Abstract
Stroke can cause death and disability and has a high incidence with many complications. So far, effective treatment options for stroke are still limited. MicroRNA-532-5p (miR-532-5p) is significantly downregulated in stroke. However, the role of miR-532-5p in ischemic stroke is still unclear. In this study, we established an in vivo middle cerebral artery occlusion (MCAO) model in mice. The expression level of miR-532-5p, neurological score, infarct area, neuronal apoptosis, and phosphoinositide 3-kinase (PI3K)/Akt signaling pathway-related molecules were examined. Low miR-532-5p levels and high phosphatase and tensin homolog deleted on chromosome 10 (PTEN) levels were detected in the mouse MCAO model. MiR-532-5p overexpression improved neurological dysfunction, reduced the infarct area, attenuated neuronal injury and apoptosis, and promoted the activation of the PI3K/Akt signaling pathway in MCAO mice. In vitro, we treated mouse neuroblastoma cells (N2a) with oxygen-glucose deprivation and reperfusion (OGD/R). The expression level of miR-532-5p, cell viability, cell apoptosis, and the PI3K/Akt signaling pathway-related molecules were detected. Consistent with the in vivo tests, the miR-532-5p level was decreased and the PTEN level was increased in OGD-treated N2a cells in vitro. The miR-532-5p mimic increased cell viability, decreased cell apoptosis, and activated the PI3K/Akt signaling pathway. Furthermore, PTEN was verified as a target gene of miR-532-5p by luciferase reporter assay. PTEN overexpression attenuated the protective effect of miR-532-5p in OGD-treated N2a cells. In summary, these findings reveal that miR-532-5p protects against ischemic stroke by inhibiting PTEN and activating the PI3K/Akt signaling pathway and may serve as a novel therapeutic target for ischemic stroke.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adelson JD et al (2012) Neuroprotection from stroke in the absence of MHCI or PirB. Neuron 73:1100–1107. https://doi.org/10.1016/j.neuron.2012.01.020
Barca-Mayo O, De Pietri Tonelli D (2014) Convergent microRNA actions coordinate neocortical development. Cell Mol Life Sci: CMLS 71:2975–2995. https://doi.org/10.1007/s00018-014-1576-5
Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136:215–233. https://doi.org/10.1016/j.cell.2009.01.002
Bayoumi AS et al (2017) MicroRNA-532 protects the heart in acute myocardial infarction, and represses prss23, a positive regulator of endothelial-to-mesenchymal transition. Cardiovasc Res 113:1603–1614. https://doi.org/10.1093/cvr/cvx132
Beal CC (2010) Gender and stroke symptoms: a review of the current literature. J Neurosci Nurs: Journal of the American Association of Neuroscience Nurses 42:80–87
Bhuiyan MIH, Jung SY, Kim HJ, Lee YS, Jin C (2011) Major role of the PI3K/Akt pathway in ischemic tolerance induced by sublethal oxygen-glucose deprivation in cortical neurons in vitro. Arch Pharm Res 34:1023–1034. https://doi.org/10.1007/s12272-011-0620-3
Bhalala OG, Srikanth M, Kessler JA (2013) The emerging roles of microRNAs in CNS injuries. Nat Rev Neurol 9:328-339. https://doi.org/10.1038/nrneurol.2013.67
Chen P-H, Gao S, Wang Y-J, Xu A-D, Li Y-S, Wang D (2012) Classifying ischemic stroke, from TOAST to CISS. CNS Neurosci Ther 18:452–456. https://doi.org/10.1111/j.1755-5949.2011.00292.x
Cheng X, Ander BP, Jickling GC, Zhan X, Hull H, Sharp FR, Stamova B (2019) MicroRNA and their target mRNAs change expression in whole blood of patients after intracerebral hemorrhage. J Cereb Blood Flow Metab. https://doi.org/10.1177/0271678x19839501
Delgado-Esteban M, Martin-Zanca D, Andres-Martin L, Almeida A, Bolaños JP (2007) Inhibition of PTEN by peroxynitrite activates the phosphoinositide-3-kinase/Akt neuroprotective signaling pathway. J Neurochem 102:194–205. https://doi.org/10.1111/j.1471-4159.2007.04450.x
Doench JG, Sharp PA (2004) Specificity of microRNA target selection in translational repression. Genes Dev 18:504–511. https://doi.org/10.1101/gad.1184404
Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35:495–516. https://doi.org/10.1080/01926230701320337
Eyileten C et al. (2018) MicroRNAs as Diagnostic and Prognostic Biomarkers in Ischemic Stroke-A Comprehensive Review and Bioinformatic Analysis. Cells 7, 249. https://doi.org/10.3390/cells7120249
Garcia JH, Wagner S, Liu KF, Hu XJ (1995) Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Statistical validation Stroke 26:627–634; discussion 635 https://doi.org/10.1161/01.str.26.4.627
Griesing S et al (2017) Thyroid transcription factor-1-regulated microRNA-532-5p targets KRAS and MKL2 oncogenes and induces apoptosis in lung adenocarcinoma cells. Cancer Sci 108:1394–1404. https://doi.org/10.1111/cas.13271
Helmschrodt C et al. (2017) Polyethylenimine Nanoparticle-Mediated siRNA Delivery to Reduce α-Synuclein Expression in a Model of Parkinson’s Disease Molecular Therapy - Nucleic Acids 9:57–68 https://doi.org/10.1016/j.omtn.2017.08.013
Hirai K et al (2004) PI3K inhibition in neonatal rat brain slices during and after hypoxia reduces phospho-Akt and increases cytosolic cytochrome c and apoptosis. Brain Res Mol Brain Res 124:51–61. https://doi.org/10.1016/j.molbrainres.2004.02.009
Hu L, Sun Y, Hu J (2010) Catalpol inhibits apoptosis in hydrogen peroxide-induced endothelium by activating the PI3K/Akt signaling pathway and modulating expression of Bcl-2 and Bax. Eur J Pharmacol 628:155–163. https://doi.org/10.1016/j.ejphar.2009.11.046
Jiang M et al (2014) Neuroprotective effects of bilobalide on cerebral ischemia and reperfusion injury are associated with inhibition of pro-inflammatory mediator production and down-regulation of JNK1/2 and p38 MAPK activation. J Neuroinflammation 11:167. https://doi.org/10.1186/s12974-014-0167-6
John B, Enright AJ, Aravin A, Tuschl T, Sander C, Marks DS (2004) Human MicroRNA targets. PLoS Biol 2:e363. https://doi.org/10.1371/journal.pbio.0020363
Khare D, Goldschmidt N, Bardugo A, Gur-Wahnon D, Ben-Dov IZ, Avni B (2017) Plasma microRNA profiling: Exploring better biomarkers for lymphoma surveillance. PLoS One 12:e0187722. https://doi.org/10.1371/journal.pone.0187722
Kilic U et al (2017) Particular phosphorylation of PI3K/Akt on Thr308 via PDK-1 and PTEN mediates melatonin's neuroprotective activity after focal cerebral ischemia in mice. Redox Biol 12:657–665. https://doi.org/10.1016/j.redox.2017.04.006
Krishnamurthi RV et al (2013) Global and regional burden of first-ever ischaemic and haemorrhagic stroke during 1990-2010: findings from the Global Burden of Disease Study 2010. Lancet Glob Health 1:e259–e281. https://doi.org/10.1016/S2214-109X(13)70089-5
Li D, Qu Y, Mao M, Zhang X, Li J, Ferriero D, Mu D (2009) Involvement of the PTEN-AKT-FOXO3a pathway in neuronal apoptosis in developing rat brain after hypoxia-ischemia. J Cerebr Blood Flow Metab: Official Journal of the International Society of Cerebral Blood Flow and Metabolism 29:1903–1913. https://doi.org/10.1038/jcbfm.2009.102
Li P, Teng F, Gao F, Zhang M, Wu J, Zhang C (2015) Identification of circulating microRNAs as potential biomarkers for detecting acute ischemic stroke. Cell Mol Neurobiol 35:433–447. https://doi.org/10.1007/s10571-014-0139-5
Liguori M et al (2018) Dysregulation of MicroRNAs and target genes networks in peripheral blood of patients with sporadic amyotrophic lateral sclerosis. Front Mol Neurosci 11:288. https://doi.org/10.3389/fnmol.2018.00288
Liu B et al (2010a) Preservation of GABAA receptor function by PTEN inhibition protects against neuronal death in ischemic stroke. Stroke 41:1018–1026. https://doi.org/10.1161/STROKEAHA.110.579011
Liu C, Wu J, Xu K, Cai F, Gu J, Ma L, Chen J (2010b) Neuroprotection by baicalein in ischemic brain injury involves PTEN/AKT pathway. J Neurochem 112:1500–1512. https://doi.org/10.1111/j.1471-4159.2009.06561.x
Liu DZ et al (2016) Elevating microRNA-122 in blood improves outcomes after temporary middle cerebral artery occlusion in rats. J Cerebr Blood Flow Metab: Official Journal of the International Society of Cerebral Blood Flow and Metabolism 36:1374–1383. https://doi.org/10.1177/0271678X15610786
Luo HR et al (2003) Akt as a mediator of cell death. Proc Natl Acad Sci U S A 100:11712–11717. https://doi.org/10.1073/pnas.1634990100
Ma J et al (2018) MiR-532-5p alleviates hypoxia-induced cardiomyocyte apoptosis by targeting PDCD4. Gene 675:36–43. https://doi.org/10.1016/j.gene.2018.06.087
Maehama T, Dixon JE (1998) The tumor suppressor, PTEN/MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3,4,5-trisphosphate. J Biol Chem 273:13375–13378. https://doi.org/10.1074/jbc.273.22.13375
Mokin M, Rojas H, Levy EI (2016) Randomized trials of endovascular therapy for stroke — impact on stroke care. Nat Rev Neurol 12:86–94. https://doi.org/10.1038/nrneurol.2015.240
Nampoothiri SS, Rajanikant GK (2019) miR-9 Upregulation integrates post-ischemic neuronal survival and regeneration in vitro. Cell Mol Neurobiol 39:223–240. https://doi.org/10.1007/s10571-018-0642-1
O'Brien J, Hayder H, Zayed Y, Peng C (2018) Overview of MicroRNA biogenesis, mechanisms of actions, and circulation. Front Endocrinol 9:402. https://doi.org/10.3389/fendo.2018.00402
Pan J-J et al (2011) Activation of Akt/GSK3β and Akt/Bcl-2 signaling pathways in nickel-transformed BEAS-2B cells. Int J Oncol 39:1285–1294. https://doi.org/10.3892/ijo.2011.1157
Qiu J et al (2016) Sinomenine activates astrocytic dopamine D2 receptors and alleviates neuroinflammatory injury via the CRYAB/STAT3 pathway after ischemic stroke in mice. J Neuroinflammation 13:263. https://doi.org/10.1186/s12974-016-0739-8
Rink C, Khanna S (2011) MicroRNA in ischemic stroke etiology and pathology. Physiol Genomics 43:521–528. https://doi.org/10.1152/physiolgenomics.00158.2010
Selmaj I, Cichalewska M, Namiecinska M, Galazka G, Horzelski W, Selmaj KW, Mycko MP (2017) Global exosome transcriptome profiling reveals biomarkers for multiple sclerosis. Ann Neurol 81:703–717. https://doi.org/10.1002/ana.24931
Sepramaniam S et al (2014) Circulating microRNAs as biomarkers of acute stroke. Int J Mol Sci 15:1418–1432. https://doi.org/10.3390/ijms15011418
Shi GD, OuYang YP, Shi JG, Liu Y, Yuan W, Jia LS (2011) PTEN deletion prevents ischemic brain injury by activating the mTOR signaling pathway. Biochem Biophys Res Commun 404:941–945. https://doi.org/10.1016/j.bbrc.2010.12.085
Song Y, Zhao Y, Ding X, Wang X (2018) microRNA-532 suppresses the PI3K/Akt signaling pathway to inhibit colorectal cancer progression by directly targeting IGF-1R. Am J Cancer Res 8:435–449
Stambolic V et al (1998) Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell 95:29–39. https://doi.org/10.1016/s0092-8674(00)81780-8
Talebi A, Rahnema M, Bigdeli MR (2019) Effect of intravenous injection of antagomiR-1 on brain ischemia. Mol Biol Rep 46:1149–1155. https://doi.org/10.1007/s11033-018-04580-y
Tan JR, Tan KS, Koo YX, Yong FL, Wang CW, Armugam A, Jeyaseelan K (2013) Blood microRNAs in low or no risk ischemic stroke patients. Int J Mol Sci 14:2072–2084. https://doi.org/10.3390/ijms14012072
Waite KA, Eng C (2002) Protean PTEN: form and function. Am J Hum Genet 70:829–844. https://doi.org/10.1086/340026
Wang F, Chang JT-H, Kao CJ, Huang RS (2016) High expression of miR-532-5p, a tumor suppressor, leads to better prognosis in ovarian cancer both in vivo and in vitro. Mol Cancer Ther 15:1123–1131. https://doi.org/10.1158/1535-7163.MCT-15-0943
Wei H, Tang Q-L, Zhang K, Sun J-J, Ding R-F (2018) miR-532-5p is a prognostic marker and suppresses cells proliferation and invasion by targeting TWIST1 in epithelial ovarian cancer. Eur Rev Med Pharmacol Sci 22:5842–5850. https://doi.org/10.26355/eurrev_201809_15911
Xu X, Zhang Y, Liu Z, Zhang X, Jia J (2016) miRNA-532-5p functions as an oncogenic microRNA in human gastric cancer by directly targeting RUNX3. J Cell Mol Med 20:95–103. https://doi.org/10.1111/jcmm.12706
Yang GY, Zhao YJ, Davidson BL, Betz AL (1997) Overexpression of interleukin-1 receptor antagonist in the mouse brain reduces ischemic brain injury. Brain Res 751:181–188. https://doi.org/10.1016/s0006-8993(96)01277-2
Yin K-J, Deng Z, Huang H, Hamblin M, Xie C, Zhang J, Chen YE (2010) miR-497 regulates neuronal death in mouse brain after transient focal cerebral ischemia. Neurobiol Dis 38:17–26. https://doi.org/10.1016/j.nbd.2009.12.021
Yu Z-H et al (2016) PI3K/Akt pathway contributes to neuroprotective effect of Tongxinluo against focal cerebral ischemia and reperfusion injury in rats. J Ethnopharmacol 181:8–19. https://doi.org/10.1016/j.jep.2016.01.028
Zhang L, Qu Y, Tang J, Chen D, Fu X, Mao M, Mu D (2010) PI3K/Akt signaling pathway is required for neuroprotection of thalidomide on hypoxic-ischemic cortical neurons in vitro. Brain Res 1357:157–165. https://doi.org/10.1016/j.brainres.2010.08.007
Zhang R et al (2019) miR-1247-3p mediates apoptosis of cerebral neurons by targeting caspase-2 in stroke. Brain Res 1714:18–26. https://doi.org/10.1016/j.brainres.2019.02.020
Zhao H, Sapolsky RM, Steinberg GK (2006) Phosphoinositide-3-kinase/akt survival signal pathways are implicated in neuronal survival after stroke. Mol Neurobiol 34:249–270. https://doi.org/10.1385/MN:34:3:249
Zuo W, Yan F, Zhang B, Hu X, Mei D (2018) Salidroside improves brain ischemic injury by activating PI3K/Akt pathway and reduces complications induced by delayed tPA treatment. Eur J Pharmacol 830:128–138. https://doi.org/10.1016/j.ejphar.2018.04.001
Acknowledgements
This work was supported by the Key research and development projects of Liaoning province, China.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
There are no conflicts of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Mu, J., Cheng, X., Zhong, S. et al. Neuroprotective effects of miR-532-5p against ischemic stroke. Metab Brain Dis 35, 753–763 (2020). https://doi.org/10.1007/s11011-020-00544-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11011-020-00544-z