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Heart and Vessels

, Volume 33, Issue 10, pp 1185–1194 | Cite as

Downregulation of miR-34a promotes endothelial cell growth and suppresses apoptosis in atherosclerosis by regulating Bcl-2

  • Gang Su
  • Guangli SunEmail author
  • Hai Liu
  • Liliang Shu
  • Zhenxing Liang
Original Article

Abstract

Several miRNAs have been demonstrated to be involved in endothelial dysfunction during atherosclerosis (AS). However, the detailed roles and underlying mechanisms of miR-34a in AS-associated endothelial cell apoptosis are far from being addressed. Apolipoprotein E-deficient (ApoE−/−) mice fed with high-fat diet (HFD) were used as in vivo model of AS. Oxidized low-density lipoprotein (ox-LDL)-treated human aortic endothelial cells (HAECs) were applied as in vitro model of AS. The effects of miR-34a on atherosclerotic lesions were evaluated by hematoxylin–eosin (HE) and Oil Red O staining. Pecam-1+ endothelial cells were isolated from the aortic arch with flow cytometry. qRT-PCR and western blot were employed to measure gene and protein expression. The effects of miR-34a on cell viability, cell cycle distribution, and apoptosis were assessed by Cell counting kit (CCK)-8 and flow cytometry analysis. The relationship between miR-34a and Bcl-2 was confirmed by online softwares, luciferase reporter assay, and RNA immunoprecipitation (RIP). miR-34a was upregulated in HFD-induced ApoE−/− mice and ox-LDL-treated HAECs. Anti-miR-34a decreased atherosclerotic lesions and inhibited Pecam-1+ endothelial cells apoptosis in HFD-induced ApoE−/− mice. Moreover, anti-miR-34a significantly promoted cell viability, alleviated cell cycle arrest, and restrained apoptosis in ox-LDL-treated HAECs. Furthermore, Bcl-2 was identified as a target of miR-34a, and miR-34a inhibited Bcl-2 expression via binding to its 3′UTR. Rescue experiments demonstrated that Bcl-2 overexpression dramatically reversed miR-34a-mediated inhibition of cell growth and promotion of apoptosis in ox-LDL-exposed HAECs. Depletion of miR-34a facilitated endothelial cell growth and blocked apoptosis in AS by upregulating Bcl-2, offering a promising avenue for AS therapy.

Keywords

Atherosclerosis ApoE−/− mice Endothelial cells miR-34a Bcl-2 High-fat diet Oxidized low-density lipoprotein 

Notes

Acknowledgements

This work was supported by Henan Provincial Department of Education research project (No. 162102310550).

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest to declare.

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Copyright information

© Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Cardiac SurgeryThe First Affiliated Hospital of Zhengzhou UniversityZhengzhouChina
  2. 2.Department of OphthalmologyThe First Affiliated Hospital of Zhengzhou UniversityZhengzhouChina

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