Molecular and Cellular Biochemistry

, Volume 461, Issue 1–2, pp 1–14 | Cite as

Proteomic analysis of peripheral blood polymorphonuclear cells (PBMCs) reveals alteration of neutrophil extracellular trap (NET) components in uncontrolled diabetes

  • J. SoongsathitanonEmail author
  • W. Umsa-Ard
  • V. Thongboonkerd


Neutrophils have been thought to play a major role in inflammation and diabetic complications especially in poor glycemic control patients as demonstrated by their aberrant inflammatory markers. The aim of the present study was to compare neutrophil proteome profiles between diabetic patients with good glycemic control and those with poor glycemic control to see whether there might be any differences that could be related to the cause of complications which are found more commonly in the latter. Using 2-dimensional gel electrophoresis (2-DE) followed by quadrupole time of flight mass spectrometry (Q-TOF MS) and/or tandem mass spectrometry (MS/MS), we identified 35 differentially expressed proteins, some of which were protein components of neutrophil extracellular traps (NETs), in the poor glycemic control group compared to the good glycemic control group. The observed alterations of protein components of NETs included downregulation of myeloperoxidase, azurocidin (CAP37), and S100A9; and upregulation of the glycolytic enzymes transketolase and alpha-enolase. Manganese superoxide dismutase (MnSOD), functioning in cellular response and defense, was also found downregulated in the poor control group. Most of the glycolysis-related proteins were downregulated in the good control group but upregulated in the poor control group, including phosphoglycerate kinase 1 (PGK1) and L-lactate dehydrogenase B chain (LDHB). The findings of this study demonstrate the dysregulation of protein components of NETs in neutrophils in patients with poorly controlled diabetes. More specifically, these findings suggest association between NETs and inflammation in diabetes and provide further insights into the role of neutrophils in the complications of poorly controlled diabetes.


Diabetes Glycolytic enzymes Granular proteins Hyperglycemia Inflammation NETs Neutrophils 



Two-dimensional gel electrophoresis




Advanced glycation end product


Cationic antimicrobial protein of molecular weight 37 kDa


3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propanesulfonate


Diabetes mellitus


Deoxyribonucleic acid




Ethylenediaminetetraacetic acid


Glyceraldehyde-3-phosphate dehydrogenase


Hemoglobin A1c


High-density lipoprotein


Isoelectric focusing




l-Lactate dehydrogenase


l-Lactate dehydrogenase B chain


Low-density lipoprotein




Manganese superoxide dismutase




Mass spectrometry


Tandem mass spectrometry


Nicotinamide adenine dinucleotide phosphate


Neutrophil extracellular traps


Peripheral blood mononuclear cell


Phosphate-buffered saline


Phosphoglycerate kinase 1


Protein kinase C


Phorbol 12-myristate 13-acetate


Polymorphonuclear leukocyte


Quadrupole time of flight mass spectrometry


Reactive oxygen species


Sodiumdeodecyl sulfate polyacrylamide gel electrophoresis


Type 2 diabetes mellitus


Trifluoroacetic acid


Tumor necrosis factor



The authors gratefully acknowledge Kittisak Sintiprungrat for technical assistance. This study was supported by a Mahidol University Research Grant (Grant No. IRN60W0004) and a grant from the Thailand Research Fund (Grant No. IRG5980006). Umsa-ard W. was partially supported by a Siriraj Graduate Thesis Scholarship.

Author contributions

JS, WU, and VT conceived the study; JS and VT designed the experiments; WU performed the experiments; WU and VT analyzed the data; and, JS wrote the manuscript. All authors approved the final manuscript.

Compliance with ethical standards

Conflict of interest

All authors declare no personal or professional conflicts of interest, and no financial support from the companies that produce and/or distribute the drugs, devices, or materials described in this report.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research ethics committee (Siriraj Institutional Review Board COA No. Si 012/2008) and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Written informed consent was obtained from all participants.

Supplementary material

11010_2019_3583_MOESM1_ESM.docx (396 kb)
Supplementary material 1 (DOCX 395 kb)


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

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

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