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Blocking C/EBP β protects vascular endothelial cells from injury induced by intermittent hypoxia

  • Basic Science • Original Article
  • Published:
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Abstract

Background

Intermittent hypoxia (IH) can damage endothelial cells and lead to apoptosis in obstructive sleep apnea-hypopnea syndrome (OSAHS). Hypoxia induces apoptosis in endothelial cells via upregulation of endothelin-1 (ET-1) and hypoxia inducible factor-1 alpha (HIF-1α) plays a key role in the hypoxic stress response.

Purpose

We investigated an approach to diminish the negative effect of HIF-1α while maintaining its protective effect.

Methods

Human umbilical vein endothelial cells (HUVECs) were subjected to sustained hypoxia (SH) or IH for 24 h, and the responses of HIF-1α, CCAAT/enhancer binding protein beta (C/EBP β), and endothelin-1 (ET-1) were assessed by western blotting. A luciferase reporter system was employed to verify the potential binding site (transcription factor binding site, TFBS) for C/EBP β in the ET-1 promoter. The specificity of regulation of ET-1 by HIF-1α via C/EBP β was evaluated by a lentiviral system. The effects of silencing of C/EBP β on IH-induced apoptosis, vascular endothelial growth factor (VEGF) protein levels, proliferation, and in vitro tube formation were studied.

Results

We found that IH significantly increased HIF-1α, C/EBP β, and ET-1 in HUVECs. Knockdown of HIF-1α or C/EBP β inhibited the upregulation of ET-1 induced by IH. Blocking C/EBP β impaired IH-induced apoptosis but did not affect VEGF expression, proliferation, or in vitro tube formation. C/EBP β was shown to mediate increased ET-1 transcription by HIF-1α through the TFBS, 5′-GTTGCCTGTTG-3′, in ET-1 promoter.

Conclusion

Silencing of C/EBP β can suppress apoptosis but does not affect the protective role of HIF-1α in the hypoxic stress response.

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Abbreviations

SH:

Sustained hypoxia

IH:

Intermittent hypoxia

OSAHS:

Obstructive sleep apnea-hypopnea syndrome

HIF-1α:

Hypoxia-inducible factor 1-alpha

C/EBP β:

CCAAT/enhancer binding protein beta

HUVECs:

Human umbilical vein endothelial cells

ET-1:

Endothelin-1

VEGF:

Vascular endothelial growth factor

TFBS:

Transcription factor binding site

FBS:

Fetal bovine serum

MOI:

Multiplicity of infection

TBST:

Tris-buffered saline with Tween 20

PVDF:

Polyvinylidene fluoride

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Funding

The National Natural Science Foundation of China provided financial support in the form of national natural science funding (No. 81372529), the Natural Science Fund Project of Shanghai (13ZR1414400); the Innovation Fund Project of Education Commission in Shanghai (14ZZ079); and the Special issue of the Military Medical of Second Military Medical University (2012JS21). The sponsor had no role in the design or conduct of this research.

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Author notes

    Authors and Affiliations

    1. Department of Respiratory Medicine, Shanghai East Hospital, Tongji University School of Medicine, Pudong, Shanghai, 200120, China

      Yu Feng & Liming Zhao

    2. Department of Radiology, Secondary Affiliated Hospital of the Second Military Medical University, Shanghai, 200001, China

      Qingchu Li

    3. Department of Respiratory Medicine, Third Affiliated Hospital of Second Military Medical University, Shanghai, 200438, China

      Yinxiang Wu, Nana Zhao & Lu Li

    4. Department of Teaching Support, Second Military Medical University, Shanghai, 200433, China

      Li Li

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    1. Yu Feng
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    Correspondence to Li Li or Liming Zhao.

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    Yu Feng and Qingchu Li contributed equally to this work.

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    Feng, Y., Li, Q., Wu, Y. et al. Blocking C/EBP β protects vascular endothelial cells from injury induced by intermittent hypoxia. Sleep Breath 23, 953–962 (2019). https://doi.org/10.1007/s11325-018-1759-7

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