The MicroRNA-210/Casp8ap2 Pathway Alleviates Hypoxia-Induced Injury in Myocardial Cells by Regulating Apoptosis and Autophagy
DOI:
https://doi.org/10.1532/hsf.3173Keywords:
miR-210; Caspase8ap2; myocardial injury; hypoxia; apoptosis; autophagyAbstract
Aim: This study was conducted to investigate the role of the miR-210/Caspase8ap2 pathway in apoptosis and autophagy in hypoxic myocardial cells.
Methods: The miR-control, miR-210 mimic, and miR-210 inhibitor were transfected into rat myocardial H9C2 cells. The transfection efficiency of exogenous miR-210 was determined by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). H9C2 cells were then treated with CoCl2 for 24, 48, and 72 h to generate a myocardial injury model. The apoptosis of H9C2 cells was assessed by flow cytometry. Additionally, a western blot assay was used to determine the expression of the autophagy-associated proteins light chain 3 (LC3), p62 and Beclin-1, and apoptosis-associated proteins Caspase8ap2, cleaved caspase 8, and cleaved caspase 3.
Results: We determined that a 48 h hypoxia treatment duration in H9C2 cardiomyocytes induced myocardial injury. Additionally, the overexpression of miR-210 significantly inhibited cell apoptosis. MiR-210 suppressed autophagy by upregulating p62 and downregulating LC3II/I in hypoxic H9C2 cells. Caspase8ap2 was a putative target of miR-210, miR-210 mediated apoptosis, and autophagy of H9C2 cells via suppressing Caspase8ap2. Furthermore, the expression of caspase 8, caspase 3, and Beclin-1 were decreased in response to miR-210.
Conclusion: miR-210 exhibits anti-apoptosis and anti-autophagy effects, which alleviate myocardial injury in response to hypoxia.
References
Feng J, Huang S, He M, et al. 2014. Analysis on the association of single nucleotide polymorphism in the promoter region of pre-miR-320b-2 with coronary heart disease risk and factors influencing circulating microRNA-320b level. Zhonghua Yu Fang Yi Xue Za Zhi. 48:893-9.
Gou D, Ramchandran R, Peng X, et al. 2012. miR-210 has an antiapoptotic effect in pulmonary artery smooth muscle cells during hypoxia. Am J Physiol Lung Cell Mol Physiol 303: L682-91.
Greenwood JP, Maredia N, Younger JF, et al. 2012. Cardiovascular magnetic resonance and single-photon emission computed tomography for diagnosis of coronary heart disease (CE-MARC): a prospective trial. Lancet 379:453-60.
Hamacher-Brady A, Brady NR,Gottlieb RA. 2006. The interplay between pro-death and pro-survival signaling pathways in myocardial ischemia/reperfusion injury: apoptosis meets autophagy. Cardiovasc Drugs and Therapy 20:445-62.
He B, Zhao Y, Xu L, et al. 2016. The nuclear melatonin receptor RORalpha is a novel endogenous defender against myocardial ischemia/reperfusion injury. J Pineal Res 60:313-26.
Heran BS, Chen JM, Ebrahim S, et al. 2011. Exercise-based cardiac rehabilitation for coronary heart disease. Cochrane Database Syst Rev 7:CD001800.
Hu S, Huang M, Li Z, et al. 2010. MicroRNA-210 as a novel therapy for treatment of ischemic heart disease. Circulation 122(11 Suppl):S124-31.
Huang X, Ding L, Bennewith KL, et al. 2009. Hypoxia-inducible mir-210 regulates normoxic gene expression involved in tumor initiation. Mol Cell 35:856-67.
Huang X, Le QT, Giaccia AJ. 2010. MiR-210--micromanager of the hypoxia pathway. Trends Mol Med 16:230-7.
Huang X, Zuo J. 2014. Emerging roles of miR-210 and other non-coding RNAs in the hypoxic response. Acta Biochim Biophys Sin (Shanghai). 46:220-32.
Joubert PE, Werneke SW, de la Calle C, et al. 2012. Chikungunya virus-induced autophagy delays caspase-dependent cell death. J Exp Med 209:1029-47.
Kim HW, Haider HK, Jiang S, Ashraf M. 2009. Ischemic preconditioning augments survival of stem cells via miR-210 expression by targeting caspase-8-associated protein 2. J Biol Chem 284:33161-8.
Kim HW, Mallick F, Durrani S, Ashraf M, Jiang S, Haider KH. 2012. Concomitant activation of miR-107/PDCD10 and hypoxamir-210/Casp8ap2 and their role in cytoprotection during ischemic preconditioning of stem cells. Antioxid Redox Signal 17:1053-65.
Kuo HF, Liu PL, Chong IW, et al. 2016. Pigment epithelium-derived factor mediates autophagy and apoptosis in myocardial hypoxia/reoxygenation injury. PLoS One 11:e0156059.
Li R, Geng HH, Xiao J, et al. 2016. miR-7a/b attenuates post-myocardial infarction remodeling and protects H9c2 cardiomyoblast against hypoxia-induced apoptosis involving Sp1 and PARP-1. Sci Rep 6:29082.
Liang W, Guo J, Li J, Bai C, Dong Y. 2016. Downregulation of miR-122 attenuates hypoxia/reoxygenation (H/R)-induced myocardial cell apoptosis by upregulating GATA-4. Biochem Biophys Res Commun. 478:1416-22.
Liu H, Huan L, Yin J, et al. 2017. Role of microRNA-130a in myocardial hypoxia/reoxygenation injury. Exp Ther Med 13:759-65.
Ma S, Wang Y, Chen Y,Cao F. 2015. The role of the autophagy in myocardial ischemia/reperfusion injury. Biochimica et biophysica acta 1852:271-6.
Miller CL, Haas U, Diaz R, et al. 2014. Coronary heart disease-associated variation in TCF21 disrupts a miR-224 binding site and miRNA-mediated regulation. PLoS Genet 10:e1004263.
Muhammad N, Bhattacharya S, Steele R, Ray RB. 2016. Anti-miR-203 suppresses ER-positive breast cancer growth and stemness by targeting SOCS3. Oncotarget 7:58595-605.
Mutharasan RK, Nagpal V, Ichikawa Y, Ardehali H. 2011. microRNA-210 is upregulated in hypoxic cardiomyocytes through Akt- and p53-dependent pathways and exerts cytoprotective effects. Am J Physiol Heart Circ Physiol 301:H1519-30.
Nabialek E, Wanha W, Kula D, et al. 2013. Circulating microRNAs (miR-423-5p, miR-208a and miR-1) in acute myocardial infarction and stable coronary heart disease. Minerva Cardioangiol 61:627-37.
Nie Y, Han BM, Liu XB, et al. 2011. Identification of MicroRNAs involved in hypoxia- and serum deprivation-induced apoptosis in mesenchymal stem cells. Int J Biol Sci 7:762-8.
Pu J, Yuan A, Shan P, et al. 2013. Cardiomyocyte-expressed farnesoid-X-receptor is a novel apoptosis mediator and contributes to myocardial ischaemia/reperfusion injury. Eur Heart J 34:1834-45.
Rastam S, Al Ali R, Maziak W, et al. 2012. Explaining the increase in coronary heart disease mortality in Syria between 1996 and 2006. BMC Public Health 12:754.
Shimono Y, Mukohyama J, Nakamura S, Minami H. 2015. MicroRNA regulation of human breast cancer stem cells. J Clin Med 5:pii:E2.
Yang W, Sun T, Cao J, Liu F, Tian Y, Zhu W. 2012. Downregulation of miR-210 expression inhibits proliferation, induces apoptosis and enhances radiosensitivity in hypoxic human hepatoma cells in vitro. Exp Cell Res 318:944-54.
