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Expression and regulation of miR-449a and AREG in cerebral ischemic injury

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Abstract

Rodent focal ischemia models are widely used to mimic and examine human strokes. To the best of our knowledge, no investigation has systematically examined the expression changes of microRNA (miR)-449a and Amphiregulin (AREG) as well as their biological relationship during middle cerebral artery occlusion (MCAO) and oxygen and glucose deprivation/reperfusion (OGD/R). The present study examined the histological and behavioral outcomes of MCAO and the function of miR-449a and AREG in cerebral ischemic injury. Rats were subjected to 2 h MCAO, which was followed by reperfusion. miR-449a and AREG were examined in the injury tissues of MCAO rats and the OGD/R cell line by reverse transcription-quantitative polymerase chain reaction. Protein expressions of AREG in the injury tissues of MCAO rats was measured using an immunohistochemistry and the protein expression levels of AREG, epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (Akt) and the phosphorylation level of Akt (p-Akt) were analyzed by western blotting. Cell apoptosis was examined following the knock down and subsequent overexpression of AREG in a human OGD/R neuronal cell line by small interfering RNAs (siRNAs) and plasmid transfection. Luciferase reporter assays were used to validate the target of miR-449a. The expression changes and regulatory mechanisms of miR-449a and AREG in an ischemia/reperfusion (I/R) injury model were examined in vivo and in vitro. The neurological deficit score, brain edema volume, cerebral infarct area, and the number of apoptosis cells in ischemic rats were all markedly elevated, than that in the control rats. The expression of miR-449a was decreased and AREG was increased in the MCAO rats and human OGD/R neuronal cell line. miR-449a inhibition or AREG overexpression in OGD/R cells resulted in a significant decrease in apoptotic cells, and AREG was revealed to be one of the direct targets of miR-449a. Molecular recovery was observed following transfection with miR-449a mimics and AREG knockdown in an OGD/R model in vitro. The present study demonstrated that miR-449a was downregulated while AREG was upregulated in cerebral ischemic injury, and the recovery of neurological function can be obtained following the overexpression of miR-449a and the knockdown of AREG in an I/R injury model. miR-449a functions in ischemic stroke via directly targeting AREG. These findings suggest a novel mechanism involving in cerebral I/R injury model and may aid investigators in gaining a deeper understanding of strokes in a clinical setting.

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References

  • Ambros V (2004) The functions of animal microRNAs. Nature 431:350

    Article  CAS  PubMed  Google Scholar 

  • Barca-Mayo O, De Pietri Tonelli D (2014) Convergent microRNA actions coordinate neocortical development. Cell Mol Life Sci 71(16):2975–2995

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Berasain C, Avila MA (2014) Amphiregulin. Semin Cell Dev Biol 28:31–41

    Article  CAS  PubMed  Google Scholar 

  • Besancon E, Guo S, Lok J, Tymianski M, Lo EH (2008) Beyond NMDA and AMPA glutamate receptors: emerging mechanisms for ionic imbalance and cell death in stroke. Trends Pharmacol Sci 29(5):268–275

    Article  CAS  PubMed  Google Scholar 

  • Broughton BR, Reutens DC, Sobey CG (2009) Apoptotic mechanisms after cerebral ischemia. Stroke 40(5):1524–4628 (Electronic)

    Article  Google Scholar 

  • Chen X, Wu H, Chen H, Wang Q, Xie XJ (2018) Shen JA-Ohoo. Astragaloside VI Promotes Neural Stem Cell Proliferation and Enhances Neurological Function Recovery in Transient Cerebral Ischemic Injury via Activating EGFR/MAPK Signaling Cascades. https://doi.org/10.1007/s12035-018-1294-3

  • Dirnagl U, Iadecola C, Moskowitz MA (1999) Pathobiology of ischaemic stroke: an integrated view. Trends Neurosci 22(9):391–397

    Article  CAS  PubMed  Google Scholar 

  • Eckstein N, Servan K, Girard L, Cai D, von Jonquieres G, Jaehde U et al (2008) Epidermal growth factor receptor pathway analysis identifies Amphiregulin as a key factor for cisplatin resistance of human breast Cancer cells. J Biol Chem 283(2):739–750

    Article  CAS  PubMed  Google Scholar 

  • El-Marasy, SAA-Ohoo, Abdel-Rahman RF, Abd-Elsalam RM. (2018) Neuroprotective effect of vildagliptin against cerebral ischemia in rats. https://doi.org/10.1007/s00210-018-1537-x

  • Falk A, Frisen J (2002) Amphiregulin is a mitogen for adult neural stem cells. J Neurosci Res 69(6):757–762

    Article  CAS  PubMed  Google Scholar 

  • Fan J, Chen M, Wang X, Tian Z, Wang J, Fan D et al (2018) Targeting Smox is neuroprotective and ameliorates brain inflammation in cerebral ischemia/reperfusion rats. Toxicol Sci. https://doi.org/10.1093/toxsci/kfy300

  • Green AR, Shuaib A (2006) Therapeutic strategies for the treatment of stroke. Drug Discov Today 11(15):681–693

    Article  CAS  PubMed  Google Scholar 

  • Group* TNIoNDaSr-PSS (1995) Tissue plasminogen activator for acute ischemic stroke. N Engl J Med 333(24): 1581–1588

  • Huang Y, Hu ZA-Ohoo (2018) UBIAD1 protects against oxygen-glucose deprivation/reperfusion-induced multiple subcellular organelles injury through PI3K/AKT pathway in N2A cells. J Cell Physiol 233(9):7480–7496. https://doi.org/10.1002/jcp.26602

  • Kisoh K, Hayashi H, Arai M, Orita M, Yuan B, Takagi NA-Ohoo (2018) Possible Involvement of PI3-K/Akt-Dependent GSK-3beta Signaling in Proliferation of Neural Progenitor Cells After Hypoxic Exposure. https://doi.org/10.1007/s12035-018-1216-4

  • Lewis BP, Burge CB, Bartel DP (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are MicroRNA targets. Cell 120(1):15–20

    Article  CAS  PubMed  Google Scholar 

  • Longa EZ, Weinstein PR, Carlson S, Cummins R (1989) Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20(1):0039–2499

    Article  Google Scholar 

  • Lopez-Morales MA, Castello-Ruiz M, Burguete MC, Jover-Mengual T, Aliena-Valero A, Centeno JM, et al. (2018) Molecular mechanisms underlying the neuroprotective role of atrial natriuretic peptide in experimental acute ischemic stroke

  • Overgaard K, Sereghy T, Boysen G, Pedersen H, Høyer S, Diemer NH (1992) A rat model of reproducible cerebral infarction using thrombotic blood clot emboli. J Cereb Blood Flow Metab 12(3):484–490

    Article  CAS  PubMed  Google Scholar 

  • Qi D, Ouyang C, Wang Y, Zhang S, Ma X, Song Y, et al. (2014) HO-1 attenuates hippocampal neurons injury via the activation of BDNF-TrkB-PI3K/Akt signaling pathway in stroke. Brain Res 19(1577):69–76. https://doi.org/10.1016/j.brainres

  • Qian J, Wang L, Li Q, Sha D, Wang J, Zhang J et al (2019) Ultrasound-targeted microbubble enhances migration and therapeutic efficacy of marrow mesenchymal stem cell on rat middle cerebral artery occlusion stroke model. J Cell Biochem 120(3):3315–3322

    Article  CAS  Google Scholar 

  • Wu J, Bao J, Kim M, Yuan S, Tang C, Zheng H et al (2014) Two miRNA clusters, miR-34b/c and miR-449, are essential for normal brain development, motile ciliogenesis, and spermatogenesis. Proc Natl Acad Sci U S A 111(28):E2851–E2857

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yan H, Zhang X, Hu W, Ma J, Hou W, Zhang X, et al. (2014) Histamine H3 receptors aggravate cerebral ischaemic injury by histamine-independent mechanisms. Nat Commun. https://doi.org/10.1038/ncomms4334

  • Yang J, Xiong LL, Wang YC, He X, Jiang L, Fu SJ et al (2018) Oligodendrocyte precursor cell transplantation promotes functional recovery following contusive spinal cord injury in rats and is associated with altered microRNA expression. Mol Med Rep 17(1):771–782

    CAS  PubMed  Google Scholar 

  • Yao Y, Ma J, Xue Y, Wang P, Li Z, Li Z et al (2015) MiR-449a exerts tumor-suppressive functions in human glioblastoma by targeting Myc-associated zinc-finger protein. Mol Oncol 9(3):640–656

    Article  CAS  PubMed  Google Scholar 

  • Yu H, Lu Y, Li Z, Wang Q (2014) microRNA-133: expression, function and therapeutic potential in muscle diseases and cancer. Curr Drug Targets 15(9):817–828

    Article  CAS  PubMed  Google Scholar 

  • Zhang RL, Chopp M, Zhang ZG, Jiang Q, Ewing JR (1997) A rat model of focal embolic cerebral ischemia. Brain Res 766(1):83–92

    Article  CAS  PubMed  Google Scholar 

  • Zhang G, Zhang JH, Feng J, Li Q, Wu X, Qin X (2011) Electrical stimulation of olfactory bulb downregulates RGMa expression after ischemia/reperfusion injury in rats. Brain Res Bull 86(3–4):254–261

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Du Q, Yang Y, Wang J, Liu Y, Zhao Z, et al. (2014) Salidroside alleviates Ischemic Brain Injury in Mice with Ischemic Stroke through regulating BDNK mediated PI3K/Akt Pathway. https://doi.org/10.1016/j.bcp.2018.08.015

  • Zhou J, Du T, Li B, Rong Y, Verkhratsky A, Peng L. (2015) Crosstalk Between MAPK/ERK and PI3K/AKT Signal Pathways During Brain Ischemia/Reperfusion. https://doi.org/10.1177/1759091415602463

  • Zhu Q, Enkhjargal B, Huang L, Zhang T, Sun C, Xie Z, et al. (2018) Aggf1 attenuates neuroinflammation and BBB disruption via PI3K/Akt/NF-kappaB pathway after subarachnoid hemorrhage in rats. J Neuroinflammation 15(1):178. https://doi.org/10.1186/s12974-018-1211-8

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Authors and Affiliations

  1. Department of Neurology, Affiliated Hospital of ChiFeng University, ChiFeng, 024000, China

    Yanhui Yu, Zhengmin Han & Weili Zhao

  2. Department of Preventive Medicine, ChiFeng University, ChiFeng, 024000, China

    Xiuhui Zhang

  3. Department of Ophthalmology, Affiliated Hospital of ChiFeng University, ChiFeng, 024000, China

    Limin Zhang

  4. Department of Ophthalmology, Affiliated Hospital of ChiFeng University, No.4, Third section, East Garden Road, Hongshan District, Chifeng, 024005, Inner Mongolia, China

    Limin Zhang

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  1. Yanhui Yu
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Correspondence to Limin Zhang.

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Yu, Y., Zhang, X., Han, Z. et al. Expression and regulation of miR-449a and AREG in cerebral ischemic injury. Metab Brain Dis 34, 821–832 (2019). https://doi.org/10.1007/s11011-019-0393-9

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