Sheng Li, Fang-Yi Xiao, Pei-Ren Shan, Lan Su, De-Liang Chen, Jin-Ye Ding, Zhi-Quan Wang
To examine microRNA-133a (miR-133a) endogenous expression in cardiomyocytes after ischemia-reperfusion (I/R) injury and study the effects of miR-133a overexpression on I/R injury-induced cardiomyocyte apoptosis. Dual-Luciferase Reporter Assay detected dynamic expression of miR-133a. In an in vitro hypoxia-reoxygenation (HR) injury model and an in vivo rat model of I/R injury, rat cardiomyocytes were transfected with miR-133a mimic to test the effects of miR-133a overexpression on apoptosis. MiR-133a and Death Associated Protein Kinase 2 (DAPK2) mRNA expression was measured using real-time-PCR, and DAPK2 protein expression was detected by western blotting. Annexin V-fluorescein isothiocyanate/propidium iodide (PI) double-staining measured the apoptosis rate in H9C2 cells and transferase dUTP nick end labeling assay quantified the cardiomyocyte apoptosis rate in tissues obtained from in vivo the rat model. DAPK2 is a target of miR-133a. Both in vitro and in vivo results confirmed that after expression of miR-133a mimics, miR-133a levels increased, which was accompanied by decrease in DAPK2 mRNA and protein expression. In H9C2 cells, HR injury caused a sharp decrease in miR-133a expression and a significant upregualtion of DAPK2 mRNA and protein levels. However, exogenous miR-133a expression led to a significant reduction in DAPK2 mRNA and protein levels despite HR injury. Similar results were obtained from in vivo I/R injury model. After HR injury or I/R injury the apoptosis rate of myocardial cells was highly elevated and decreased significantly only after transfection of miR-133a into cardiomyocytes. MiR-133a overexpression may inhibit I/R injury-mediated cardiomyocyte apoptosis by targeting DAPK2, leading to reduced DAPK2 protein, thus miR-133a may potentially have a high therapeutic value in I/R injury.
J Mol Cell Cardiol. 2007 Jun;42(6):1137-41
[PMID:
17498736]
Hypertension. 2010 Apr;55(4):946-52
[PMID:
20177001]
J Mol Cell Cardiol. 2012 Apr;52(4):822-31
[PMID:
21889943]
Physiol Genomics. 2011 May 1;43(10):534-42
[PMID:
20959496]
Semin Cardiothorac Vasc Anesth. 2012 Sep;16(3):123-32
[PMID:
22368166]
Cell Stem Cell. 2008 Mar 6;2(3):219-29
[PMID:
18371447]
Methods. 2001 Dec;25(4):402-8
[PMID:
11846609]
J Biol Chem. 2008 Jul 18;283(29):20045-52
[PMID:
18458081]
Stem Cell Reports. 2014 Dec 9;3(6):1029-42
[PMID:
25465869]
Int J Biochem Cell Biol. 2010 Aug;42(8):1252-5
[PMID:
20619221]
PLoS One. 2013 Apr 29;8(4):e62400
[PMID:
23638067]
Cell Death Dis. 2014 Feb 27;5:e1082
[PMID:
24577080]
Circ Res. 2009 Jan 30;104(2):170-8, 6p following 178
[PMID:
19096030]
Circ Cardiovasc Genet. 2011 Aug 1;4(4):446-54
[PMID:
21642241]
Genes Dev. 2008 Dec 1;22(23):3242-54
[PMID:
19015276]
J Cell Sci. 2011 Sep 15;124(Pt 18):3187
[PMID:
21914820]
J Cell Sci. 2007 Sep 1;120(Pt 17):3045-52
[PMID:
17715156]
Curr Pharm Biotechnol. 2014;15(5):430-7
[PMID:
24846068]
PLoS Med. 2006 Nov;3(11):e442
[PMID:
17132052]
FEBS J. 2010 Jan;277(1):48-57
[PMID:
19878313]
Circ Res. 2007 Feb 16;100(3):416-24
[PMID:
17234972]
Oncogene. 2008 Sep 25;27(43):5706-16
[PMID:
18521079]
Nat Rev Cardiol. 2011 Jun 21;8(11):619-29
[PMID:
21691310]
J Biomed Sci. 2011 Mar 16;18:22
[PMID:
21406115]
Circulation. 2009 May 5;119(17):2357-66
[PMID:
19380620]
Circ Res. 2010 Sep 3;107(5):677-84
[PMID:
20595655]
Cardiovasc Res. 2010 Aug 1;87(3):431-9
[PMID:
20219857]
Nat Med. 2007 May;13(5):613-8
[PMID:
17468766]
Cardiovasc Res. 2012 May 1;94(2):284-92
[PMID:
22038740]
J Biol Chem. 2007 Apr 27;282(17):12363-7
[PMID:
17344217]
J Cardiovasc Pharmacol Ther. 2015 Mar;20(2):211-20
[PMID:
24924917]
J Mol Neurosci. 2014 Jun;53(2):242-50
[PMID:
24696166]
Physiol Rev. 2012 Jul;92 (3):1393-478
[PMID:
22988594]
J Clin Invest. 2013 Jan;123(1):92-100
[PMID:
23281415]
Eur Heart J. 2013 Jun;34(23):1714-22
[PMID:
23536610]
Sci Transl Med. 2014 Jun 4;6(239):239ps3
[PMID:
24898744]
