Advertisement

Cellular and Molecular Neurobiology

, Volume 37, Issue 4, pp 587–594 | Cite as

Down-Regulation of IRF6 Protects Cortical Neurons Against Traumatic Neuronal Injury Through Activating Akt-eNOS Pathway

Original Research

Abstract

Interferon regulatory factor 6 (IRF6) is a novel and unique member of the IRF family of transcription factors, and the regulation and function of IRF6 remain unknown. Recently, IRF6 was shown to be upregulated after TBI and could promote neuronal apoptosis under oxidative stress conditions. This study aimed to investigate the role of IRF6 in traumatic neuronal injury (TNI) in primary cultured mouse cortical neurons. We found that the expression of IRF6 was significantly increased within 48 after TNI, and peaked at 24 h. Knockdown of IRF6 using specific targeted small interfering RNA (siRNA) attenuated TNI-induced loss of neuronal viability and release of lactate dehydrogenase. The results of TUNEL staining showed that IRF6 knockdown markedly reduced neuronal apoptosis, which was accompanied by decreased activity of caspase-3. Furthermore, downregulation of IRF6 inhibited lipid peroxidation, promoted the activity of endogenous antioxidative enzymes, and differently regulated the expression of inflammatory cytokines after TNI. In addition, IRF6 knockdown significantly increased phosphorylation of Akt and endothelial nitric oxide synthase (eNOS), whereas blocking Akt-eNOS pathway via selective antagonists partly prevented the protective effects of IRF6 knockdown. These data show that downregulation of IRF6 affords protection against TNI through Akt-eNOS pathway-mediated antioxidative and anti-inflammatory activity.

Keywords

IRF6 TBI Oxidative stress Akt eNOS 

References

  1. Alexander M, Forster C, Sugimoto K, Clark HB, Vogel S, Ross ME, Iadecola C (2003) Interferon regulatory factor-1 immunoreactivity in neurons and inflammatory cells following ischemic stroke in rodents and humans. Acta Neuropathol 105(5):420–424. doi: 10.1007/s00401-002-0658-x PubMedGoogle Scholar
  2. Chen W, Royer WE Jr (2010) Structural insights into interferon regulatory factor activation. Cell Signal 22(6):883–887. doi: 10.1016/j.cellsig.2009.12.005 CrossRefPubMedGoogle Scholar
  3. Chen T, Liu W, Chao X, Zhang L, Qu Y, Huo J, Fei Z (2011) Salvianolic acid B attenuates brain damage and inflammation after traumatic brain injury in mice. Brain Res Bull 84(2):163–168. doi: 10.1016/j.brainresbull.2010.11.015 CrossRefPubMedGoogle Scholar
  4. Chen T, Fei F, Jiang XF, Zhang L, Qu Y, Huo K, Fei Z (2012) Down-regulation of Homer1b/c attenuates glutamate-mediated excitotoxicity through endoplasmic reticulum and mitochondria pathways in rat cortical neurons. Free Radic Biol Med 52(1):208–217. doi: 10.1016/j.freeradbiomed.2011.10.451 CrossRefPubMedGoogle Scholar
  5. Chen T, Dai SH, Jiang ZQ, Luo P, Jiang XF, Fei Z, Gui SB, Qi YL (2016) The AMPAR Antagonist Perampanel Attenuates Traumatic Brain Injury Through Anti-Oxidative and Anti-Inflammatory Activity. Cell Mol Neurobiol. doi: 10.1007/s10571-016-0341-8 Google Scholar
  6. Chua KS, Ng YS, Yap SG, Bok CW (2007) A brief review of traumatic brain injury rehabilitation. Ann Acad Med 36(1):31–42Google Scholar
  7. Dai SH, Chen T, Wang YH, Zhu J, Luo P, Rao W, Yang YF, Fei Z, Jiang XF (2014) Sirt3 protects cortical neurons against oxidative stress via regulating mitochondrial Ca2 + and mitochondrial biogenesis. Int J Mol Sci 15(8):14591–14609. doi: 10.3390/ijms150814591 CrossRefPubMedPubMedCentralGoogle Scholar
  8. Dimmeler S, Fleming I, Fisslthaler B, Hermann C, Busse R, Zeiher AM (1999) Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature 399(6736):601–605. doi: 10.1038/21224 CrossRefPubMedGoogle Scholar
  9. Faden AI, O’Leary DM, Fan L, Bao W, Mullins PG, Movsesyan VA (2001) Selective blockade of the mGluR1 receptor reduces traumatic neuronal injury in vitro and improves outcome after brain trauma. Exp Neurol 167(2):435–444. doi: 10.1006/exnr.2000.7577 CrossRefPubMedGoogle Scholar
  10. Ferretti E, Li B, Zewdu R, Wells V, Hebert JM, Karner C, Anderson MJ, Williams T, Dixon J, Dixon MJ, Depew MJ, Selleri L (2011) A conserved Pbx-Wnt-p63-Irf6 regulatory module controls face morphogenesis by promoting epithelial apoptosis. Dev Cell 21(4):627–641. doi: 10.1016/j.devcel.2011.08.005 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Kondo S, Schutte BC, Richardson RJ, Bjork BC, Knight AS, Watanabe Y, Howard E, de Lima RL, Daack-Hirsch S, Sander A, McDonald-McGinn DM, Zackai EH, Lammer EJ, Aylsworth AS, Ardinger HH, Lidral AC, Pober BR, Moreno L, Arcos-Burgos M, Valencia C, Houdayer C, Bahuau M, Moretti-Ferreira D, Richieri-Costa A, Dixon MJ, Murray JC (2002) Mutations in IRF6 cause Van der Woude and popliteal pterygium syndromes. Nat Genet 32(2):285–289. doi: 10.1038/ng985 CrossRefPubMedPubMedCentralGoogle Scholar
  12. Lin Y, Xu D, Li X, Liu C, Liu X, Huang S, Huang Y (2015) Upregulation of interferon regulatory factor 6 promotes neuronal apoptosis after traumatic brain injury in adult rats. Cell Mol Neurobiol. doi: 10.1007/s10571-015-0217-3 Google Scholar
  13. Liu H, Liu X, Wei X, Chen L, Xiang Y, Yi F, Zhang X (2012) Losartan, an angiotensin II type 1 receptor blocker, ameliorates cerebral ischemia-reperfusion injury via PI3 K/Akt-mediated eNOS phosphorylation. Brain Res Bull 89(1–2):65–70. doi: 10.1016/j.brainresbull.2012.06.010 PubMedGoogle Scholar
  14. Maas AI, Stocchetti N, Bullock R (2008) Moderate and severe traumatic brain injury in adults. Lancet Neurol 7(8):728–741. doi: 10.1016/S1474-4422(08)70164-9 CrossRefPubMedGoogle Scholar
  15. Ooboshi H, Ibayashi S, Shichita T, Kumai Y, Takada J, Ago T, Arakawa S, Sugimori H, Kamouchi M, Kitazono T, Iida M (2005) Postischemic gene transfer of interleukin-10 protects against both focal and global brain ischemia. Circulation 111(7):913–919CrossRefPubMedGoogle Scholar
  16. Pang L, Ye W, Che XM, Roessler BJ, Betz AL, Yang GY (2001) Reduction of inflammatory response in the mouse brain with adenoviral-mediated transforming growth factor-ss1 expression. Stroke 32(2):544–552CrossRefPubMedGoogle Scholar
  17. Ray SK, Dixon CE, Banik NL (2002) Molecular mechanisms in the pathogenesis of traumatic brain injury. Histol Histopathol 17(4):1137–1152PubMedGoogle Scholar
  18. Redell JB, Moore AN, Grill RJ, Johnson D, Zhao J, Liu Y, Dash PK (2013) Analysis of functional pathways altered after mild traumatic brain injury. J Neurotrauma 30(9):752–764. doi: 10.1089/neu.2012.2437 CrossRefPubMedPubMedCentralGoogle Scholar
  19. Rochfort KD, Cummins PM (2015) The blood-brain barrier endothelium: a target for pro-inflammatory cytokines. Biochem Soc Trans 43(4):702–706. doi: 10.1042/BST20140319 CrossRefPubMedGoogle Scholar
  20. Takaoka A, Tamura T, Taniguchi T (2008) Interferon regulatory factor family of transcription factors and regulation of oncogenesis. Cancer Sci 99(3):467–478. doi: 10.1111/j.1349-7006.2007.00720.x CrossRefPubMedGoogle Scholar
  21. Vecil GG, Larsen PH, Corley SM, Herx LM, Besson A, Goodyer CG, Yong VW (2000) Interleukin-1 is a key regulator of matrix metalloproteinase-9 expression in human neurons in culture and following mouse brain trauma in vivo. J Neurosci Res 61(2):212–224CrossRefPubMedGoogle Scholar
  22. Xiang M, Wang L, Guo S, Lu YY, Lei H, Jiang DS, Zhang Y, Liu Y, Zhou Y, Zhang XD, Li H (2014) Interferon regulatory factor 8 protects against cerebral ischaemic-reperfusion injury. J Neurochem 129(6):988–1001. doi: 10.1111/jnc.12682 CrossRefPubMedGoogle Scholar
  23. Xu J, Wang H, Lu X, Ding K, Zhang L, He J, Wei W, Wu Y (2014) Posttraumatic administration of luteolin protects mice from traumatic brain injury: implication of autophagy and inflammation. Brain Res 1582:237–246. doi: 10.1016/j.brainres.2014.07.042 CrossRefPubMedGoogle Scholar
  24. Yang C, Talukder MA, Varadharaj S, Velayutham M, Zweier JL (2013) Early ischaemic preconditioning requires Akt- and PKA-mediated activation of eNOS via serine1176 phosphorylation. Cardiovasc Res 97(1):33–43. doi: 10.1093/cvr/cvs287 CrossRefPubMedGoogle Scholar
  25. Zhao GN, Jiang DS, Li H (2015) Interferon regulatory factors: at the crossroads of immunity, metabolism, and disease. Biochim Biophys Acta 2:365–378. doi: 10.1016/j.bbadis.2014.04.030 CrossRefGoogle Scholar
  26. Ziebell JM, Morganti-Kossmann MC (2010) Involvement of pro- and anti-inflammatory cytokines and chemokines in the pathophysiology of traumatic brain injury. Neurotherapeutics 7(1):22–30. doi: 10.1016/j.nurt.2009.10.016 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Neurosurgery, Shaanxi Provincial People’s Hospital, Affiliated Hospital of Xi’an Medical UniversityThe Third Affiliated Hospital of Xi’an Jiaotong UniversityXi’anChina

Personalised recommendations