand Research Fund (IRN60W0004 and IRG5980006). TK was also supported by Siriraj Graduate Thesis Scholarship.

Author Contributions

TK and VT designed research; TK performed experiments; TK and VT analyzed data; TK and VT wrote the manuscript; All authors reviewed and approved the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj HospitalMahidol UniversityBangkokThailand
  2. 2.Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj HospitalMahidol UniversityBangkokThailand
  3. 3.Center for Research in Complex Systems ScienceMahidol UniversityBangkokThailand
  4. 4.Medical Proteomics Unit, Office for Research and Development, Siriraj HospitalMahidol UniversityBangkokThailand

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