MicroRNA-128 inhibition attenuates myocardial ischemia/reperfusion injury-induced cardiomyocyte apoptosis by the targeted activation of peroxisome proliferator-activated receptor gamma

Zeng,Xiao  Cong; Li,Lang; Wen,Hong; Bi,Qi

doi:10.3892/mmr.2016.5208

MicroRNA-128 inhibition attenuates myocardial ischemia/reperfusion injury-induced cardiomyocyte apoptosis by the targeted activation of peroxisome proliferator-activated receptor gamma

Authors:
- Xiao Cong Zeng
- Lang Li
- Hong Wen
- Qi Bi
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Affiliations: Department of Cardiology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
Published online on: May 4, 2016 https://doi.org/10.3892/mmr.2016.5208
Pages: 129-136
Copyright: © Zeng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the effects of microRNA (miR)-128 inhibition on the targeted activation of peroxisome proliferator-activated receptor gamma (PPARG) and on cardiomyocyte apoptosis induced by myocardial ischemia/reperfusion (I/R) injury. In vitro, the expression of PPARG was detected by reverse transcription-quantitative polymerase chain reaction and western blotting in neonatal rat ventricular myocytes (NRVMs) and HEK293 cells transfected with the mimics or inhibitors of miR‑128 or control RNA. Luciferase reporter assays were used to identify whether PPARG is a direct target of miR‑128. In vivo, miR‑128 was knocked down via ear vein injection of antagomir‑128 in a rabbit myocardial I/R injury model. Western blotting investigated the activation of Akt [phosphorylated (p)‑Akt] and the expression of total‑Akt, PPARG and myeloid leukemia cell differentiation protein‑1 (Mcl‑1) in the myocardium. Cardiomyocyte apoptosis was examined with transmission electron microscropy and terminal deoxynucleotidyl transferase dUTP nick end labeling staining. PPARG mRNA and protein were downregulated in NRVMs transfected with miR‑128 mimics, but upregulated by antagomir‑128 compared with control. This indicates that PPARG is a direct miR‑128 target. Activation of Akt (p‑Akt), Mcl‑1 and PPARG expression in the myocardium were increased by miR‑128 inhibition. Furthermore, miR‑128 antagomirs significantly reduced apoptosis in hearts subjected to I/R injury, which was blocked by the PPARG inhibitor GW9662. In conclusion, miR‑128 inhibition attenuated I/R injury‑induced cardiomyocyte apoptosis by the targeted activation of PPARG signaling.

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1	Canadian Cardiovascular Society American Academy of Family Physicians American College of Cardiology; American Heart Association; Antman EM, Hand M, Armstrong PW, Bates ER, Green LA, Halasyamani LK, et al: 2007 focused update of the ACC/AHA 2004 guidelines for the management of patients with ST-elevation myocardial infarction: A report of the American college of cardiology/American heart association task force on practice guidelines. J Am Coll Cardiol. 51:210–247. 2008. View Article : Google Scholar : PubMed/NCBI
2	Stenestrand U, Lindbäck J and Wallentin L: RIKS-HIA Registry: Long-term outcome of primary percutaneous coronary intervention vs. prehospital and in-hospital thrombolysis for patients with ST-elevation myocardial infarction. JAMA. 296:1749–1756. 2006. View Article : Google Scholar : PubMed/NCBI
3	Buja LM: Myocardial ischemia and reperfusion injury. Cardiovasc Pathol. 14:170–175. 2005. View Article : Google Scholar : PubMed/NCBI
4	Takemura G, Nakagawa M, Kanamori H, Minatoguchi S and Fujiwara H: Benefits of reperfusion beyond infarct size limitation. Cardiovasc Res. 83:269–276. 2009. View Article : Google Scholar : PubMed/NCBI
5	Baines CP: How and when do myocytes die during ischemia and reperfusion: The late phase. J Cardiovasc Pharmacol Ther. 16:239–243. 2011. View Article : Google Scholar : PubMed/NCBI
6	Eefting F, Rensing B, Wigman J, Pannekoek WJ, Liu WM, Cramer MJ, Lips DJ and Doevendans PA: Role of apoptosis in reperfusion injury. Cardiovasc Res. 61:414–426. 2004. View Article : Google Scholar : PubMed/NCBI
7	Jozkowicz A, Dulak J, Piatkowska E, Placha W and Dembinska-Kiec A: Ligands of peroxisome proliferator-activated receptor-gamma increase the generation of vascular endothelial growth factor in vascular smooth muscle cells and in macrophages. Acta Biochim Pol. 47:1147–1157. 2000.
8	Mersmann J, Tran N, Zacharowski PA, Grotemeyer D and Zacharowski K: Rosiglitazone is cardioprotective in a murine model of myocardial I/R. Shock. 30:64–68. 2008.PubMed/NCBI
9	Goyal S, Arora S, Bhatt TK, Das P, Sharma A, Kumari S and Arya DS: Modulation of PPAR-gamma by telmisartan protects the heart against myocardial infarction in experimental diabetes. Chem Biol Interact. 185:271–280. 2010. View Article : Google Scholar : PubMed/NCBI
10	Yasuda S, Kobayashi H, Iwasa M, Kawamura I, Sumi S, Narentuoya B, Yamaki T, Ushikoshi H, Nishigaki K, Nagashima K, et al: Antidiabetic drug pioglitazone protects the heart via activation of PPAR-gamma receptors, PI3-kinase, Akt and eNOS pathway in a rabbit model of myocardial infarction. Am J Physiol Heart Circ Physiol. 296:H1558–H1565. 2009. View Article : Google Scholar : PubMed/NCBI
11	Kilter H, Werner M, Roggia C, Reil JC, Schäfers HJ, Kintscher U and Böhm M: The PPAR-gamma agonist rosiglitazone facilitates Akt rephosphorylation and inhibits apoptosis in cardiomyocytes during hypoxia/reoxygenation. Diabetes Obes Metab. 11:1060–1067. 2009. View Article : Google Scholar : PubMed/NCBI
12	Ye Y, Hu Z, Lin Y, Zhang C and Perez-Polo JR: Downregulation of microRNA-29 by antisense inhibitors and a PPAR-gamma agonist protects against myocardial ischaemia-reperfusion injury. Cardiovasc Res. 87:535–544. 2010. View Article : Google Scholar : PubMed/NCBI
13	Ou HC, Lee WJ, Lee SD, Huang CY, Chiu TH, Tsai KL, Hsu WC and Sheu WH: Ellagic acid protects endothelial cells from oxidized low-density lipoprotein-induced apoptosis by modulating the PI3K/Akt/eNOS pathway. Toxicol Appl Pharmacol. 248:134–143. 2010. View Article : Google Scholar : PubMed/NCBI
14	Mott JL, Kobayashi S, Bronk SF and Gores GJ: mir-29 regulates Mcl-1 protein expression and apoptosis. Oncogene. 26:6133–6140. 2007. View Article : Google Scholar : PubMed/NCBI
15	Hullinger TG, Montgomery RL, Seto AG, Dickinson BA, Semus HM, Lynch JM, Dalby CM, Robinson K, Stack C, Latimer PA, et al: Inhibition of miR-15 protects against cardiac ischemic injury. Circ Res. 110:71–81. 2012. View Article : Google Scholar :
16	Lin Q, Gao Z, Alarcon RM, Ye J and Yun Z: A role of miR-27 in the regulation of adipogenesis. FEBS J. 276:2348–2358. 2009. View Article : Google Scholar : PubMed/NCBI
17	Wang J, Song Y, Zhang Y, Xiao H, Sun Q, Hou N, Guo S, Wang Y, Fan K, Zhan D, et al: Cardiomyocyte overexpression of miR-27b induces cardiac hypertrophy and dysfunction in mice. Cell Res. 22:516–527. 2012. View Article : Google Scholar :
18	Lee EK, Lee MJ, Abdelmohsen K, Kim W, Kim MM, Srikantan S, Martindale JL, Hutchison ER, Kim HH, Marasa BS, et al: miR-130 suppresses adipogenesis by inhibiting peroxisome proliferator-activated receptor gamma expression. Mol Cell Biol. 31:626–638. 2011. View Article : Google Scholar
19	Pan S, Yang X, Jia Y, Li R and Zhao R: Microvesicle-shuttled miR-130b reduces fat deposition in recipient primary cultured porcine adipocytes by inhibiting PPAR-g expression. J Cell Physiol. 229:631–639. 2014. View Article : Google Scholar
20	Lewis BP, Burge CB and Bartel DP: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 120:15–20. 2005. View Article : Google Scholar : PubMed/NCBI
21	Zeng XC, Li XS and Wen H: Telmisartan protects against microvascular dysfunction during myocardial ischemia/reperfusion injury by activation of peroxisome proliferator-activated receptor γ. BMC Cardiovasc Disord. 13:392013. View Article : Google Scholar
22	Maass AH and Buvoli M: Cardiomyocyte preparation, culture and gene transfer. Methods Mol Biol. 366:321–330. 2007. View Article : Google Scholar
23	Krützfeldt J, Rajewsky N, Braich R, Rajeev KG, Tuschl T, Manoharan M and Stoffel M: Silencing of microRNAs in vivo with 'antagomirs'. Nature. 438:685–689. 2005. View Article : Google Scholar : PubMed/NCBI
24	Ren XP, Wu J, Wang X, Sartor MA, Qian J, Jones K, Nicolaou P, Pritchard TJ and Fan GC: MicroRNA-320 is involved in the regulation of cardiac ischemia/reperfusion injury by targeting heat-shock protein 20. Circulation. 119:2357–2366. 2009. View Article : Google Scholar : PubMed/NCBI
25	Herrera BM, Lockstone HE, Taylor JM, Wills QF, Kaisaki PJ, Barrett A, Camps C, Fernandez C, Ragoussis J, Gauguier D, et al: MicroRNA-125a is over-expressed in insulin target tissues in a spontaneous rat model of Type 2 diabetes. BMC Med Genomics. 2:542009. View Article : Google Scholar : PubMed/NCBI
26	Schmittgen TD and Livak KJ: Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 3:1101–1108. 2008. View Article : Google Scholar : PubMed/NCBI
27	Shan H, Li X, Pan Z, Zhang L, Cai B, Zhang Y, Xu C, Chu W, Qiao G, Li B, et al: Tanshinone IIA protects against sudden cardiac death induced by lethal arrhythmias via repression of microRNA-1. Br J Pharmacol. 158:1227–1235. 2009. View Article : Google Scholar : PubMed/NCBI
28	Zhong JQ, Zhang W, Gao H, Li Y, Zhong M, Li D, Zhang C and Zhang Y: Changes in connexin 43, metalloproteinase and tissue inhibitor of metalloproteinase during tachycardia-induced cardiomyopathy in dogs. Eur J Heart Fail. 9:23–29. 2007. View Article : Google Scholar
29	Li R, Yan G, Li Q, Sun H, Hu Y, Sun J and Xu B: MicroRNA-145 protects cardiomyocytes against hydrogen peroxide (H₂O₂)-induced apoptosis through targeting the mitochondria apoptotic pathway. PLoS One. 7:e449072012. View Article : Google Scholar
30	Wang X, Ha T, Liu L, Zou J, Zhang X, Kalbfleisch J, Gao X, Williams D and Li C: Increased expression of microRNA-146a decreases myocardial ischaemia/reperfusion injury. Cardiovasc Res. 97:432–442. 2013. View Article : Google Scholar :
31	Liu H, Guo X, Chu Y and Lu S: Heart protective effects and mechanism of quercetin preconditioning on anti-myocardial ischemia reperfusion (IR) injuries in rats. Gene. 545:149–155. 2014. View Article : Google Scholar : PubMed/NCBI
32	Ren-an Q, Juan L, Chuyuan L, Wenjuan F, Chunyan H, Xuemei Y, Lin H and Hong N: Study of the protective mechanisms of compound Danshen tablet (Fufang Danshen Pian) against myocardial ischemia/reperfusion injury via the Akt-eNOS signaling pathway in rats. J Ethnopharmacol. 156:190–198. 2014. View Article : Google Scholar : PubMed/NCBI
33	Fulton D, Gratton JP, McCabe TJ, Fontana J, Fujio Y, Walsh K, Franke TF, Papapetropoulos A and Sessa WC: Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature. 399:597–601. 1999. View Article : Google Scholar : PubMed/NCBI
34	Limaye V, Li X, Hahn C, Xia P, Berndt MC, Vadas MA and Gamble JR: Sphingosine kinase-1 enhances endothelial cell survival through a PECAM-1-dependent activation of PI-3K/Akt and regulation of Bcl-2 family members. Blood. 105:3169–3177. 2005. View Article : Google Scholar : PubMed/NCBI
35	Thomas RL, Roberts DJ, Kubli DA, Lee Y, Quinsay MN, Owens JB, Fischer KM, Sussman MA, Miyamoto S and Gustafsson ÅB: Loss of MCL-1 leads to impaired autophagy and rapid development of heart failure. Genes Dev. 27:1365–1377. 2013. View Article : Google Scholar : PubMed/NCBI
36	Wang X, Bathina M, Lynch J, Koss B, Calabrese C, Frase S, Schuetz JD, Rehg JE and Opferman JT: Deletion of MCL-1 causes lethal cardiac failure and mitochondrial dysfunction. Genes Dev. 27:1351–1364. 2013. View Article : Google Scholar : PubMed/NCBI