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

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A Correction to this article was published on 07 April 2021

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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.

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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

References

  • Anfossi S, Fu X, Nagvekar R, Calin GA (2018) MicroRNAs, regulatory messengers inside and outside Cancer cells. Adv Exp Med Biol 1056:87–108

    Article  CAS  Google Scholar 

  • Chen X, Sun Y, Cai R, Wang G, Shu X, Pang W (2018) Long noncoding RNA: multiple players in gene expression. BMB Rep 51(6):280–289

    Article  CAS  Google Scholar 

  • Chen PJ et al (2018) microRNA-874 inhibition targeting STAT3 protects the heart from ischemia-reperfusion injury by attenuating cardiomyocyte apoptosis in a mouse model. J Cell Physiol

  • Dey BK, Mueller AC, Dutta A (2014) Long non-coding RNAs as emerging regulators of differentiation, development, and disease. Transcription 5(4):e944014

    Article  Google Scholar 

  • Frank A, Bonney M, Bonney S, Weitzel L, Koeppen M, Eckle T (2012) Myocardial ischemia reperfusion injury: from basic science to clinical bedside. Semin Cardiothorac Vasc Anesth 16(3):123–132

    Article  Google Scholar 

  • Giroud M, Scheideler M (2017) Long Non-Coding RNAs in Metabolic Organs and Energy Homeostasis. Int J Mol Sci 18(12)

  • Gong Q, Su G (2017) Roles of miRNAs and long noncoding RNAs in the progression of diabetic retinopathy. Biosci Rep 37(6)

  • Gonzalez-Montero J et al (2018) Myocardial reperfusion injury and oxidative stress: therapeutic opportunities. World J Cardiol 10(9):74–86

    Article  Google Scholar 

  • Hausenloy DJ, Yellon DM (2013) Myocardial ischemia-reperfusion injury: a neglected therapeutic target. J Clin Invest 123(1):92–100

    Article  CAS  Google Scholar 

  • Huang Y (2018) The novel regulatory role of lncRNA-miRNA-mRNA axis in cardiovascular diseases. J Cell Mol Med 22(12):5768–5775

    Article  CAS  Google Scholar 

  • Huang S, Ge X, Yu J, Han Z, Yin Z, Li Y, Chen F, Wang H, Zhang J, Lei P (2018) Increased miR-124-3p in microglial exosomes following traumatic brain injury inhibits neuronal inflammation and contributes to neurite outgrowth via their transfer into neurons. FASEB J 32(1):512–528

    Article  CAS  Google Scholar 

  • Ishida T, Mizushima Si, Azuma S, Kobayashi N, Tojo T, Suzuki K, Aizawa S, Watanabe T, Mosialos G, Kieff E, Yamamoto T, Inoue J (1996) Identification of TRAF6, a novel tumor necrosis factor receptor-associated factor protein that mediates signaling from an amino-terminal domain of the CD40 cytoplasmic region. J Biol Chem 271(46):28745–28748

    Article  CAS  Google Scholar 

  • Jang YH, Kim JH, Ban C, Ahn K, Cheong JH, Kim HH, Kim JS, Park YH, Kim J, Chun KJ, Lee GH, Kim M, Kim C, Xu Z (2012) Stromal cell derived factor-1 (SDF-1) targeting reperfusion reduces myocardial infarction in isolated rat hearts. Cardiovasc Ther 30(5):264–272

    Article  CAS  Google Scholar 

  • Jiang W et al (2014) miR-146a ameliorates liver ischemia/reperfusion injury by suppressing IRAK1 and TRAF6. PLoS One 9(7):e101530

    Article  Google Scholar 

  • Kong L, Sun M, Jiang Z, Li L, Lu B (2018) MicroRNA-194 inhibits lipopolysaccharide-induced inflammatory response in nucleus Pulposus cells of the intervertebral disc by targeting TNF receptor-associated factor 6 (TRAF6). Med Sci Monit 24:3056–3067

    Article  CAS  Google Scholar 

  • Lejay A, Fang F, John R, van J, Barr M, Thaveau F, Chakfe N, Geny B, Scholey JW (2016) Ischemia reperfusion injury, ischemic conditioning and diabetes mellitus. J Mol Cell Cardiol 91:11–22

    Article  CAS  Google Scholar 

  • Li C, Zhao Z, Zhou Z, Liu R (2016) Linc-ROR confers gemcitabine resistance to pancreatic cancer cells via inducing autophagy and modulating the miR-124/PTBP1/PKM2 axis. Cancer Chemother Pharmacol 78(6):1199–1207

    Article  CAS  Google Scholar 

  • Liu F, Hu H, Zhao J, Zhang Z, Ai X, Tang L, Xie L (2018) miR-124-3p acts as a potential marker and suppresses tumor growth in gastric cancer. Biomed Rep 9(2):147–155

    CAS  PubMed  PubMed Central  Google Scholar 

  • Loewer S, Cabili MN, Guttman M, Loh YH, Thomas K, Park IH, Garber M, Curran M, onder T, Agarwal S, Manos PD, Datta S, Lander ES, Schlaeger TM, Daley GQ, Rinn JL (2010) Large intergenic non-coding RNA-RoR modulates reprogramming of human induced pluripotent stem cells. Nat Genet 42(12):1113–1117

    Article  CAS  Google Scholar 

  • Long HD et al (2018) Reduced hsa-miR-124-3p levels are associated with the poor survival of patients with hepatocellular carcinoma. Mol Biol Rep

  • Lu R et al (2018) Prognostic value of lncRNA ROR expression in various cancers: a meta-analysis. Biosci Rep 38(5)

  • Ong SB, Katwadi K, Kwek XY, Ismail NI, Chinda K, Ong SG, Hausenloy DJ (2018) Non-coding RNAs as therapeutic targets for preventing myocardial ischemia-reperfusion injury. Expert Opin Ther Targets 22(3):247–261

    Article  CAS  Google Scholar 

  • Qian Z, Zhou S, Zhou Z, Yang X, Que S, Lan J, Qiu Y, Lin Y (2017) miR146b5p suppresses glioblastoma cell resistance to temozolomide through targeting TRAF6. Oncol Rep 38(5):2941–2950

    Article  CAS  Google Scholar 

  • Quinn JJ, Chang HY (2016) Unique features of long non-coding RNA biogenesis and function. Nat Rev Genet 17(1):47–62

    Article  CAS  Google Scholar 

  • Rauf A et al (2019) The role of Caspase 1 in ischemia/reperfusion injury of the myocardium. J Cardiovasc Pharmacol

  • Sun W et al (2019) Gastrodin ameliorates microvascular reperfusion injury-induced pyroptosis by regulating the NLRP3/caspase-1 pathway. J Physiol Biochem

  • Tan H, Qi J, Fan BY, Zhang J, Su FF, Wang HT (2018) MicroRNA-24-3p attenuates myocardial ischemia/reperfusion injury by suppressing RIPK1 expression in mice. Cell Physiol Biochem 51(1):46–62

    Article  CAS  Google Scholar 

  • Tang LX, Chen GH, Li H, He P, Zhang Y, Xu XW (2018) Long non-coding RNA OGFRP1 regulates LYPD3 expression by sponging miR-124-3p and promotes non-small cell lung cancer progression. Biochem Biophys Res Commun 505(2):578–585

    Article  CAS  Google Scholar 

  • Uchida S, Dimmeler S (2015) Long noncoding RNAs in cardiovascular diseases. Circ Res 116(4):737–750

    Article  CAS  Google Scholar 

  • Walsh MC, Lee J, Choi Y (2015) Tumor necrosis factor receptor- associated factor 6 (TRAF6) regulation of development, function, and homeostasis of the immune system. Immunol Rev 266(1):72–92

    Article  CAS  Google Scholar 

  • Wang X, Ha T, Liu L, Zou J, Zhang X, Kalbfleisch J, Gao X, Williams D, Li C (2013) Increased expression of microRNA-146a decreases myocardial ischaemia/reperfusion injury. Cardiovasc Res 97(3):432–442

    Article  CAS  Google Scholar 

  • Wang X, Ha T, Zou J, Ren D, Liu L, Zhang X, Kalbfleisch J, Gao X, Williams D, Li C (2014) MicroRNA-125b protects against myocardial ischaemia/reperfusion injury via targeting p53-mediated apoptotic signalling and TRAF6. Cardiovasc Res 102(3):385–395

    Article  CAS  Google Scholar 

  • Wang K, Liu F, Liu CY, An T, Zhang J, Zhou LY, Wang M, Dong YH, Li N, Gao JN, Zhao YF, Li PF (2016) The long noncoding RNA NRF regulates programmed necrosis and myocardial injury during ischemia and reperfusion by targeting miR-873. Cell Death Differ 23(8):1394–1405

    Article  CAS  Google Scholar 

  • Wang HB, Yang J, Ding JW, Chen LH, Li S, Liu XW, Yang CJ, Fan ZX, Yang J (2016) RNAi-mediated Down-regulation of CD47 protects against ischemia/reperfusion-induced myocardial damage via activation of eNOS in a rat model. Cell Physiol Biochem 40(5):1163–1174

    Article  CAS  Google Scholar 

  • Wei C, Lei L, Hui H, Tao Z (2019) MicroRNA-124 regulates TRAF6 expression and functions as an independent prognostic factor in colorectal cancer. Oncol Lett 18(1):856–863

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang W, Li Y, Wang P (2018) Long non-coding RNA-ROR aggravates myocardial ischemia/reperfusion injury. Braz J Med Biol Res 51(6):e6555

    Article  Google Scholar 

  • Zhang W, Zhao H, Wu Q, Xu W, Xia M (2018) Knockdown of BACE1-AS by siRNA improves memory and learning behaviors in Alzheimer's disease animal model. Exp Ther Med 16(3):2080–2086

    PubMed  PubMed Central  Google Scholar 

  • Zhao C et al (2019) Up-regulation of ANXA1 suppresses polymorphonuclear neutrophil infiltration and myeloperoxidase activity by activating STAT3 signaling pathway in rat models of myocardial ischemia reperfusion injury. Cell Signal

  • Zheng C et al (2018) Long noncoding RNA AK12348 is involved in the regulation of myocardial Ischaemia-reperfusion injury by targeting PARP and Caspase-3. Heart Lung Circ 27(5):e51–e58

    Article  Google Scholar 

  • Zo RB, Long Z (2018) MiR-124-3p suppresses bladder cancer by targeting DNA methyltransferase 3B. J Cell Physiol 234(1):464–474

    Article  Google Scholar 

Download references

Acknowledgements

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

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Authors and Affiliations

  1. Department of Anesthesiology, Second Affiliated Hospital of Nanchang University, No.1, Minde Road, Nanchang, 330006, Jiangxi Province, People’s Republic of China

    Ying-Ping Liang, Qin Liu, Guo-Hai Xu, Jing Zhang, Yong Chen, Fu-Zhou Hua & Yan-Hui Hu

  2. Department of Gastroenterology, The Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang, 330006, Jiangxi Province, People’s Republic of China

    Chang-Qing Deng

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  1. Ying-Ping Liang
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The original version of this article unfortunately a mistake. The primer sequences of TRAF6 in “Quantitative real-time PCR (qRT-PCR) assay” of the Method section should be changed from TRAF6: forward 5’-CAG TGG TCG TAT CGT GCT TA-3′, reverse 5’-CCT TAT GGT TTC TTG GAG TC-3′ to TRAF6: forward 5’-TCG AAC CCT TGA GGA CAA AG -3’, reverse 5’- CGG GTT TGC CAG TGT AGA AT-3’. The original article has been corrected.

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Liang, YP., Liu, Q., Xu, GH. et al. The lncRNA ROR/miR-124-3p/TRAF6 axis regulated the ischaemia reperfusion injury-induced inflammatory response in human cardiac myocytes. J Bioenerg Biomembr 51, 381–392 (2019). https://doi.org/10.1007/s10863-019-09812-9

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