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Myricetin-induced suicidal erythrocyte death

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Abstract

Background

Myricetin, a type of flavonol commonly found in fruits and herbs, has demonstrated anticancer properties by triggering the process of apoptosis or programmed cell death in tumor cells. Despite the absence of mitochondria and nuclei, erythrocytes can undergo programmed cell death, also known as eryptosis.This process is characterized by cell shrinkage, externalization of phosphatidylserine (PS) on the cell membrane, and the formation of membrane blebs. The signaling of eryptosis involves Ca2+ influx, the formation of reactive oxygen species (ROS), and the accumulation of cell surface ceramide. The present study explored the effects of myricetin on eryptosis.

Methods and results

Human erythrocytes were exposed to various concentrations of myricetin (2–8 µM) for 24 h. Flow cytometry was used to assess the markers of eryptosis, including PS exposure, cellular volume, cytosolic Ca2+ concentration, and ceramide accumulation. In addition, the levels of intracellular ROS were measured using the 2’,7’-dichlorofluorescin diacetate (DCFDA) assay. The myricetin-treated (8 µM) erythrocytes significantly increased Annexin-positive cells, Fluo-3 fluorescence intensity, DCF fluorescence intensity, and the accumulation of ceramide. The impact of myricetin on the binding of annexin-V was significantly reduced, but not completely eliminated, by the nominal removal of extracellular Ca2+.

Conclusion

Myricetin triggers eryptosis, which is accompanied and, at least in part, caused by Ca2+ influx, oxidative stress and increase of ceramide abundance.

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Abbreviations

PS:

Phosphatidylserine

[Ca2+]i :

Cytosolic Ca2+ activity

ROS:

Reactive oxygen species

FSC:

Forward scatter

DCFDA:

2′,7′-dichlorodihydrofluorescein diacetate

DMSO:

Dimethyl sulfoxide

FITC:

Fluorescein isothiocyanate

EGTA:

Ethylenebis (oxyethylenenitrilo) tetraacetic acid

BSA:

Bovine serum albumin

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Acknowledgements

The authors acknowledge the meticulous preparation of the manuscript by Lejla Subasic.

Funding

This study was funded by China Postdoctoral Science Foundation (2021MD703799).

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Author notes

  1. Jibin Liu and Abdulla Al Mamun Bhuyan contributed equally to this work.

Authors and Affiliations

  1. College of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, 1166 Liutai Avenue, Wenjiang, Chengdu, 611137, People’s Republic of China

    Jibin Liu

  2. Department of Physiology, Eberhard-Karls-University of Tuebingen, Wilhlmstr. 56, 72076, Tuebingen, Germany

    Jibin Liu, Abdulla Al Mamun Bhuyan, Ke Ma, Xuexue Zhu, Kuo Zhou & Florian Lang

  3. Department of Veterinary and Animal Sciences, University of Rajshahi, Rajshahi, 6250, Bangladesh

    Abdulla Al Mamun Bhuyan

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  1. Jibin Liu
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Contributions

All authors contributed to the study conception and design. JL, AAMB and FL conceived and designed research. Material preparation, data collection and analysis were performed by JL, AAMB, KM, XZ and KZ. The first draft of the manuscript was written by FL and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Florian Lang.

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The study is approved by the ethics committee of Eberhard-Karls-University of Tuebingen (184/2003 V).

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Liu, J., Mamun Bhuyan, A.A., Ma, K. et al. Myricetin-induced suicidal erythrocyte death. Mol Biol Rep 50, 4253–4260 (2023). https://doi.org/10.1007/s11033-023-08350-3

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