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The lncRNA ROR/miR-124-3p/TRAF6 axis regulated the ischaemia reperfusion injury-induced inflammatory response in human cardiac myocytes

  • Ying-Ping Liang
  • Qin Liu
  • Guo-Hai Xu
  • Jing Zhang
  • Yong Chen
  • Fu-Zhou Hua
  • Chang-Qing Deng
  • Yan-Hui HuEmail author
Article

Abstract

Myocardial ischaemia reperfusion injury (MIRI) is considered the primary cause of death in patients with cardiovascular diseases. Recently, long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) have been found to be involved in the pathogenesis of MIRI. However, whether lncRNA ROR and miR-124-3p play roles in MIRI and the underlying mechanism remain undetermined. HCMs were exposed to hypoxic conditions for 2 h followed by re-oxygenation (H/R) treatment. Expression of miR-124-3p and lncRNA ROR in HCMs was measured by qRT-PCR. TRAF6 expression was evaluated by qRT-PCR and western blotting. ELISA and qRT-PCR were conducted to assess the production of TNF-α, IL-6, and IL-1β. The interaction between miR-124-3p and TRAF6, as well as between miR-124-3p and lncRNA ROR, was verified by dual-luciferase reporter assay. Cell apoptosis was detected by flow cytometry analysis. Our data revealed that miR-124-3p was significantly downregulated, while TRAF6 and lncRNA ROR were upregulated in both MIRI rat model and H/R treated HCMs. Overexpression of miR-124-3p reversed the H/R-induced cell apoptosis and upregulation of TNF-α, IL-6, and IL-1β. Mechanistically, miR-124-3p bound and negatively regulated TRAF6 expression in HCMs. Moreover, TRAF6 overexpression significantly blocked the effects of miR-124-3p mimics on cell apoptosis and inflammatory response of HCMs, which involved the NF-κB pathway. Further analysis showed that lncRNA ROR sponged and negatively regulated miR-124-3p in HCMs. Overexpression of IL-1β was demonstrated to promote H/R induced cell apoptosis in HCMs. In addition, overexpression of ROR further enhanced the H/R-induced inflammation and cell apoptosis through its action on miR-124-3p. The lncRNA ROR/miR-124-3p/TRAF6 axis regulated the H/R-induced cell apoptosis and inflammatory response of HCMs.

Keywords

LncRNA ROR miR-124-3p MIRI TRAF6 HCM 

Abbreviations

MIRI

Myocardial ischaemia reperfusion injury

LncRNA

Long non-coding RNAs

miRNA

Micro RNAs

IRI

Ischemia reperfusion injury

HCM

human cardiac myocytes

TRAF6

TNF receptor associated factor 6

CHD

Coronary heart disease

ELISA

Enzyme Linked Immunosorbent Assay

qRT-PCR

Quantitative real-time PCR

WT

Wild type

MUT

Mutant

LAD

Left anteriordescending coronaryartery

Notes

Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of AnesthesiologySecond Affiliated Hospital of Nanchang UniversityNanchangPeople’s Republic of China
  2. 2.Department of GastroenterologyThe Affiliated Hospital of Jiangxi University of Traditional Chinese MedicineNanchangPeople’s Republic of China

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