Pioglitazone alleviates oxygen and glucose deprivation-induced injury by up-regulation of miR-454 in H9c2 cells

Document Type: Original Article


1 Shandong University, Jinan 250100, Shangdong, China

2 Department of Cardiac Surgery, Linyi People’s Hospital, Linyi 276000, Shandong, China

3 Department of Equipment, Linyi People’s Hospital, Linyi 276000, Shandong, China

4 Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong, China


Objective(s): Pioglitazone, an anti-diabetic agent, has been widely used to treat type II diabetes. However, the effect of pioglitazone on myocardial ischemia reperfusion injury (MIRI) is still unclear. Herein, the objective of this study is to learn about the regulation and mechanism of pioglitazone effects on oxygen glucose deprivation (OGD)-induced myocardial cell injury.
Materials and Methods: A cellular injury model of OGD-treated H9c2 cells in vitro was constructed to simulate ischemic/reperfusion (I/R) injury. Then, various concentrations of pioglitazone (0, 2.5, 5, 7.5 and 10 μM) were used for the treatment of H9c2 cells, and CCK-8, flow cytometry and western blot assays were performed to examine cell viability, apoptosis, and the protein levels of factors involved in cell cycle and apoptosis in OGD-treated cells. MiR-454 inhibitor was used to suppress miR-454 expression, and whether miR-454 was involved in regulating OGD-induced cell injury was studied. Two key signal pathways were examined to uncover the underlying mechanism.
Results: OGD reduced cell proliferation and induced apoptosis in H9c2 cells (P<0.05, PConclusion: Pioglitazone protected H9c2 cells against OGD-induced injury through up-regulating miR-454, indicating a novel therapeutic strategy for treatment of MIRI.


Main Subjects

1. Bijkerk R, Van SC, de Boer HC, Van dPP, Khairoun M, de Bruin RG, et al. Hematopoietic microRNA-126 protects against renal ischemia/reperfusion injury by promoting vascular integrity. J Am Soc Nephrol 2014; 25:1710-1722.
2. Yang CJ, Yang J, Fan ZX, Yang J. Activating transcription factor 3 ‑ an endogenous inhibitor of myocardial ischemia-reperfusion injury (Review). Mol Med Rep 2015; 104:566-567.
3. Gonca E, Kurt Ç. Cardioprotective effect of thymoquinone: a constituent of Nigella sativa L., against myocardial ischemia/reperfusion injury and ventricular arrhythmias in anaesthetized rats. Pak J Pharm Sci 2015; 28:1267-1273.
4. Song CL, Liu B, Diao HY, Shi YF, Li YX, Zhang JC, et al. The protective effect of microRNA-320 on left ventricular remodeling after myocardial ischemia-reperfusion injury in the rat model. Int J Mol Sci 2014; 15:17442-17456.
5. Yu LN, Yu J, Zhang FJ, Yang MJ, Ding TT, Wang JK, et al. Sevoflurane postconditioning reduces myocardial reperfusion injury in rat isolated hearts via activation of PI3K/Akt signaling and modulation of Bcl-2 family proteins. J Zhejiang Univ Sci B 2010;11:661-672.
6. Xu M, Hao H, Jiang L, Wei Y, Zhou F, Sun J, et al. Cardiotonic pill reduces myocardial ischemia-reperfusion injury via increasing EET concentrations in rats. Drug Metab Dispos 2016; 44:878-887.
7. Jiang LY, Tang SS, Wang XY, Liu LP, Long Y, Hu M, et al. PPARγ agonist pioglitazone reverses memory impairment and biochemical changes in a mouse model of type 2 diabetes. CNS Neurosci Ther 2012; 18:659-666.
8. Doehner W, Anker SD. Pioglitazone after ischemic stroke or transient ischemic attack. New Engl J Med 2016; 64:1321-1331.
9. Heneka MT, Fink A, Doblhammer G. Effect of pioglitazone medication on the incidence of dementia. Ann Neurol 2015; 78:284–294.
10. Tuccori M, Filion KB, Hui Y, Yu OH, Platt RW, Azoulay L. Pioglitazone use and risk of bladder cancer: population based cohort study. Bmj 2016; 352:1541.
11. Yu L, Zhaodong J, Guan Y, Wang G, Wang Y. Protective effect of pioglitazone and insulin on cardiomyocyte after myocardial ischemia reperfusion injury in rats. Chin J Anat 2008; 31:658-661.
12. Tokutome M, Matoba T, Nakano Y, Nakano K, Sunagawa K, Egashira K. Nanoparticles-mediated delivery of pioglitazone reduces myocardial ischemia-reperfusion injury by antagonizing monocyte-mediated inflammation in mice and mini pigs. Am Heart Assoc 2014; 130:A17380.
13. Birnbaum Y, Ye Y, Lin Y, Freeberg SY, Nishi SP, Martinez JD, et al. Augmentation of myocardial production of 15-epi-lipoxin-a4 by pioglitazone and atorvastatin in the rat. Circulation 2006; 114:929-935.
14. Li J, Lang MJ, Mao XB, Tian L, Feng YB. Antiapoptosis and mitochondrial effect of pioglitazone preconditioning in the ischemic/reperfused heart of rat. Cardiovasc Drugs Ther 2008; 22:283-291.
15. Wang H, Zhu QW, Ye P, Li ZB, Li Y, Cao ZL, et al. Pioglitazone attenuates myocardial ischemia-reperfusion injury via up-regulation of ERK and COX-2. Biosci Trends 2012; 6:325-332.
16. Iaconetti C, Sorrentino S, De RS, Indolfi C. Exosomal miRNAs in Heart Disease. Physiology 2016; 31:16-24.
17. Dosenko VE, Gurianova VL, Surova OV, Stroy DA, Moibenko AA. Mature and immature microRNA ratios in cultured rat cardiomyocytes during anoxia-reoxygenation. Exp Clin Cardiol 2012; 17:84-87.
18. Chen F, Chen ZY, Yang HT. Expression profile of microRNAs in the cardiomyocytes derived from mouse embryonic stem cells. Sheng LI Xue Bao [Acta Physiologica Sinica] 2014; 66:702-708.
19. Lei Y, Gong X, Lei S, Hong Y, Lu B, Zhu L. miR-454 functions as an oncogene by inhibiting CHD5 in hepatocellular carcinoma. Oncotarget 2015; 6:39225-39234.
20. Tao Z, Yuan Y, Liao Q. Alleviation of lipopolysaccharides-induced acute lung injury by MiR-454. Cell Physiol Biochem 2016; 38:65-74.
21. Gad MZ, Ehssan NA, Ghiet MH, Wahman LF, Ghiet MH, Wahman LF. Effects of pioglitazone and metformin on carbohydrate metabolism in experimental models of glucose intolerance. Int J Diabetes Metab 2010; 18:132-138.
22. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2 −ΔΔ C T method. Methods 2012; 25:402-408.
23. Liu J, Yang S, Zhang X, Liu G, Yue X. Isoflurane reduces oxygen-glucose deprivation-induced oxidative, inflammatory, and apoptotic responses in H9c2 cardiomyocytes. Am J Transl Res 2016; 8:2597-2608.
24. Zheng K, Sheng Z, Li Y, Lu H. Salidroside inhibits oxygen glucose deprivation (OGD)/re-oxygenation-induced H9c2 cell necrosis through activating of Akt-Nrf2 signaling. Biochem Biophys Res Commun 2014; 451:79-85.
25. Wang XH. Construction of oxygen-glucose deprivation/reoxygenation injury model in the cultured neonatal rat myocytes. Ningxia Medical Journal 2012; 3:p:000.
26. Jintaek H, Han CK, Sangyoon C, Sungsoo K. Protective effect of dealcoholized persimmonwine on H2o2 - induced oxidative injury in H9c2 cardiomyocytes. J Food Res 2013; 2:61.
27. Ghosh S, Dey S. Pioglitazone induced weight changes in type 2 diabetic patients. Int J Collab Res Intern Med Public Health 2011; 3:534-540.
28. Honda T, Kaikita K, Tsujita K, Hayasaki T, Matsukawa M, Fuchigami S, et al. Pioglitazone, a peroxisome proliferator-activated receptor-gamma agonist, attenuates myocardial ischemia-reperfusion injury in mice with metabolic disorders. J Mol Cell Cardiol 2008; 44:915-926.
29. Ahmed LA, Salem HA, Attia AS, Agha AM. Pharmacological preconditioning with nicorandil and pioglitazone attenuates myocardial ischemia/reperfusion injury in rats. Eur J Pharmacol 2011; 663:51-58.
30. Liu P, Tian Y, Bao M, Wang Y, Cui F, Anesthiology DO. miRNA-214 was involved in cardioprotection by electroacupuncture pretreatment of myocardial ischemia / reperfusion injury in rats. Medical Journal of the Chinese Peoples Armed Police Force 2016.
31. Yang J, Chen L, Yang J, Ding J, Li S, Wu H, et al. MicroRNA-22 targeting CBP protects against myocardial ischemia-reperfusion injury through anti-apoptosis in rats. Mol Biol Rep 2014; 41:555-561.
32. Wang JX, Zhang XJ, Li Q, Wang K, Wang Y, Jiao JQ, et al. MicroRNA-103/107 regulate programmed necrosis and myocardial ischemia/reperfusion injury through targeting FADD. Circ Res 2015; 117:352-363.
33. Liang HL, Hu AP, Li SL, Xie JP, Ma QZ, Liu JY. MiR-454 prompts cell proliferation of human colorectal cancer cells by repressing CYLD expression. Asian Pac J Cancer Prev 2015; 16:2397-2402.
34. Du J, Tong A, Wang F, Cui Y, Li C, Zhang Y, et al. The roles of PI3K/AKT/mTOR and MAPK/ERK signaling pathways in human pheochromocytomas. Int J Endocrinol 2016; 2016:5286972.
35. Calvo N, Martín MJ, de Boland AR, Gentili C. Involvement of ERK1/2, p38 MAPK, and PI3K/Akt signaling pathways in the regulation of cell cycle progression by PTHrP in colon adenocarcinoma cells. Biochem Cell Biol 2014; 92:305-315.
36. Zhu YM, Wang CC, Chen L, Qian LB, Ma LL, Yu J, et al. Both PI3K/Akt and ERK1/2 pathways participate in the protection by dexmedetomidine against transient focal cerebral ischemia/reperfusion injury in rats. Brain Res 2013; 1494:1-8.
37. Thomas CJ, Lim NR, Kedikaetswe A, Yeap YY, Woodman OL, Ng DC, et al. Evidence that the MEK/ERK but not the PI3K/Akt pathway is required for protection from myocardial ischemia-reperfusion injury by 3’,4’-dihydroxyflavonol. Eur J Pharmacol 2015; 758:53-59.
38. Taniguchi Y, Ooie T, Takahashi N, Shinohara T, Nakagawa M, Yonemochi H, et al. Pioglitazone but not glibenclamide improves cardiac expression of heat shock protein 72 and tolerance against ischemia/reperfusion injury in the heredity insulin-resistant rat. Diabetes 2006; 55:2371-2378.
39. Zhao YQ, Zhao-Dong J, Guan YJ. Research of the mechanism of pioglitazone on myocardial ischemic reperfusion in jury by PI3K and ERK1/2 signaling pathway. Chin J Lab Diagnosis 2012; 2:p:007.