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Extracellular Vesicles Derived from Intermittent Hypoxia–Treated Red Blood Cells Impair Endothelial Function Through Regulating eNOS Phosphorylation and ET-1 Expression

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Abstract

Purpose

Intermittent hypoxia (IH), a main characteristic of obstructive sleep apnea (OSA) syndrome, has been known as a dominant cause of OSA-related endothelial dysfunction and hypertension. However, the underlying mechanism still remains unclear. Extracellular vesicles (EVs), small vesicles secreted by various cells, can be absorbed by endothelial cells and then influence vascular function. The aim of this research is to clarify whether and how EVs shedding from red blood cells (RBCs) are involved in IH-induced endothelial dysfunction.

Methods

EVs were extracted by ultracentrifugation. After the identification of property and purity, EVs from IH-exposed RBCs (IH REVs) and normoxia-exposed RBCs (NOR REVs) or from OSA and non-OSA patient RBCs were utilized to treat C57BL/6 mouse aortas or human umbilical vein endothelial cells (HUVECs) for mechanistic exploration.

Results

Functional results demonstrated that REVs from OSA patients dramatically impaired endothelium-dependent relaxations (EDRs). Similarly, in vivo and ex vivo studies showed that IH REVs caused significant endothelial dysfunction compared to control group. Further results presented that IH REVs blocked endothelial nitric oxide synthase (eNOS) phosphorylation through inhibiting PI3K/Akt pathway and enhanced endothelin-1 (ET-1) expression through activating Erk1/2 pathway in endothelial cells. Meanwhile, endothelial dysfunction caused by IH REVs was reversed by Akt activator SC79 as well as Erk kinase inhibitor PD98059, suggesting that PI3K/Akt/eNOS and Erk1/2/ET-1 pathways were implicated in IH REV-induced impaired EDRs.

Conclusions

This study reveals a novel role of REVs in endothelial dysfunction under IH and dissects the relevant mechanism involved in this process, which will help to establish a comprehensive understanding of OSA or IH-related endothelial dysfunction from a new scope.

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Acknowledgments

We are grateful to Ms. Yuqing Yang (Bei**g ECHO Biotech. Co., Ltd. China) for transmission electron microscopy assistance.

Funding

This study was supported by Bei**g Natural Science Foundation under Grant No. 7192031, by National Natural Science Foundation of China under Grant No. 81870335, and by Bei**g Key Laboratory of Upper Airway Dysfunction and Related Cardiovascular Diseases under Grant No. BZ0377.

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Authors and Affiliations

  1. Key Laboratory of Upper Airway Dysfunction-related Cardiovascular Diseases, Bei**g Institute of Heart, Lung and Blood Vessel Disease, Bei**g Anzhen Hospital, Capital Medical University, No. 2 Anzhen Road, Bei**g, 100029, China

    Lu Peng, Yu Li, **nwei Li, Yunhui Du, Linyi Li, Chaowei Hu, **g Zhang, Yanwen Qin & Huina Zhang

  2. Department of Otolaryngology-Head and Neck Surgery, Bei**g Anzhen Hospital, Capital Medical University, No. 2 Anzhen Road, Bei**g, 100029, China

    Yongxiang Wei

Authors
  1. Lu Peng
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  2. Yu Li
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  3. **nwei Li
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  4. Yunhui Du
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  5. Linyi Li
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  6. Chaowei Hu
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  7. **g Zhang
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  8. Yanwen Qin
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  10. Huina Zhang
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Contributions

Lu Peng carried out main experiments, designed some experiments, and wrote the manuscript. Yu Li and **nwei Li helped to do the animal experiments. Yunhui Du, Linyi Li, and Chaowei Hu helped to analyze the data. **g Zhang and Yanwen Qin helped with sample collection. Huina Zhang and Yongxiang Wei conceived the experiments, edited the manuscript, and financed the study.

Corresponding authors

Correspondence to Yongxiang Wei or Huina Zhang.

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All procedures involving human participants were authorized by the Medical Ethics Committee of Bei**g Anzhen Hospital and adhered to the Declaration of Helsinki and its later amendments or comparable ethical standards. All procedures involving animals were approved by Capital Medical University Animal Experimentation Ethics Committee and carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Peng, L., Li, Y., Li, X. et al. Extracellular Vesicles Derived from Intermittent Hypoxia–Treated Red Blood Cells Impair Endothelial Function Through Regulating eNOS Phosphorylation and ET-1 Expression. Cardiovasc Drugs Ther 35, 901–913 (2021). https://doi.org/10.1007/s10557-020-07117-3

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