Acta Neurochirurgica

, Volume 157, Issue 4, pp 649–659 | Cite as

COX-2 regulation and TUNEL-positive cell death differ between genders in the secondary inflammatory response following experimental penetrating focal brain injury in rats

  • Mattias Günther
  • Stefan Plantman
  • Johan Davidsson
  • Maria Angéria
  • Tiit Mathiesen
  • Mårten Risling
Experimental research - Brain Injury



Traumatic brain injury is followed by secondary neuronal degeneration, largely dependent on an inflammatory response. This response is probably gender specific, since females are better protected than males in experimental models. The reasons are not fully known. We examined aspects of the inflammatory response following experimental TBI in male and female rats to explore possible gender differences at 24 h and 72 h after trauma, times of peak histological inflammation and neuronal degeneration.


A penetrating brain injury model was used to produce penetrating focal TBI in 20 Sprague-Dawley rats, 5 males and 5 females for each time point. After 24 and 72 h the brains were removed and subjected to in situ hybridization and immunohistochemical analyses for COX-2, iNOS, osteopontin, glial fibrillary acidic protein, 3-nitrotyrosine, TUNEL and Fluoro-Jade.


COX-2 mRNA and protein levels were increased in the perilesional area compared to the uninjured contralateral side and significantly higher in males at 24 h and 72 h (p < 0.05). iNOS mRNA was significantly increased in females at 24 h (p < 0.05) although protein was not. TUNEL was increased in male rats after 24 h (p < 0.05). Glial fibrillary acidic protein, osteopontin, 3-nitrotyrosine and Fluoro-Jade stained degenerating neurons were increased in the perilesional area, showing no difference between genders.


COX-2 regulation differed between genders after TBI. The increased COX-2 expression in male rats correlated with increased apoptotic cell death detected by increased TUNEL staining at 24 h, but not with neuronal necrosis measured by Flouro-Jade. Astrogliosis and microgliosis did not differ, confirming a comparable level of trauma. The gender-specific trait of the secondary inflammatory response may be connected to prostaglandin regulation, which may partially explain gender variances in outcome after TBI.


TBI COX-2 iNOS Sex Gender Secondary inflammation 



This study was funded by ALF Stockholms Läns Landsting and The Swedish Defense.

Conflicts of interest

All authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements) or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.


  1. 1.
    Bains M, Hall ED (2011) Antioxidant therapies in traumatic brain and spinal cord injury. Biochim Biophys Acta 1822(5):675–684CrossRefPubMedCentralPubMedGoogle Scholar
  2. 2.
    Berry C, Ley EJ, Tillou A, Cryer G, Margulies DR, Salim A (2009) The effect of gender on patients with moderate to severe head injuries. J Trauma 67:950–953CrossRefPubMedGoogle Scholar
  3. 3.
    Brait VH, Jackman KA, Walduck AK, Selemidis S, Diep H, Mast AE, Guida E, Broughton BRS, Drummond GR, Sobey CG (2010) Mechanisms contributing to cerebral infarct size after stroke: gender, reperfusion, T lymphocytes, and Nox2-derived superoxide. J Cereb Blood Flow Metab 30:1306–1317CrossRefPubMedCentralPubMedGoogle Scholar
  4. 4.
    Cernak I, O’Connor C, Vink R (2002) Inhibition of cyclooxygenase 2 by nimesulide improves cognitive outcome more than motor outcome following diffuse traumatic brain injury in rats. Exp Brain Res 147:193–199CrossRefPubMedGoogle Scholar
  5. 5.
    Dagerlind A, Friberg K, Bean AJ, Hökfelt T (1992) Sensitive mRNA detection using unfixed tissue: combined radioactive and non-radioactive in situ hybridization histochemistry. Histochemistry 98:39–49CrossRefPubMedGoogle Scholar
  6. 6.
    Eng LF, Ghirnikar RS, Lee YL (2000) Glial fibrillary acidic protein: GFAP-thirty-one years (1969-2000). Neurochem Res 25:1439–1451CrossRefPubMedGoogle Scholar
  7. 7.
    Farace E, Alves WM (2000) Do women fare worse: a metaanalysis of gender differences in traumatic brain injury outcome. J Neurosurg 93:539–545CrossRefPubMedGoogle Scholar
  8. 8.
    Gahm C, Holmin S, Mathiesen T (2000) Temporal profiles and cellular sources of three nitric oxide synthase isoforms in the brain after experimental contusion. Neurosurgery 46:169–177CrossRefPubMedGoogle Scholar
  9. 9.
    Groswasser Z, Cohen M, Keren O (1998) Female TBI patients recover better than males. Brain Inj 12:805–808CrossRefPubMedGoogle Scholar
  10. 10.
    Günther M, Al Nimer F, Gahm C, Piehl F, Mathiesen T (2012) iNOS-mediated secondary inflammatory response differs between rat strains following experimental brain contusion. Acta Neurochir (Wien) 154:689–697CrossRefGoogle Scholar
  11. 11.
    Hein AM, O’Banion MK (2009) Neuroinflammation and memory: the role of prostaglandins. Mol Neurobiol 40:15–32CrossRefPubMedCentralPubMedGoogle Scholar
  12. 12.
    Holmin S, von Gertten C, Sandberg-Nordqvist AC, Lendahl U, Mathiesen T (2001) Induction of astrocytic nestin expression by depolarization in rats. Neurosci Lett 314:151–155CrossRefPubMedGoogle Scholar
  13. 13.
    Injury NCDPoRoPWTB (1999) Consensus conference. rehabilitation of persons with traumatic brain injury. JAMA 282:974–983CrossRefGoogle Scholar
  14. 14.
    Leitgeb J, Mauritz W, Brazinova A, Janciak I, Majdan M, Wilbacher I, Rusnak M (2011) Effects of gender on outcomes after traumatic brain injury. J Trauma 71:1620–1626CrossRefPubMedGoogle Scholar
  15. 15.
    Ley EJ, Short SS, Liou DZ, Singer MB, Mirocha J, Melo N, Bukur M, Salim A (2013) Gender impacts mortality after traumatic brain injury in teenagers. J Trauma Acute Care Surg 75:682–686CrossRefPubMedGoogle Scholar
  16. 16.
    Loane DJ, Byrnes KR (2010) Role of microglia in neurotrauma. Neurotherapeutics 7:366–377CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Lu J, Goh SJ, Tng PY, Deng YY, Ling EA, Moochhala S (2009) Systemic inflammatory response following acute traumatic brain injury. Front Biosci 14:3795–3813CrossRefGoogle Scholar
  18. 18.
    Maghool F, Khaksari M, Khachki AS (2012) Differences in brain edema and intracranial pressure following traumatic brain injury across the estrous cycle: involvement of female sex steroid hormones. Brain Res 1497:61–72CrossRefPubMedGoogle Scholar
  19. 19.
    Manabe Y, Anrather J, Kawano T, Niwa K, Zhou P, Ross ME, Iadecola C (2004) Prostanoids, not reactive oxygen species, mediate COX-2-dependent neurotoxicity. Ann Neurol 55:668–675CrossRefPubMedGoogle Scholar
  20. 20.
    Marcondes FK, Bianchi FJ, Tanno AP (2002) Determination of the estrous cycle phases of rats: some helpful considerations. Braz J Biol 62:609–614CrossRefPubMedGoogle Scholar
  21. 21.
    Marklund N, Bakshi A, Castelbuono DJ, Conte V, McIntosh TK (2006) Evaluation of pharmacological treatment strategies in traumatic brain injury. Curr Pharm Des 12:1645–1680CrossRefPubMedGoogle Scholar
  22. 22.
    Marnett LJ, Wright TL, Crews BC, Tannenbaum SR, Morrow JD (2000) Regulation of prostaglandin biosynthesis by nitric oxide is revealed by targeted deletion of inducible nitric-oxide synthase. J Biol Chem 275:13427–13430CrossRefPubMedGoogle Scholar
  23. 23.
    McCullough LD, Hurn PD (2003) Estrogen and ischemic neuroprotection: an integrated view. Trends Endocrinol Metab 14:228–235CrossRefPubMedGoogle Scholar
  24. 24.
    Miljkovic D, Trajkovic V (2004) Inducible nitric oxide synthase activation by interleukin-17. Cytokine Growth Factor Rev 15:21–32CrossRefPubMedGoogle Scholar
  25. 25.
    Minghetti L (2004) Cyclooxygenase-2 (COX-2) in inflammatory and degenerative brain diseases. J Neuropathol Exp Neurol 63:901–910PubMedGoogle Scholar
  26. 26.
    Mémet S (2006) NF-kappaB functions in the nervous system: from development to disease. Biochem Pharmacol 72:1180–1195CrossRefPubMedGoogle Scholar
  27. 27.
    Ngo ST, Steyn FJ, McCombe PA (2014) Gender differences in autoimmune disease. Front Neuroendocrinol 35:347–369CrossRefPubMedGoogle Scholar
  28. 28.
    O’Connell KM, Littleton-Kearney MT (2013) The role of free radicals in traumatic brain injury. Biol Res Nurs 15:253–263CrossRefPubMedGoogle Scholar
  29. 29.
    Ottochian M, Salim A, Berry C, Chan LS, Wilson MT, Margulies DR (2009) Severe traumatic brain injury: is there a gender difference in mortality? Am J Surg 197:155–158CrossRefPubMedGoogle Scholar
  30. 30.
    Park EM, Cho S, Frys KA, Glickstein SB, Zhou P, Anrather J, Ross ME, Iadecola C (2006) Inducible nitric oxide synthase contributes to gender differences in ischemic brain injury. J Cereb Blood Flow Metab 26:392–401CrossRefPubMedGoogle Scholar
  31. 31.
    Plantman S (2012) Osteopontin is upregulated after mechanical brain injury and stimulates neurite growth from hippocampal neurons through β1 integrin and CD44. Neuroreport 23:647–652CrossRefPubMedGoogle Scholar
  32. 32.
    Plantman S, Ng KC, Lu J, Davidsson J, Risling M (2012) Characterization of a novel rat model of penetrating traumatic brain injury. J Neurotrauma 29:1219–1232CrossRefPubMedGoogle Scholar
  33. 33.
    Renner C, Hummelsheim H, Kopczak A, Steube D, Schneider HJ, Schneider M, Kreitschmann-Andermahr I, Jordan M, Uhl E, Stalla GK (2012) The influence of gender on the injury severity, course and outcome of traumatic brain injury. Brain Inj 26:1360–1371CrossRefPubMedGoogle Scholar
  34. 34.
    Risling M, Plantman S, Angeria M, Rostami E, Bellander BM, Kirkegaard M, Arborelius U, Davidsson J (2011) Mechanisms of blast induced brain injuries, experimental studies in rats. Neuroimage 54(Suppl 1):S89–S97CrossRefPubMedGoogle Scholar
  35. 35.
    Roof RL, Hall ED (2000) Gender differences in acute CNS trauma and stroke: neuroprotective effects of estrogen and progesterone. J Neurotrauma 17:367–388CrossRefPubMedGoogle Scholar
  36. 36.
    Rose JW, Hill KE, Watt HE, Carlson NG (2004) Inflammatory cell expression of cyclooxygenase-2 in the multiple sclerosis lesion. J Neuroimmunol 149:40–49CrossRefPubMedGoogle Scholar
  37. 37.
    Schank JC (2001) Do Norway rats (Rattus norvegicus) synchronize their estrous cycles? Physiol Behav 72:129–139CrossRefPubMedGoogle Scholar
  38. 38.
    Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675CrossRefPubMedGoogle Scholar
  39. 39.
    Si D, Li J, Liu J, Wang X, Wei Z, Tian Q, Wang H, Liu G (2014) Progesterone protects blood–brain barrier function and improves neurological outcome following traumatic brain injury in rats. Exp Ther Med 8:1010–1014PubMedCentralPubMedGoogle Scholar
  40. 40.
    Sköld M, Cullheim S, Hammarberg H, Piehl F, Suneson A, Lake S, Sjögren A, Walum E, Risling M (2000) Induction of VEGF and VEGF receptors in the spinal cord after mechanical spinal injury and prostaglandin administration. Eur J Neurosci 12:3675–3686CrossRefPubMedGoogle Scholar
  41. 41.
    Smith WL, Dewitt DL (1996) Prostaglandin endoperoxide H synthases-1 and -2. Adv Immunol 62:167–215CrossRefPubMedGoogle Scholar
  42. 42.
    von Baumgarten L, Trabold R, Thal S, Back T, Plesnila N (2008) Role of cortical spreading depressions for secondary brain damage after traumatic brain injury in mice. J Cereb Blood Flow Metab 28:1353–1360CrossRefGoogle Scholar
  43. 43.
    Westwood FR (2008) The female rat reproductive cycle: a practical histological guide to staging. Toxicol Pathol 36:375–384CrossRefPubMedGoogle Scholar
  44. 44.
    Woodcock T, Morganti-Kossmann MC (2013) The role of markers of inflammation in traumatic brain injury. Front Neurol 4:18CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Mattias Günther
    • 1
    • 2
  • Stefan Plantman
    • 1
  • Johan Davidsson
    • 3
  • Maria Angéria
    • 1
  • Tiit Mathiesen
    • 4
  • Mårten Risling
    • 1
  1. 1.Department of Neuroscience, Experimental Traumatology UnitKarolinska InstitutetStockholmSweden
  2. 2.Department of Neuroscience, Section of Experimental TraumatologyKarolinska InstitutetStockholmSweden
  3. 3.Division of Vehicle Safety, Department of Applied MechanicsChalmers University of technologyGothenburgSweden
  4. 4.Department of Clinical Neuroscience, Section of NeurosurgeryKarolinska InstitutetStockholmSweden

Personalised recommendations