The Cerebellum

, Volume 17, Issue 4, pp 477–484 | Cite as

Inhibition of Inducible Nitric Oxide Synthase Attenuates Deficits in Synaptic Plasticity and Brain Functions Following Traumatic Brain Injury

  • Bo WangEmail author
  • Shuangshuang Han
Original Paper


Traumatic brain injury (TBI), resulting from external force on the head, usually leads to long-term deficits in motor and cognitive functions. Inducible nitric oxide synthase (iNOS)-mediated excessive inflammation could exacerbate brain damage after TBI. The present study therefore investigated the potential neuroprotective effects of iNOS inhibition after TBI. Male C57BL/6J mice were subjected to controlled cortical impact injury and then treated with high selective iNOS inhibitor 1400W. Expression of iNOS mRNA was determined by quantitative RT-PCR. Western blotting was carried out to determine iNOS protein levels. Motor and cognitive functions, and long-term potentiation (LTP) in the medial prefrontal cortex (mPFC) and hippocampus were examined. Expression of iNOS was induced after TBI in a temporal manner. Treatment with 1400W after TBI improved motor and cognitive functions. TBI mice showed deficits in LTP in both the mPFC and hippocampus, and treatment with 1400W could rescue this impairment. Inhibition of iNOS attenuated deficits in synaptic plasticity and brain functions after TBI. The neuroprotective effect of iNOS inhibition on cognitive function might be via rescuing the TBI-induced LTP impairment.


Traumatic brain injury (TBI) Neuroprotection Inhibitor Inducible nitric oxide synthase (iNOS) Behaviors 


Compliance with Ethical Standards

Research Involving Human Participants and/or Animals

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Informed Consent

Not applicable.

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12311_2018_934_MOESM1_ESM.docx (143 kb)
Supplemental Figure S1 (DOCX 142 kb)


  1. 1.
    Alves JL. Blood-brain barrier and traumatic brain injury. J Neurosci Res. 2014;92:141–7. Scholar
  2. 2.
    Thal SC, Neuhaus W. The Blood–brain barrier as a target in traumatic brain injury treatment. Arch Med Res. 2014;45:698–710. Scholar
  3. 3.
    Cederberg D, Siesjo P. What has inflammation to do with traumatic brain injury? Child’s Nerv Syst. 2010;26:221–6. Scholar
  4. 4.
    Zamora R, Vodovotz Y, Billiar TR. Inducible nitric oxide synthase and inflammatory diseases. Mol Med. 2000;6:347–73.PubMedPubMedCentralCrossRefGoogle Scholar
  5. 5.
    Garry PS, Ezra M, Rowland MJ, Westbrook J, Pattinson KTS. The role of the nitric oxide pathway in brain injury and its treatment—from bench to bedside. Exp Neurol. 2015;263:235–43. Scholar
  6. 6.
    Marletta MA. Mammalian synthesis of nitrite, nitrate, nitric oxide, and N-nitrosating agents. Chem Res Toxicol. 1988;1:249–57.CrossRefPubMedGoogle Scholar
  7. 7.
    Nathan C, Calingasan N, Nezezon J, Ding A, Lucia MS, La Perle K, et al. Protection from Alzheimer’s-like disease in the mouse by genetic ablation of inducible nitric oxide synthase. J Exp Med. 2005;202:1163–9. Scholar
  8. 8.
    Gahm C, Holmin S, Wiklund PN, Brundin L, Mathiesen T. Neuroprotection by selective inhibition of inducible nitric oxide synthase after experimental brain contusion. J Neurotrauma. 2006;23:1343–54. Scholar
  9. 9.
    Schinzel R and Tegtmeier F. Chapter 8—nitric oxide synthase inhibitors in traumatic brain injury A2—Heidenreich, Kim A. New therapeutics for traumatic brain injury. San Diego, Academic Press; 2017. pp. 133–144.Google Scholar
  10. 10.
    Iadecola C, Zhang F, Xu X. Inhibition of inducible nitric oxide synthase ameliorates cerebral ischemic damage. Am J Phys. 1995;268:R286–92.Google Scholar
  11. 11.
    Cockroft KM, Meistrell M 3rd, Zimmerman GA, Risucci D, Bloom O, Cerami A, et al. Cerebroprotective effects of aminoguanidine in a rodent model of stroke. Stroke. 1996;27:1393–8.CrossRefPubMedGoogle Scholar
  12. 12.
    Iadecola C, Zhang F, Casey R, Nagayama M, Ross ME. Delayed reduction of ischemic brain injury and neurological deficits in mice lacking the inducible nitric oxide synthase gene. The Journal of neuroscience : the official journal of the Society for Neuroscience. 1997;17:9157–64.CrossRefGoogle Scholar
  13. 13.
    Stover JF, Belli A, Boret H, Bulters D, Sahuquillo J, Schmutzhard E, et al. Nitric oxide synthase inhibition with the antipterin VAS203 improves outcome in moderate and severe traumatic brain injury: a placebo-controlled randomized phase IIa trial (NOSTRA). J Neurotrauma. 2014;31:1599–606. Scholar
  14. 14.
    Xie QW, Kashiwabara Y, Nathan C. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase. J Biol Chem. 1994;269:4705–8.PubMedGoogle Scholar
  15. 15.
    Hu YC, Sun Q, Li W, Zhang DD, Ma B, Li S, et al. Biphasic activation of nuclear factor kappa B and expression of p65 and c-Rel after traumatic brain injury in rats. Inflamm Res. 2014;63:109–15. Scholar
  16. 16.
    Hwang S-Y, Hwang J-S, Kim S-Y, Han I-O. O-GlcNAcylation and p50/p105 binding of c-Rel are dynamically regulated by LPS and glucosamine in BV2 microglia cells. Br J Pharmacol. 2013;169:1551–60. Scholar
  17. 17.
    Shono Y, Tuckett AZ, Liou HC, Doubrovina E, Derenzini E, Ouk S, et al. Characterization of a c-Rel inhibitor that mediates anticancer properties in hematologic malignancies by blocking NF-kappaB-controlled oxidative stress responses. Cancer Res. 2016;76:377–89. Scholar
  18. 18.
    Garvey EP, Oplinger JA, Furfine ES, Kiff RJ, Laszlo F, Whittle BJ, et al. 1400W is a slow, tight binding, and highly selective inhibitor of inducible nitric-oxide synthase in vitro and in vivo. J Biol Chem. 1997;272:4959–63.CrossRefPubMedGoogle Scholar
  19. 19.
    Jafarian-Tehrani M, Louin G, Royo NC, Besson VC, Bohme GA, Plotkine M, et al. 1400W, a potent selective inducible NOS inhibitor, improves histopathological outcome following traumatic brain injury in rats. Nitric Oxide. 2005;12:61–9. Scholar
  20. 20.
    Laskowitz DT, Wang H, Chen T, Lubkin DT, Cantillana V, Tu TM, et al. Neuroprotective pentapeptide CN-105 is associated with reduced sterile inflammation and improved functional outcomes in a traumatic brain injury murine model. Sci Rep. 2017;7:46461. Scholar
  21. 21.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods (San Diego, Calif). 2001;25:402–8. Scholar
  22. 22.
    Grover LM, Kim E, Cooke JD, Holmes WR. LTP in hippocampal area CA1 is induced by burst stimulation over a broad frequency range centered around delta. Learning & memory (Cold Spring Harbor, NY). 2009;16:69–81. Scholar
  23. 23.
    Wang ZM, Qi YJ, Wu PY, Zhu Y, Dong YL, Cheng ZX, et al. Neuroactive steroid pregnenolone sulphate inhibits long-term potentiation via activation of alpha2-adrenoreceptors at excitatory synapses in rat medial prefrontal cortex. Int J Neuropsychopharmacol. 2008;11:611–24. Scholar
  24. 24.
    Malinow R. Transmission between pairs of hippocampal slice neurons: quantal levels, oscillations, and LTP. Science (New York, NY). 1991;252:722–4.CrossRefGoogle Scholar
  25. 25.
    Petrov T, Page AB, Owen CR, Rafols JA. Expression of the inducible nitric oxide synthase in distinct cellular types after traumatic brain injury: an in situ hybridization and immunocytochemical study. Acta Neuropathol. 2000;100:196–204. Scholar
  26. 26.
    Sinz EH, Kochanek PM, Dixon CE, Clark RS, Carcillo JA, Schiding JK, et al. Inducible nitric oxide synthase is an endogenous neuroprotectant after traumatic brain injury in rats and mice. J Clin Invest. 1999;104:647–56. Scholar
  27. 27.
    Assreuy J, Cunha FQ, Liew FY, Moncada S. Feedback inhibition of nitric oxide synthase activity by nitric oxide. Br J Pharmacol. 1993;108:833–7.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Katsumoto S, Smith SME, Martasek P, Salerno JC. Competition and binding of arginine, imidazole, and aminoguanidine to endothelial nitric oxide synthase: aminoguanidine is a poor model for substrate, intermediate, and arginine analog inhibitor binding. Nitric Oxide. 2003;8:149–54. Scholar
  29. 29.
    Vitecek J, Lojek A, Valacchi G, Kubala L. Arginine-based inhibitors of nitric oxide synthase: therapeutic potential and challenges. Mediat Inflamm. 2012;2012:318087–22. Scholar
  30. 30.
    Cobbs CS, Fenoy A, Bredt DS, Noble LJ. Expression of nitric oxide synthase in the cerebral microvasculature after traumatic brain injury in the rat. Brain Res. 1997;751:336–8.CrossRefPubMedGoogle Scholar
  31. 31.
    Huang Z, Huang PL, Ma J, Meng W, Ayata C, Fishman MC, et al. Enlarged infarcts in endothelial nitric oxide synthase knockout mice are attenuated by nitro-L-arginine. J Cerebral Blood Flow Metab. 1996;16:981–7. Scholar
  32. 32.
    Connor JR, Manning PT, Settle SL, Moore WM, Jerome GM, Webber RK, et al. Suppression of adjuvant-induced arthritis by selective inhibition of inducible nitric oxide synthase. Eur J Pharmacol. 1995;273:15–24. Scholar
  33. 33.
    Louin G, Marchand-Verrecchia C, Palmier B, Plotkine M, Jafarian-Tehrani M. Selective inhibition of inducible nitric oxide synthase reduces neurological deficit but not cerebral edema following traumatic brain injury. Neuropharmacology. 2006;50:182–90. Scholar
  34. 34.
    Shors TJ, Matzel LD. Long-term potentiation: what’s learning got to do with it? Behav Brain Sci. 1997;20:597–614. discussion -55PubMedGoogle Scholar
  35. 35.
    Togashi H, Ueno K-I, Mori K, Matsumoto M, Itoh Y, Shinohara K, et al. Nitric oxide production and long-term potentiation in the rat hippocampus following transient cerebral ischemia. In: Kitabatake A, Sakuma I, editors. Recent advances in nitric oxide research. Tokyo: Springer; 1999. p. 53–65.CrossRefGoogle Scholar
  36. 36.
    Barker GRI, Warburton EC. When is the hippocampus involved in recognition memory? J Neurosci. 2011;31:10721–31.CrossRefPubMedGoogle Scholar
  37. 37.
    Barker GRI, Bird F, Alexander V, Warburton EC. Recognition memory for objects, place, and temporal order: a disconnection analysis of the role of the medial prefrontal cortex and perirhinal cortex. J Neurosci. 2007;27:2948–57.CrossRefPubMedGoogle Scholar
  38. 38.
    Morris RG, Garrud P, Rawlins JN, O’Keefe J. Place navigation impaired in rats with hippocampal lesions. Nature. 1982;297:681–3.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Emergency DepartmentCangzhou Central HospitalCangzhouChina
  2. 2.Cangzhou Medical CollegeCangzhouChina

Personalised recommendations