Abstract
This study aimed to establish the role of miR-129 and miR-384-5p in cerebral ischemia-induced apoptosis. Using PC12 cells transfected with miR-129 or miR-384-5p mimics or inhibitors, oxygen glucose deprivation (OGD) conditions were applied for 4 h to simulate transient cerebral ischemia. Apoptotic phenotypes were assessed via lactate dehydrogenase (LDH) assay, MTT cell metabolism assay, and fluorescence-activated cell sorting (FACS). The effect of miR overexpression and inhibition was evaluated by protein and mRNA detection of bcl-2 and caspase-3, critical apoptosis factors. Finally, the direct relationship of miR-129 and bcl-2 and miR-384-5p and caspase-3 was measured by luciferase reporter assay. The overexpression of miR-384-5p and miR-129 deficiency significantly enhanced cell viability, reduced LDH release, and inhibited apoptosis. By contrast, overexpression of miR-129 and miR-384-5p deficiency aggravated hypoxia-induced apoptosis and cell injury. miR-129 overexpression significantly reduced mRNA and protein levels of bcl-2 and miR-129 inhibition significantly increased mRNA and protein levels of bcl-2 in hypoxic cells.miR-384-5p overexpression significantly reduced protein levels of caspase-3 while miR-384-5p deficiency significantly increased protein levels of caspase-3. However, no changes were observed in caspase-3 mRNA in either transfection paradigm. Finally, luciferase reporter assay confirmed caspase-3 to be a direct target of miR-384-5p; however, no binding activity was detected between bcl-2 and miR-129.Transient cerebral ischemia induces differential expression of miR-129 and miR-384-5p which influences apoptosis by regulating apoptotic factors caspase-3 and bcl-2, thereby participating in the pathological mechanism of cerebral ischemia, and becoming potential targets for the treatment of ischemic cerebral injury in the future.
Graphic abstract
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Availability of data and materials
The dataset(s) supporting the conclusions of this article are available from the corresponding author upon the reasonable request.
References
Akpan N, Troy CM (2013) Caspase inhibitors: prospective therapies for stroke. Neuroscientist 19(2):129–136. https://doi.org/10.1177/1073858412447875
Bao Y, Lin C, Ren J, Liu J (2013) MicroRNA-384-5p regulates ischemia-induced cardioprotection by targeting phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit delta (PI3K p110δ). Apoptosis 18(3):260–270. https://doi.org/10.1007/s10495-013-0802-1
Béjot Y, Daubail B, Jacquin A, Durier J, Osseby GV, Rouaud O, Giroud M (2014) Trends in the incidence of ischaemic stroke in young adults between 1985 and 2011: the Dijon Stroke Registry. J Neurol Neurosurg Psychiatry 85(5):509–513. https://doi.org/10.1136/jnnp-2013-306203
Broughton BR, Reutens DC, Sobey CG (2009) Apoptotic mechanisms after cerebral ischemia. Stroke 40(5):e331-339. https://doi.org/10.1161/strokeaha.108.531632
Brouns R, De Deyn PP (2009) The complexity of neurobiological processes in acute ischemic stroke. Clin Neurol Neurosurg 111(6):483–495. https://doi.org/10.1016/j.clineuro.2009.04.001
Chen ZX, He D, Mo QW, Xie LP, Liang JR, Liu L, Fu WJ (2020) MiR-129-5p protects against myocardial ischemia-reperfusion injury via targeting HMGB1. Eur Rev Med Pharmacol Sci 24(8):4440–4450. https://doi.org/10.26355/eurrev_202004_21026
Collaborators GN (2019) Global, regional, and national burden of neurological disorders, 1990–2016: a systematic analysis for the global burden of disease study 2016. Lancet Neurol 18(5):459–480. https://doi.org/10.1016/s1474-4422(18)30499-x
Dyrskjøt L, Ostenfeld MS, Bramsen JB, Silahtaroglu AN, Lamy P, Ramanathan R, Orntoft TF (2009) Genomic profiling of microRNAs in bladder cancer: miR-129 is associated with poor outcome and promotes cell death in vitro. Cancer Res 69(11):4851–4860. https://doi.org/10.1158/0008-5472.Can-08-4043
Fan J, Xu W, Nan S, Chang M, Zhang Y (2020) MicroRNA-384-5p promotes endothelial progenitor cell proliferation and angiogenesis in cerebral ischemic stroke through the delta-likeligand 4-mediated notch signaling pathway. Cerebrovasc Dis 49(1):39–54. https://doi.org/10.1159/000503950
Fang J, Song XW, Tian J, Chen HY, Li DF, Wang JF, Lin L (2012) Overexpression of microRNA-378 attenuates ischemia-induced apoptosis by inhibiting caspase-3 expression in cardiac myocytes. Apoptosis 17(4):410–423. https://doi.org/10.1007/s10495-011-0683-0
Goh SY, Chao YX, Dheen ST, Tan EK, Tay SS (2019) Role of microRNAs in Parkinson’s disease. Int J Mol Sci. https://doi.org/10.3390/ijms20225649
Idda ML, Munk R, Abdelmohsen K, Gorospe M (2018) Noncoding RNAs in Alzheimer’s disease. Wiley Interdiscip Rev RNA. https://doi.org/10.1002/wrna.1463
Karaayvaz M, Zhai H, Ju J (2013) miR-129 promotes apoptosis and enhances chemosensitivity to 5-fluorouracil in colorectal cancer. Cell Death Dis 4:e659. https://doi.org/10.1038/cddis.2013.193
Khoshnam SE, Winlow W, Farzaneh M, Farbood Y, Moghaddam HF (2017) Pathogenic mechanisms following ischemic stroke. Neurol Sci 38(7):1167–1186. https://doi.org/10.1007/s10072-017-2938-1
Li M, Li Y, Sun L, Song JL, Lv C (2018) High mobility group box 1 promotes apoptosis of astrocytes after oxygen glucose deprivation/reoxygenation by regulating the expression of Bcl-2 and Bax. Beijing Da Xue Xue Bao Yi Xue Ban 50(5):785–791. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/30337736
Liu J, An P, Xue Y, Che D, Liu X, Zheng J, Yu B (2019) Mechanism of Snhg8/miR-384/Hoxa13/FAM3A axis regulating neuronal apoptosis in ischemic mice model. Cell Death Dis 10(6):441. https://doi.org/10.1038/s41419-019-1631-0
Ma R, Chen X, Ma Y, Bai G, Li DS (2020) MiR-129-5p alleviates myocardial injury by targeting suppressor of cytokine signaling 2 after ischemia/reperfusion. Kaohsiung J Med Sci 36(8):599–606. https://doi.org/10.1002/kjm2.12211
Mo ZT, Li WN, Zhai YR, Gong QH (2016) Icariin attenuates OGD/R-induced autophagy via Bcl-2-dependent cross talk between apoptosis and autophagy in PC12 Cells. Evid Based Complement Alternat Med 2016:4343084. https://doi.org/10.1155/2016/4343084
Nam JW, Rissland OS, Koppstein D, Abreu-Goodger C, Jan CH, Agarwal V, Bartel DP (2014) Global analyses of the effect of different cellular contexts on microRNA targeting. Mol Cell 53(6):1031–1043. https://doi.org/10.1016/j.molcel.2014.02.013
Palumbo S, Miracco C, Pirtoli L, Comincini S (2014) Emerging roles of microRNA in modulating cell-death processes in malignant glioma. J Cell Physiol 229(3):277–286. https://doi.org/10.1002/jcp.24446
Qiu J, Zhou XY, Zhou XG, Cheng R, Liu HY, Li Y (2013) Neuroprotective effects of microRNA-210 against oxygen-glucose deprivation through inhibition of apoptosis in PC12 cells. Mol Med Rep 7(6):1955–9. https://doi.org/10.3892/mmr.2013.1431
Rami A, Bechmann I, Stehle JH (2008) Exploiting endogenous anti-apoptotic proteins for novel therapeutic strategies in cerebral ischemia. Prog Neurobiol 85(3):273–296. https://doi.org/10.1016/j.pneurobio.2008.04.003
Shi H, Sun BL, Zhang J, Lu S, Zhang P, Wang H, Gao Y (2013) miR-15b suppression of Bcl-2 contributes to cerebral ischemic injury and is reversed by sevoflurane preconditioning. CNS Neurol Disord Drug. Targets 12(3):381–91. https://doi.org/10.2174/1871527311312030011
Sun Y, Xu Y, Geng L (2015) Caspase-3 inhibitor prevents the apoptosis of brain tissue in rats with acute cerebral infarction. Exp Ther Med 10(1):133–138. https://doi.org/10.3892/etm.2015.2462
Vasudeva K, Munshi A (2020) miRNA dysregulation in ischaemic stroke: focus on diagnosis, prognosis, therapeutic and protective biomarkers. Eur J Neurosci 52(6):3610–3627. https://doi.org/10.1111/ejn.14695
Wang L, Sun J, Cao H (2019) MicroRNA-384 regulates cell proliferation and apoptosis through directly targeting WISP1 in laryngeal cancer. J Cell Biochem 120(3):3018–3026. https://doi.org/10.1002/jcb.27323
Wei Q, Zhou HY, Shi XD, Cao HY, Qin L (2019) Long noncoding RNA NEAT1 promotes myocardiocyte apoptosis and suppresses proliferation through regulation of miR-129-5p. J Cardiovasc Pharmacol 74(6):535–541. https://doi.org/10.1097/fjc.0000000000000741
Xing J, Liu J, Xu Z (2020) miR-129-5p ameliorates ischemia-reperfusion injury by targeting HMGB1 in myocardium. Gen Physiol Biophys 39(5):461–470. https://doi.org/10.4149/gpb_2020021
Xue F, Cheng Y, Xu L, Tian C, Jiao H, Wang R, Gao X (2020) LncRNA NEAT1/miR-129/Bcl-2 signaling axis contributes to HDAC inhibitor tolerance in nasopharyngeal cancer. Aging (albany NY) 12(14):14174–14188. https://doi.org/10.18632/aging.103427
Yuan J, Zeng L, Sun Y, Wang N, Sun Q, Cheng Z, Wang Y (2018) SH2B1 protects against OGD/R-induced apoptosis in PC12 cells via activation of the JAK2/STAT3 signaling pathway. Mol Med Rep 18(3):2613–2620. https://doi.org/10.3892/mmr.2018.9265
Zhai F, Zhang X, Guan Y, Yang X, Li Y, Song G, Guan L (2012) Expression profiles of microRNAs after focal cerebral ischemia/reperfusion injury in rats. Neural Regen Res 7(12):917–923. https://doi.org/10.3969/j.issn.1673-5374.2012.12.007
Zhang C, Liang R, Gan X, Yang X, Chen L, Jian J (2019a) MicroRNA-384-5p/Beclin-1 As potential indicators for epigallocatechin gallate against cardiomyocytes ischemia reperfusion injury by inhibiting autophagy Via PI3K/Akt pathway. Drug Des Devel Ther 13:3607–3623. https://doi.org/10.2147/dddt.S219074
Zhang ZB, Tan YX, Zhao Q, Xiong LL, Liu J, Xu FF, Wang TH (2019b) miRNA-7a-2-3p inhibits neuronal apoptosis in oxygen-glucose deprivation (OGD) model. Front Neurosci 13:16. https://doi.org/10.3389/fnins.2019.00016
Zheng J, Liu X, Wang P, Xue Y, Ma J, Qu C, Liu Y (2016) CRNDE promotes malignant progression of glioma by attenuating miR-384/PIWIL4/STAT3 axis. Mol Ther 24(7):1199–1215. https://doi.org/10.1038/mt.2016.71
Acknowledgements
We thank all colleagues from the departments of all authors in this paper.
Funding
The project was financially supported by the National Natural Science Foundation of China, No. 81973317, No. 81374007, No. 81870977; the Natural Science Foundation of Heilongjiang Province, No. HL2019H062; the Projects of Basic Scientific Research Business Expenses in Higher Education Institutions of Heilongjiang Province,No.2018-KYYWF-MY-005; and the Students Innovative and Entrepreneurship Training Scientific Research Foundation of Heilongjiang Province, No.102292017001.
Author information
Authors and Affiliations
Contributions
H-L G, L-X G and F-G Z designed the experiments, performed the experiments and wrote the manuscript. Y G, W-Y L, J-W L, Y-J Z, Y-Z G, H-F L performed the experiments, collected the data and analyzed the data. All authors have read and approved the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no conflict of any financial AND non-financial interests.
Ethical approval
The study was approved by the Institutional Review Board of Mudanjiang Medical University.
Additional information
Communicated by Sreedharan Sajikumar.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
221_2021_6236_MOESM1_ESM.tif
Supplementary file1 Supplementary Figure 1 A The structure of bcl-2 wild-type and bcl-2 mutant vector. Potential binding site between miR-129 and bcl-2 mRNA 3'-UTR, which is between miR-129 and bcl-2 3'-UTR target region, Inter-complementary nucleotides are connected by short vertical lines. B The structure of caspase-3 wild-type and caspase-3 mutant vector. Potential binding site between miR-384-5p and caspase-3 mRNA 3'-UTR. The complementary nucleotides between miR-384-5p and caspase-3 3'-UTR target regions are connected by short vertical lines. (TIF 4831 KB)
Rights and permissions
About this article
Cite this article
Guan, HL., Guan, Y., Li, WY. et al. Regulatory role of miR-129 and miR-384-5p on apoptosis induced by oxygen and glucose deprivation in PC12 cell. Exp Brain Res 240, 97–111 (2022). https://doi.org/10.1007/s00221-021-06236-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00221-021-06236-z