Role of miR-1 and miR-133a in myocardial ischemic postconditioning
Tóm tắt
Ischemic postconditioning (IPost) has aroused much attention since 2003 when it was firstly reported. The role of microRNAs (miRNAs or miRs) in IPost has rarely been reported. The present study was undertaken to investigate whether miRNAs were involved in the protective effect of IPost against myocardial ischemia-reperfusion (IR) injury and the probable mechanisms involved.
Thirty SD rats weighing 250-300 g were equally randomized to three groups: Control group, where the rats were treated with thoracotomy only; IR group, where the rats were treated with ischemia for 60 min and reperfusion for 180 min; and IPost group, where the rats were treated with 3 cycles of transient IR just before reperfusion. The extent of myocardial infarction, LDH and CK activities were measured immediately after treatment. Myocardial apoptosis was detected by TUNEL assay. The myocardial tissue was collected after IR or IPost stimulation to evaluate the miRNAs expression level by miRNA-microarray and quantitative real-time RT-PCR. Real-time PCR was conducted to identify changes in mRNA expression of apoptosis-related genes such as Bcl-2, Bax and Caspase-9 (CASP9), and Western blot was used to compare the protein expression level of CASP9 in the three groups. The miRNA mimics and anti-miRNA oligonucleotides (AMO) were transferred into the cultured neonatal cardiomyocytes and myocardium before they were treated with IR. The effect of miRNAs on apoptosis was determined by flow cytometry and TUNEL assay. CASP9, as one of the candidate target of miR-133a, was compared during IR after the miR-133a mimic or AMO-133a was transferred into the myocardium.
IPost reduced the IR-induced infarct size of the left ventricle, and decreased CK and LDH levels. TUNEL assay showed that myocardial apoptosis was attenuated by IPost compared with IR. MiRNA-microarray and RT-PCR showed that myocardial-specific miR-1 and miR-133a were down-regulated by IR, and up-regulated by IPost compared with IR. Furthermore, IPost up-regulated the mRNA expression of Bcl-2, down-regulated that of Bax and CASP9. Western blot showed that IPost also down-regulated the CASP9 protein expression compared with IR. The results of flow cytometry and TUNEL assay showed that up-regulation of miR-1 and miR-133a decreased apoptosis of cardiomyocytes. MiR-133a mimic down-regulated CASP9 protein expression and attenuated IR-induced apoptosis.
MiRNAs are associated with the protective effect of IPost against myocardial IR injury. IPost can up-regulate miR-1 and miR-133a, and decrease apoptosis of cardiomyocyte. Myocardial-specific miR-1 and miR-133a may play an important role in IPost protection by regulating apoptosis-related genes. MiR-133a may attenuate apoptosis of myocardiocytes by targeting CASP9.
Từ khóa
Tài liệu tham khảo
Cokkinos DV, Pantos C: Myocardial protection in man--from research concept to clinical practice. Heart Fail Rev. 2007, 12: 345-362. 10.1007/s10741-007-9030-5.
Wu HH, Hsiao TY, Chien CT, Lai MK: Ischemic conditioning by short periods of reperfusion attenuates renal ischemia/reperfusion induced apoptosis and autophagy in the rat. J Biomed Sci. 2009, 16: 19-10.1186/1423-0127-16-19.
Murry CE, Jennings RB, Reimer KA: Preconditioning with ischemia: A delay of lethal cell injury in ischemic myocardium. Circulation. 1986, 74: 1124-1136.
Zhao ZQ, Corvera JS, Halkos ME, Kerendi F, Wang NP, Guyton RA, Vinten-Johansen J: Inhibition of myocardial injury by ischemic postconditioning during reperfusion: Comparison with ischemic preconditioning. Am J Physiol Heart Circ Physiol. 2003, 285: H579-H588.
Kin H, Zhao ZQ, Sun HY, Wang NP, Corvera JS, Halkos ME, Kerendi F, Guyton RA, Vinten-Johansen J: Postconditioning attenuates myocardial ischemia-reperfusion injury by inhibiting events in the early minutes of reperfusion. Cardiovasc Res. 2004, 62: 74-85. 10.1016/j.cardiores.2004.01.006.
Argaud L, Gateau-Roesch O, Raisky O, Loufouat J, Robert D, Ovize M: Postconditioning inhibits mitochondrial permeability transition. Circulation. 2005, 111: 194-197. 10.1161/01.CIR.0000151290.04952.3B.
Thibault H, Piot C, Staat P, Bontemps L, Sportouch C, Rioufol G, Cung TT, Bonnefoy E, Angoulvant D, Aupetit JF, Finet G, Andre-Fouet X, Macia JC, Raczka F, Rossi R, Itti R, Kirkorian G, Derumeaux G, Ovize M: Long-term benefit of postconditioning. Circulation. 2008, 117: 1037-1044. 10.1161/CIRCULATIONAHA.107.729780.
Vinten-Johansen J: Postconditioning: A mechanical maneuver that triggers biological and molecular cardioprotective responses to reperfusion. Heart Fail Rev. 2007, 12: 235-244. 10.1007/s10741-007-9024-3.
Lin XM, Zhang ZY, Wang LF, Zhang L, Liu Y, Liu XL, Yang XC, Cui L: Attenuation of tumor necrosis factor-alpha elevation and improved heart function by postconditioning for 60 seconds in patients with acute myocardial infarction. Chin Med J (Engl). 2010, 123: 1833-1839.
Li Y, Ge X, Liu X: The cardioprotective effect of postconditioning is mediated by ARC through inhibiting mitochondrial apoptotic pathway. Apoptosis. 2009, 14: 164-172. 10.1007/s10495-008-0296-4.
Fang J, Wu L, Chen L: Postconditioning attenuates cardiocyte ultrastructure injury and apoptosis by blocking mitochondrial permeability transition in rats. Acta Cardiol. 2008, 63: 377-387. 10.2143/AC.63.3.1020316.
Penna C, Perrelli MG, Raimondo S, Tullio F, Merlino A, Moro F, Geuna S, Mancardi D, Pagliaro P: Postconditioning induces an anti-apoptotic effect and preserves mitochondrial integrity in isolated rat hearts. Biochim Biophys Acta. 2009, 1787: 794-801. 10.1016/j.bbabio.2009.03.013.
Zhao WS, Xu L, Wang LF, Zhang L, Zhang ZY, Liu Y, Liu XL, Yang XC, Cui L: A 60-s postconditioning protocol by percutaneous coronary intervention inhibits myocardial apoptosis in patients with acute myocardial infarction. Apoptosis. 2009, 14: 1204-1211. 10.1007/s10495-009-0387-x.
Bostjancic E, Zidar N, Stajer D, Glavac D: Micrornas mir-1, mir-133a, mir-133b and mir-208 are dysregulated in human myocardial infarction. Cardiology. 2010, 115: 163-169. 10.1159/000268088.
Xu C, Lu Y, Pan Z, Chu W, Luo X, Lin H, Xiao J, Shan H, Wang Z, Yang B: The muscle-specific micrornas mir-1 and mir-133 produce opposing effects on apoptosis by targeting hsp60, hsp70 and caspase-9 in cardiomyocytes. J Cell Sci. 2007, 120: 3045-3052. 10.1242/jcs.010728.
Ren XP, Wu J, Wang X, Sartor MA, Qian J, Jones K, Nicolaou P, Pritchard TJ, Fan GC: Microrna-320 is involved in the regulation of cardiac ischemia/reperfusion injury by targeting heat-shock protein 20. Circulation. 2009, 119: 2357-2366. 10.1161/CIRCULATIONAHA.108.814145.
Rane S, He M, Sayed D, Vashistha H, Malhotra A, Sadoshima J, Vatner DE, Vatner SF, Abdellatif M: Downregulation of mir-199a derepresses hypoxia-inducible factor-1alpha and sirtuin 1 and recapitulates hypoxia preconditioning in cardiac myocytes. Circ Res. 2009, 104: 879-886. 10.1161/CIRCRESAHA.108.193102.
Zhuo Y, Chen PF, Zhang AZ, Zhong H, Chen CQ, Zhu YZ: Cardioprotective effect of hydrogen sulfide in ischemic reperfusion experimental rats and its influence on expression of survivin gene. Biol Pharm Bull. 2009, 32: 1406-1410. 10.1248/bpb.32.1406.
Sodha NR, Clements RT, Feng J, Liu Y, Bianchi C, Horvath EM, Szabo C, Sellke FW: The effects of therapeutic sulfide on myocardial apoptosis in response to ischemia-reperfusion injury. Eur J Cardiothorac Surg. 2008, 33: 906-913. 10.1016/j.ejcts.2008.01.047.
Lee AS, Su MJ: Comparison of the cardiac effects between quinazoline-based alpha1-adrenoceptor antagonists on occlusion-reperfusion injury. J Biomed Sci. 2008, 15: 239-249. 10.1007/s11373-007-9214-y.
Shyu KG, Liang YJ, Chang H, Wang BW, Leu JG, Kuan P: Enhanced expression of angiopoietin-2 and the Tie2 receptor but not angiopoietin-1 or the Tie1 receptor in a rat model of myocardial infarction. J Biomed Sci. 2004, 11: 163-171. 10.1007/BF02256559.
Yin C, Salloum FN, Kukreja RC: A novel role of microrna in late preconditioning: Upregulation of endothelial nitric oxide synthase and heat shock protein 70. Circ Res. 2009, 104: 572-575. 10.1161/CIRCRESAHA.108.193250.
Sadoshima J, Jahn L, Takahashi T, Kulik TJ, Izumo S: Molecular characterization of the stretch-induced adaptation of cultured cardiac cells. An in vitro model of load-induced cardiac hypertrophy. J Biol Chem. 1992, 267: 10551-10560.
Zhou X, Sheng Y, Yang R, Kong X: Nicotine promotes cardiomyocyte apoptosis via oxidative stress and altered apoptosis-related gene expression. Cardiology. 2010, 115: 243-250. 10.1159/000301278.
Lopez-Neblina F, Toledo AH, Toledo-Pereyra LH: Molecular biology of apoptosis in ischemia and reperfusion. J Invest Surg. 2005, 18: 335-350. 10.1080/08941930500328862.
Kumar P, Coltas IK, Kumar B, Chepeha DB, Bradford CR, Polverini PJ: Bcl-2 protects endothelial cells against gamma-radiation via a raf-mek-erk-survivin signaling pathway that is independent of cytochrome c release. Cancer Res. 2007, 67: 1193-1202. 10.1158/0008-5472.CAN-06-2265.
Lalier L, Cartron PF, Juin P, Nedelkina S, Manon S, Bechinger B, Vallette FM: Bax activation and mitochondrial insertion during apoptosis. Apoptosis. 2007, 12: 887-896. 10.1007/s10495-007-0749-1.
Ikeda S, Kong SW, Lu J, Bisping E, Zhang H, Allen PD, Golub TR, Pieske B, Pu WT: Altered microrna expression in human heart disease. Physiol Genomics. 2007, 31: 367-373. 10.1152/physiolgenomics.00144.2007.
Salloum FN, Yin C, Kukreja RC: Role of miRs in Cardiac Preconditioning. J Cardiovasc Pharmacol. 2010,
Yin C, Wang X, Kukreja RC: Endogenous micrornas induced by heat-shock reduce myocardial infarction following ischemia-reperfusion in mice. FEBS Lett. 2008, 582: 4137-4142. 10.1016/j.febslet.2008.11.014.
Bostjancic E, Zidar N, Glavac D: Microrna microarray expression profiling in human myocardial infarction. Dis Markers. 2009, 27: 255-268.
Liu N, Williams AH, Kim Y, McAnally J, Bezprozvannaya S, Sutherland LB, Richardson JA, Bassel-Duby R, Olson EN: An intragenic mef2-dependent enhancer directs muscle-specific expression of micrornas 1 and 133. Proc Natl Acad Sci USA. 2007, 104: 20844-20849. 10.1073/pnas.0710558105.
Horie T, Ono K, Nishi H, Iwanaga Y, Nagao K, Kinoshita M, Kuwabara Y, Takanabe R, Hasegawa K, Kita T, Kimura T: Microrna-133 regulates the expression of glut4 by targeting klf15 and is involved in metabolic control in cardiac myocytes. Biochem Biophys Res Commun. 2009, 389: 315-320. 10.1016/j.bbrc.2009.08.136.
Luo X, Lin H, Pan Z, Xiao J, Zhang Y, Lu Y, Yang B, Wang Z: Down-regulation of mir-1/mir-133 contributes to re-expression of pacemaker channel genes hcn2 and hcn4 in hypertrophic heart. J Biol Chem. 2008, 283: 20045-20052. 10.1074/jbc.M801035200.