Elevated red blood cell‑derived extracellular vesicles under hyperglycemic conditions are associated with CD47 expression and production of intracellular reactive oxygen species

Sukati,Suriyan; Chunglok,Warangkana; Naulkaew,Tiwaporn; Bumm,Thomas; Jittivisuthikul,Surasak; Chan,Szn Yi; Wongsirojkul,Nichaporn; Wangmuang,Nantawan; Phiewkham,Duangjai; Prachongsai,Issara

doi:10.3892/br.2025.2007

Elevated red blood cell‑derived extracellular vesicles under hyperglycemic conditions are associated with CD47 expression and production of intracellular reactive oxygen species

Authors:
- Suriyan Sukati
- Warangkana Chunglok
- Tiwaporn Naulkaew
- Thomas Bumm
- Surasak Jittivisuthikul
- Szn Yi Chan
- Nichaporn Wongsirojkul
- Nantawan Wangmuang
- Duangjai Phiewkham
- Issara Prachongsai
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Affiliations: Department of Medical Technology, School of Allied Health Sciences, Walailak University, Nakhon Si Thammarat 80160, Thailand, Department of Internal Medicine II, Hematology and Oncology, University Clinic Würzburg, Würzburg D-97080, Germany, Department of Internal Medicine II, Hematology and Oncology, University Clinic Würzburg, Würzburg D-97080, Germany
Published online on: June 2, 2025 https://doi.org/10.3892/br.2025.2007
Article Number: 129
Copyright: © Sukati et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Diabetes mellitus (DM), particularly type 2 DM, is characterized by chronic hyperglycemia, which has notable effects on red blood cells (RBCs), including alterations in membrane integrity, morphology and function. The present study aimed to assess the effect of oxidative stress under hyperglycemic conditions on RBC‑derived extracellular vesicle (REV) production using an in vitro model. RBCs from healthy participants were cultured in several glucose concentrations (5, 45 and 100 mM) to mimic euglycemic, intermediate and severe hyperglycemic conditions. The levels of intracellular reactive oxygen species (ROS) and CD47‑expressing RBCs were measured using flow cytometry. REVs were purified using centrifugation and then their levels were measured using CountBright and flow cytometry. The results revealed a significantly increased level of intracellular ROS and REV production under hyperglycemic conditions. Additionally, hyperglycemia was demonstrated to reduce CD47 expression in RBCs, particularly at higher glucose concentrations. Elevated ROS production was also significantly associated with REV production in the early phase of hyperglycemic conditions, whereas the significant decrease in CD47 expression was observed later. Furthermore, pretreatment with N‑acetylcysteine effectively reduced ROS production and REV formation, highlighting the role of oxidative stress in hyperglycemia‑induced RBC alterations and REV production. The findings of the present study suggest that prolonged hyperglycemia facilitates intracellular oxidative stress and RBC membrane vesiculation, which markedly contributed to the understanding of the pathophysiology of diabetes‑related complications.

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