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Journal of Bioenergetics and Biomembranes

, Volume 51, Issue 4, pp 291–300 | Cite as

MicroRNA-141 protects PC12 cells against hypoxia/reoxygenation-induced injury via regulating Keap1-Nrf2 signaling pathway

  • Bin Zhou
  • Hong-Yun Liu
  • Bao-Lian Zhu
  • Ai-Xia YueEmail author
Article
  • 175 Downloads

Abstract

To understand the role of microRNA-141 (miR-141) in hypoxia/reoxygenation (H/R)-induced PC12 cell injury via modulation of Keap1/Nrf2 signaling pathway. PC12 cells were divided into Control, H/R, H/R + miR-141 mimics, H/R + NC, H/R + miR-141 inhibitor, H/R + siKeap1 and H/R + miR-141 inhibitors+siKeap1 groups. The expression of miR-141 and Keap1/Nrf2 pathway was measured by qRT-PCR and western blotting, cell viability evaluated by MTT assay while cell apoptosis tested by flow cytometry. Besides, MDA (malondialdehyde), SOD (Super Oxide Dismutase) and LDH (lactate dehydrogenase) levels were determined. DCFH-DA and JC-1 staining were used to measure ROS and mitochondrial membrane potential (MMP) respectively. Compared with Controls, PC12 cells induced by H/R exhibited decreased cell viability and increased cell apoptosis rate, with elevated MDA, LDH and ROS and reduced SOD levels; and meanwhile, MMP and miR-141 expression were declined, whereas cytoplasmic Nrf2 levels were enhanced with the downregulated nuclear Nrf2 level (all P < 0.05). However, these cells treated with miR-141 mimics and siKeap1 showed obvious improvement in H/R-induced cell injury, while miR-141 inhibitors presented significantly aggravated cell injury (both P < 0.05). Besides, siKeap1 can reverse the effect of miRNA-141 inhibitors on aggravating H/R-induced PC12 cell injury. miR-141-mediated Keap1/Nrf2 signaling pathway to promote cell viability, inhibit cell apoptosis and reduce oxidative stress of PC12 cells, thereby alleviating H/R-induced cell injury.

Keywords

miR-141 Keap1/Nrf2 Hypoxia/reoxygenation PC12 cell 

Notes

Acknowledgements

We are grateful to all the reviewers for their comments for this work.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest regarding the publication of this study.

References

  1. Calabrese V, Cornelius C, Mancuso C, Lentile R, Stella AM, Butterfield DA (2010) Redox homeostasis and cellular stress response in aging and neurodegeneration. Methods Mol Biol 610:285–308.  https://doi.org/10.1007/978-1-60327-029-8_17 CrossRefPubMedGoogle Scholar
  2. Candelario-Jalil E (2009) Injury and repair mechanisms in ischemic stroke: considerations for the development of novel neurotherapeutics. Curr Opin Investig Drugs 10(7):644–654PubMedGoogle Scholar
  3. Cao X, Xiao H, Zhang Y, Zou L, Chu Y, Chu X (2010) 1, 5-Dicaffeoylquinic acid-mediated glutathione synthesis through activation of Nrf2 protects against OGD/reperfusion-induced oxidative stress in astrocytes. Brain Res 1347:142–148.  https://doi.org/10.1016/j.brainres.2010.05.072 CrossRefPubMedGoogle Scholar
  4. Castellano L, Stebbing J (2013) Deep sequencing of small RNAs identifies canonical and non-canonical miRNA and endogenous siRNAs in mammalian somatic tissues. Nucleic Acids Res 41(5):3339–3351.  https://doi.org/10.1093/nar/gks1474 CrossRefPubMedPubMedCentralGoogle Scholar
  5. Chan YC, Khanna S, Roy S, Sen CK (2011) miR-200b targets Ets-1 and is down-regulated by hypoxia to induce angiogenic response of endothelial cells. J Biol Chem 286(3):2047–2056.  https://doi.org/10.1074/jbc.M110.158790 CrossRefPubMedGoogle Scholar
  6. Cheng LB, Li KR, Yi N, Li XM, Wang F, Xue B et al (2017) miRNA-141 attenuates UV-induced oxidative stress via activating Keap1-Nrf2 signaling in human retinal pigment epithelium cells and retinal ganglion cells. Oncotarget 8(8):13186–13194.  https://doi.org/10.18632/oncotarget.14489 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Ciancarelli I, Di Massimo C, De Amicis D, Pistarini C, Tozzi Ciancarelli MG (2015) Uric acid and cu/Zn superoxide dismutase: potential strategies and biomarkers in functional recovery of post-acute ischemic stroke patients after intensive neurorehabilitation. Curr Neurovasc Res 12(2):120–127CrossRefPubMedGoogle Scholar
  8. Cui C, Cui N, Wang P, Song S, Liang H, Ji A (2015) Sulfated polysaccharide isolated from the sea cucumber Stichopus japonicus against PC12 hypoxia/Reoxygenation injury by inhibition of the MAPK signaling pathway. Cell Mol Neurobiol 35(8):1081–1092.  https://doi.org/10.1007/s10571-015-0202-x CrossRefPubMedGoogle Scholar
  9. Eltzschig HK, Eckle T (2011) Ischemia and reperfusion--from mechanism to translation. Nat Med 17(11):1391–1401.  https://doi.org/10.1038/nm.2507 CrossRefPubMedGoogle Scholar
  10. Feigin VL, Lawes CM, Bennett DA, Barker-Collo SL, Parag V (2009) Worldwide stroke incidence and early case fatality reported in 56 population-based studies: a systematic review. Lancet Neurol 8(4):355–369.  https://doi.org/10.1016/S1474-4422(09)70025-0 CrossRefPubMedGoogle Scholar
  11. Grau CM, Greene LA (2012) Use of PC12 cells and rat superior cervical ganglion sympathetic neurons as models for neuroprotective assays relevant to Parkinson's disease. Methods Mol Biol 846:201–211.  https://doi.org/10.1007/978-1-61779-536-7_18 CrossRefPubMedPubMedCentralGoogle Scholar
  12. Kang MI, Kobayashi A, Wakabayashi N, Kim SG, Yamamoto M (2004) Scaffolding of Keap1 to the actin cytoskeleton controls the function of Nrf2 as key regulator of cytoprotective phase 2 genes. Proc Natl Acad Sci U S A 101(7):2046–2051.  https://doi.org/10.1073/pnas.0308347100 CrossRefPubMedPubMedCentralGoogle Scholar
  13. Kensler TW, Wakabayashi N, Biswal S (2007) Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. Annu Rev Pharmacol Toxicol 47:89–116.  https://doi.org/10.1146/annurev.pharmtox.46.120604.141046 CrossRefPubMedGoogle Scholar
  14. Kulisz A, Chen N, Chandel NS, Shao Z, Schumacker PT (2002) Mitochondrial ROS initiate phosphorylation of p38 MAP kinase during hypoxia in cardiomyocytes. Am J Phys Lung Cell Mol Phys 282(6):L1324–L1329.  https://doi.org/10.1152/ajplung.00326.2001 CrossRefGoogle Scholar
  15. Lau A, Villeneuve NF, Sun Z, Wong PK, Zhang DD (2008) Dual roles of Nrf2 in cancer. Pharmacol Res 58(5–6):262–270.  https://doi.org/10.1016/j.phrs.2008.09.003 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Li H, Tang J, Lei H, Cai P, Zhu H, Li B, Xu X, Xia Y, Tang W (2014) Decreased MiR-200a/141 suppress cell migration and proliferation by targeting PTEN in Hirschsprung's disease. Cell Physiol Biochem 34(2):543–553.  https://doi.org/10.1159/000363021 CrossRefPubMedGoogle Scholar
  17. Li Y, Zhu X, Liu X, Du A, Yu B (2017) MiR-200a mediates protection of thymosin beta 4 in cardiac microvascular endothelial cells as a novel mechanism under hypoxia-reoxygenation injury. J Cell Biochem 120:12069.  https://doi.org/10.1002/jcb.26381 CrossRefGoogle Scholar
  18. Liu RR, Li J, Gong JY, Kuang F, Liu JY, Zhang YS, Ma QL, Song C, Truax AD, Gao F, Yang K, Jin BQ, Chen LH (2015) MicroRNA-141 regulates the expression level of ICAM-1 on endothelium to decrease myocardial ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol 309(8):H1303–H1313.  https://doi.org/10.1152/ajpheart.00290.2015 CrossRefPubMedGoogle Scholar
  19. Mironov SL (2009) Complexity of mitochondrial dynamics in neurons and its control by ADP produced during synaptic activity. Int J Biochem Cell Biol 41(10):2005–2014.  https://doi.org/10.1016/j.biocel.2009.04.009 CrossRefPubMedGoogle Scholar
  20. Naziroglu M, Kutluhan S, Uguz AC, Celik O, Bal R, Butterworth PJ (2009) Topiramate and vitamin e modulate the electroencephalographic records, brain microsomal and blood antioxidant redox system in pentylentetrazol-induced seizure of rats. J Membr Biol 229(3):131–140.  https://doi.org/10.1007/s00232-009-9177-1 CrossRefPubMedGoogle Scholar
  21. Pieraccioli M, Imbastari F, Antonov A, Melino G, Raschella G (2013) Activation of miR200 by c-Myb depends on ZEB1 expression and miR200 promoter methylation. Cell Cycle 12(14):2309–2320.  https://doi.org/10.4161/cc.25405 CrossRefPubMedPubMedCentralGoogle Scholar
  22. Rocha O, Ansari K, Doohan FM (2005) Effects of trichothecene mycotoxins on eukaryotic cells: a review. Food Addit Contam 22(4):369–378.  https://doi.org/10.1080/02652030500058403 CrossRefPubMedGoogle Scholar
  23. Shi L, Wu L, Chen Z, Yang J, Chen X, Yu F, Zheng F, Lin X (2015) MiR-141 activates Nrf2-dependent antioxidant pathway via Down-regulating the expression of Keap1 conferring the resistance of hepatocellular carcinoma cells to 5-fluorouracil. Cell Physiol Biochem 35(6):2333–2348.  https://doi.org/10.1159/000374036 CrossRefPubMedGoogle Scholar
  24. Stewart JD, Hengstler JG, Bolt HM (2011) Control of oxidative stress by the Keap1-Nrf2 pathway. Arch Toxicol 85(4):239.  https://doi.org/10.1007/s00204-011-0694-1 CrossRefPubMedGoogle Scholar
  25. Sun X, Zuo H, Liu C, Yang Y (2016) Overexpression of miR-200a protects cardiomyocytes against hypoxia-induced apoptosis by modulating the kelch-like ECH-associated protein 1-nuclear factor erythroid 2-related factor 2 signaling axis. Int J Mol Med 38(4):1303–1311.  https://doi.org/10.3892/ijmm.2016.2719 CrossRefPubMedGoogle Scholar
  26. Tang H, Deng M, Tang Y, Xie X, Guo J, Kong Y, Ye F, Su Q, Xie X (2013) miR-200b and miR-200c as prognostic factors and mediators of gastric cancer cell progression. Clin Cancer Res 19(20):5602–5612.  https://doi.org/10.1158/1078-0432.CCR-13-1326 CrossRefPubMedGoogle Scholar
  27. Tegos TJ, Kalodiki E, Daskalopoulou SS, Nicolaides AN (2000) Stroke: epidemiology, clinical picture, and risk factors--part I of III. Angiology 51(10):793–808.  https://doi.org/10.1177/000331970005101001 CrossRefPubMedGoogle Scholar
  28. Uesugi S, Muroi M, Kondoh Y, Shiono Y, Osada H, Kimura KI (2017) Allantopyrone a activates Keap1-Nrf2 pathway and protects PC12 cells from oxidative stress-induced cell death. J Antibiot (Tokyo) 70(4):429–434.  https://doi.org/10.1038/ja.2016.99 CrossRefGoogle Scholar
  29. Um HC, Jang JH, Kim DH, Lee C, Surh YJ (2011) Nitric oxide activates Nrf2 through S-nitrosylation of Keap1 in PC12 cells. Nitric Oxide 25(2):161–168.  https://doi.org/10.1016/j.niox.2011.06.001 CrossRefPubMedGoogle Scholar
  30. van Jaarsveld MT, Helleman J, Boersma AW, van Kuijk PF, van Ijcken WF, Despierre E et al (2013) miR-141 regulates KEAP1 and modulates cisplatin sensitivity in ovarian cancer cells. Oncogene 32(36):4284–4293.  https://doi.org/10.1038/onc.2012.433 CrossRefPubMedGoogle Scholar
  31. Wang CH, Wu SB, Wu YT, Wei YH (2013) Oxidative stress response elicited by mitochondrial dysfunction: implication in the pathophysiology of aging. Exp Biol Med (Maywood) 238(5):450–460.  https://doi.org/10.1177/1535370213493069 CrossRefGoogle Scholar
  32. Wang PF, Fang H, Chen J, Lin S, Liu Y, Xiong XY, Wang YC, Xiong RP, lv FL, Wang J, Yang QW (2014) Polyinosinic-polycytidylic acid has therapeutic effects against cerebral ischemia/reperfusion injury through the downregulation of TLR4 signaling via TLR3. J Immunol 192(10):4783–4794.  https://doi.org/10.4049/jimmunol.1303108 CrossRefPubMedPubMedCentralGoogle Scholar
  33. Warach S, Latour LL (2004) Evidence of reperfusion injury, exacerbated by thrombolytic therapy, in human focal brain ischemia using a novel imaging marker of early blood-brain barrier disruption. Stroke 35(11 Suppl 1):2659–2661.  https://doi.org/10.1161/01.STR.0000144051.32131.09 CrossRefPubMedGoogle Scholar
  34. Wei YH, Lu CY, Lee HC, Pang CY, Ma YS (1998) Oxidative damage and mutation to mitochondrial DNA and age-dependent decline of mitochondrial respiratory function. Ann N Y Acad Sci 854:155–170CrossRefPubMedGoogle Scholar
  35. Yin F, Boveris A, Cadenas E (2014) Mitochondrial energy metabolism and redox signaling in brain aging and neurodegeneration. Antioxid Redox Signal 20(2):353–371.  https://doi.org/10.1089/ars.2012.4774 CrossRefPubMedPubMedCentralGoogle Scholar
  36. Zhang DD (2006) Mechanistic studies of the Nrf2-Keap1 signaling pathway. Drug Metab Rev 38(4):769–789.  https://doi.org/10.1080/03602530600971974 CrossRefPubMedGoogle Scholar
  37. Zhou Y, Wang HD, Zhu L, Cong ZX, Li N, Ji XJ et al (2013) Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line. Oncol Rep 29(1):394–400.  https://doi.org/10.3892/or.2012.2115 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Bin Zhou
    • 1
  • Hong-Yun Liu
    • 2
  • Bao-Lian Zhu
    • 3
  • Ai-Xia Yue
    • 1
    Email author
  1. 1.Department of Rehabilitation MedicineLinyi Central HospitalLinyiChina
  2. 2.Department of GynaecologyLinyi Central HospitalLinyiChina
  3. 3.Department of InfectionLinyi Central HospitalLinyiChina

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