Molecular Biology Reports

, Volume 46, Issue 1, pp 241–250 | Cite as

Allyl isothiocyanate attenuates oxidative stress and inflammation by modulating Nrf2/HO-1 and NF-κB pathways in traumatic brain injury in mice

  • Berrak Caglayan
  • Ertugrul Kilic
  • Arman Dalay
  • Serdar Altunay
  • Mehmet Tuzcu
  • Fusun Erten
  • Cemal Orhan
  • Mehmet Yalcin Gunal
  • Burak Yulug
  • Vijaya Juturu
  • Kazim SahinEmail author
Original Article


Traumatic brain injury (TBI) is the leading cause of mortality and morbidity in young adults and children in the industrialized countries; however, there are presently no FDA approved therapies. TBI results in oxidative stress due to the overproduction of reactive oxygen species and overwhelming of the endogenous antioxidant mechanisms. Recently, it has been reported that antioxidants including phytochemicals have a protective role against oxidative damage and inflammation after TBI. To analyze the effects of a naturally occurring antioxidant molecule, allyl isothiocyanate (AITC), on the nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear factor kappa B (NF-κB) signaling pathways in TBI, a cryogenic injury model was induced in mice. Here, we showed that AITC administered immediately after the injury significantly decreased infarct volume and blood–brain barrier (BBB) permeability. Protein levels of proinflammatory cytokines interleukin-1β (IL1β) and interleukin-6 (IL6), glial fibrillary acidic protein (GFAP) and NF-κB were decreased, while Nrf2, growth-associated protein 43 (GAP43) and neural cell adhesion molecule levels were increased with AITC when compared with vehicle control. Our results demonstrated that the antioxidant molecule AITC, when applied immediately after TBI, provided beneficial effects on inflammatory processes while improving infarct volume and BBB permeability. Increased levels of plasticity markers, as well as an antioxidant gene regulator, Nrf2, by AITC, suggest that future studies are warranted to assess the protective activities of dietary or medicinal AITC in clinical studies.

Graphical abstract


Traumatic brain injury Allyl isothiocyanate NF-κB Nrf2 



Allyl isothiocyanate


Blood–brain barrier


Growth-associated protein 43


Glial fibrillary acidic protein


Intercellular adhesion molecule-1






Neural cell adhesion molecule


Nuclear factor kappa B


Nuclear factor erythroid 2-related factor 2


Reactive oxygen species


Traumatic brain injury



This study was supported by OmniActive Health Technologies Ltd. NJ, USA. This work was also supported in part by The Turkish Academy of Sciences (TUBA) (KS, EK).


This study was supported by OmniActive Health Technologies Inc. (NJ, USA). This work was also supported in part by the Turkish Academy of Sciences.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Dixon KJ (2017) Pathophysiology of traumatic brain injury. Phys Med Rehabil Clin N Am 28(2):215–225Google Scholar
  2. 2.
    Colantonio A, Croxford R, Farooq S, Laporte A, Coyte PC (2009) Trends in hospitalization associated with traumatic brain injury in a publicly insured population, 1992–2002. J Trauma 66:179–183Google Scholar
  3. 3.
    Maas AI, Stocchetti N, Bullock R (2008) Moderate and severe traumatic brain injury in adults. Lancet Neurol 7(8):728–741Google Scholar
  4. 4.
    Feigin VL, Theadom A, Barker-Collo S, Starkey NJ, McPherson K, Kahan M, Dowell A, Brown P, Parag V, Kydd R, Jones K, Jones A, Ameratunga S, BIONIC Study Group (2013) Incidence of traumatic brain injury in New Zealand: a population-based study. Lancet Neurol 12:53–64Google Scholar
  5. 5.
    The Lancet Neurology (2010) Traumatic brain injury: time to end the silence. Lancet Neurol 9:331Google Scholar
  6. 6.
    Andriessen TM, Jacobs B, Vos PE (2010) Clinical characteristics and pathophysiological mechanisms of focal and diffuse traumatic brain injury. J Cell Mol Med 14:2381–2392Google Scholar
  7. 7.
    Winkler EA, Minter D, Yue JK, Manley GT (2016) Cerebral edema in traumatic brain injury: pathophysiology and prospective therapeutic targets. Neurosurg Clin N Am 27(4):473–488Google Scholar
  8. 8.
    Abdul-Muneer PM, Chandra N, Haorah J (2015) Interactions of oxidative stress and neurovascular inflammation in the pathogenesis of traumatic brain injury. Mol Neurobiol 51(3):966–979Google Scholar
  9. 9.
    Hergenroeder GW, Redell JB, Moore AN, Dash PK (2008) Biomarkers in the clinical diagnosis and management of traumatic brain injury. Mol Diagn Ther 12(6):345–358Google Scholar
  10. 10.
    Conaway CC, Yang YM, Chung FL (2002) Isothiocyanates as cancer chemopreventive agents: their biological activities and metabolism in rodents and humans. Curr Drug Metab 3:233–255Google Scholar
  11. 11.
    Xiao D, Srivastava SK, Lew KL, Zeng Y, Hershberger P, Johnson CS, Trump DL, Singh SV (2003) Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits proliferation of human prostate cancer cells by causing G2/M arrest and inducing apoptosis. Carcinogenesis 24(5):891–897Google Scholar
  12. 12.
    Tang L, Zhang Y (2004) Dietary isothiocyanates inhibit the growth of human bladder carcinoma cells. J Nutr 134(8):2004–2010Google Scholar
  13. 13.
    Smith T, Musk SRR, Johnson IT (1996) Allyl isothiocyanate selectively kills undifferentiated HT29 cells in vitro and suppresses aberrant crypt foci in the colonic mucosa of rats. Biochem Soc Trans 24(3):381SGoogle Scholar
  14. 14.
    Smith TK, Lund EK, Parker ML, Clarke RG, Johnson IT (2004) Allyl-isothiocyanate causes mitotic block, loss of cell adhesion and disrupted cytoskeletal structure in HT29 cells. Carcinogenesis 25(8):1409–1415Google Scholar
  15. 15.
    Ernst IM, Wagner AE, Schuemann C, Storm N, Höppner W, Doring F, Stocker A, Rimbach G (2011) Allyl-, butyl- and phenylethyl-isothiocyanate activate Nrf2 in cultured fibroblasts. Pharmacol Res 63(3):233–240Google Scholar
  16. 16.
    Nguyen T, Nioi P, Pickett CB (2009) The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 284(20):13291–13295Google Scholar
  17. 17.
    Yehuda H, Soroka Y, Zlotkin-Frušić M, Gilhar A, Milner Y, Tamir S (2012) Isothiocyanates inhibit psoriasis-related proinflammatory factors in human skin. Inflamm Res 61(7):735–742Google Scholar
  18. 18.
    Lafrenaye AD, Todani M, Walker SA, Povlishock JT (2015) Microglia processes associate with diffusely injured axons following mild traumatic brain injury in the micro pig. J Neuroinflamm 12:186Google Scholar
  19. 19.
    Bryant CD, Zhang NN, Sokoloff G, Fanselow MS, Ennes HS, Palmer AA, McRoberts JA (2008) Behavioral differences among C57BL/6 substrains: implications for transgenic and knockout studies. J Neurogenet 22(4):315–331Google Scholar
  20. 20.
    Jennings M, Batchelor GR, Brain PF, Dick A, Elliott H, Francis RJ, Hubrecht RC, Hurst JL, Morton DB, Peters AG (1998) Refining rodent husbandry: the mouse. report of the rodent refinement working party. Lab Anim 32:233–259Google Scholar
  21. 21.
    Peirson SN, Brown LA, Pothecary CA, Benson LA, Fisk AS (2018) Light and the laboratory mouse. J Neurosci Methods 300:26–36Google Scholar
  22. 22.
    Kelestemur T, Yulug B, Caglayan AB, Beker MC, Kilic U, Caglayan B, Yalcin E, Gundogdu RZ, Kilic E (2016) Targeting different pathophysiological events after traumatic brain injury in mice: role of melatonin and memantine. Neurosci Lett 612:92–97Google Scholar
  23. 23.
    Trevisan G, Rossato MF, Hoffmeister C, Oliveira SM, Silva CR, Matheus FC, Mello GC, Antunes E, Prediger RD, Ferreira J (2013) Mechanisms involved in abdominal nociception induced by either TRPV1 or TRPA1 stimulation of rat peritoneum. Eur J Pharmacol 714(1–3):332–344Google Scholar
  24. 24.
    Beker MC, Caglayan AB, Kelestemur T, Caglayan B, Yalcin E, Yulug B, Kilic U, Hermann DM, Kilic E (2015) Effects of normobaric oxygen and melatonin on reperfusion injury: role of cerebral microcirculation. Oncotarget 6(31):30604–30614Google Scholar
  25. 25.
    Beker MC, Caglayan B, Yalcin E, Caglayan AB, Turkseven S, Gurel B, Kelestemur T, Sertel E, Sahin Z, Kutlu S, Kilic U, Baykal AT, Kilic E (2018) Time-of-day dependent neuronal injury after ischemic stroke: implication of circadian clock transcriptional factor Bmal1 and survival kinase AKT. Mol Neurobiol 55(3):2565–2576Google Scholar
  26. 26.
    Caglayan B, Caglayan AB, Beker MC, Yalcin E, Beker M, Kelestemur T, Sertel E, Ozturk G, Kilic U, Sahin F, Kilic E (2017) Evidence that activation of P2 × 7R does not exacerbate neuronal death after optic nerve transection and focal cerebral ischemia in mice. Exp Neurol 296:23–31Google Scholar
  27. 27.
    Hulsebosch CE, DeWitt DS, Jenkins LW, Prough DS (1998) Traumatic brain injury in rats results in increased expression of Gap-43 that correlates with behavioral recovery. Neurosci Lett 255(2):83–86Google Scholar
  28. 28.
    Boutin C, Schmitz B, Cremer H, Diestel S (2009) NCAM expression induces neurogenesis in vivo. Eur J Neurosci 30(7):1209–1218Google Scholar
  29. 29.
    Washington PM, Villapol S, Burns MP (2016) Polypathology and dementia after brain trauma: does brain injury trigger distinct neurodegenerative diseases, or should they be classified together as traumatic encephalopathy? Exp Neurol 3:381–388Google Scholar
  30. 30.
    Venegoni W, Shen Q, Thimmesch AR, Bell M, Hiebert JB, Pierce JD (2017) The use of antioxidants in the treatment of traumatic brain injury. J Adv Nurs 73(6):1331–1338Google Scholar
  31. 31.
    Abdul-Muneer PM, Schuetz H, Wang F, Skotak M, Jones J, Gorantla S, Zimmerman MC, Chandra N, Haorah J (2013) Induction of oxidative and nitrosative damage leads to cerebrovascular inflammation in an animal model of mild traumatic brain injury induced by primary blast. Free Radic Biol Med 60:282–291Google Scholar
  32. 32.
    de Roos B, Duthie GG (2015) Role of dietary pro-oxidants in the maintenance of health and resilience to oxidative stress. Mol Nutr Food Res 59(7):1229–1248Google Scholar
  33. 33.
    Albert-Weissenberger C, Sirén AL (2010) Experimental traumatic brain injury. Exp Transl Stroke Med 2:16Google Scholar
  34. 34.
    Kamm K, Vanderkolk W, Lawrence C, Jonker M, Davis AT (2006) The effect of traumatic brain injury upon the concentration and expression of interleukin-1beta and interleukin-10 in the rat. J Trauma 60:152–157Google Scholar
  35. 35.
    Ley EJ, Clond MA, Singer MB, Shouhed D, Salim A (2011) IL6 deficiency affects function after traumatic brain injury. J Surg Res 170(2):253–256Google Scholar
  36. 36.
    Holmin S, Mathiesen T (2000) Intracerebral administration of interleukin-1beta and induction of inflammation, apoptosis, and vasogenic edema. J Neurosurg 92:108–120Google Scholar
  37. 37.
    Quagliarello VJ, Wispelwey B, Long WJ Jr, Scheld WM (1991) Recombinant human interleukin-1 induces meningitis and blood-brain barrier injury in the rat. Characterization and comparison with tumor necrosis factor. J Clin Invest 87(4):1360–1366Google Scholar
  38. 38.
    Bevilacqua MP, Pober JS, Wheeler ME, Cotran RS, Gimbrone MA Jr (1985) Interleukin-1 activation of vascular endothelium. Effects on procoagulant activity and leukocyte adhesion. Am J Pathol 121(3):394–403Google Scholar
  39. 39.
    Frugier T, Morganti-Kossmann MC, O’Reilly D, McLean CA (2010) In situ detection of inflammatory mediators in post mortem human brain tissue after traumatic injury. J Neurotrauma 27:497–507Google Scholar
  40. 40.
    Woodcock T, Morganti-Kossmann MC (2013) The role of markers of inflammation in traumatic brain injury. Front Neurol 4:4–18Google Scholar
  41. 41.
    Vos PE (2011) Biomarkers of focal and diffuse traumatic brain injury. Crit Care 15(4):183Google Scholar
  42. 42.
    Nylén K, Ost M, Csajbok LZ, Nilsson I, Blennow K, Nellgård B, Rosengren L (2006) Increased serum-GFAP in patients with severe traumatic brain injury is related to outcome. J Neurol Sci 240(1–2):85–91Google Scholar
  43. 43.
    Kaltschmidt B, Widera D, Kaltschmidt C (2005) Signaling via NF-kappaB in the nervous system. Biochim Biophys Acta 1745:287–299Google Scholar
  44. 44.
    Laird MD, Vender JR, Dhandapani KM (2008) Opposing roles for reactive astrocytes following traumatic brain injury. Neurosignals 16:154–164Google Scholar
  45. 45.
    Salminen A, Liu PK, Hsu CY (1995) Alteration of transcription factor binding activities in the ischemic rat brain. Biochem Biophys Res Commun 212:939–944Google Scholar
  46. 46.
    Nonaka M, Chen XH, Pierce JE, Leoni MJ, McIntosh TK, Wolf JA, Smith DH (1999) Prolonged activation of NF-kappaB following traumatic brain injury in rats. J Neurotrauma 16(11):1023–1034Google Scholar
  47. 47.
    Xiao G, Wei J (2005) Effects of β-Aescin on the expression of nuclear factor-κB and tumor necrosis factor-α after traumatic brain injury in rats. J Zhejiang Univ Sci B 6(1):28–32Google Scholar
  48. 48.
    Wardyn JD, Ponsford AH, Sanderson CM (2015) Dissecting molecular cross-talk between Nrf2 and NF-κB response pathways. Biochem Soc Trans 43(4):621–626Google Scholar
  49. 49.
    Jakubíková J, Sedlák J, Bod’o J, Bao Y (2006) Effect of isothiocyanates on nuclear accumulation of NF-kappaB, Nrf2, and thioredoxin in caco-2 cells. J Agric Food Chem 54(5):1656–1662Google Scholar
  50. 50.
    Wagner AE, Boesch-Saadatmandi C, Dose J, Schultheiss G, Rimbach G (2012) Anti-inflammatory potential of allyl-isothiocyanate—role of Nrf2, NF-(κ) B and microRNA-155. J Cell Mol Med 16(4):836–843Google Scholar
  51. 51.
    Liddell JR (2017) Interplay between Nrf2 and NF-κB in neuroinflammatory diseases. J Clin Cell Immunol 8:489. Google Scholar
  52. 52.
    Whalen MJ, Carlos TM, Dixon CE, Schiding JK, Clark RS, Baum E, Yan HQ, Marion DW, Kochanek PM (1999) Effect of traumatic brain injury in mice deficient in intercellular adhesion molecule-1: assessment of histopathologic and functional outcome. J Neurotrauma 16:299–309Google Scholar
  53. 53.
    Bowes MP, Zivin JA, Rothlein R (1993) Monoclonal antibody to the ICAM-1 adhesion site reduces neurological damage in a rabbit cerebral embolism stroke model. Exp Neurol 119:215–219Google Scholar
  54. 54.
    Knoblach SM, Faden AI (2002) Administration of either anti-intercellular adhesion molecule-1 or a nonspecific control antibody improves recovery after traumatic brain injury in the rat. J Neurotrauma 19:1039–1050Google Scholar
  55. 55.
    You WC, Wang CX, Pan YX, Zhang X, Zhou XM, Zhang XS, Shi JX, Zhou ML (2013) Activation of nuclear factor-κB in the brain after experimental subarachnoid hemorrhage and its potential role in delayed brain injury. PLoS ONE 8(3):e60290Google Scholar
  56. 56.
    Gorup D, Bohaček I, Miličević T, Pochet R, Mitrečić D, Križ J, Gajović S (2015) Increased expression and colocalization of GAP43 and CASP3 after brain ischemic lesion in mouse. Neurosci Lett 597:176–182Google Scholar
  57. 57.
    Bautista DM, Jordt SE, Nikai T, Tsuruda PR, Read AJ, Poblete J, Yamoah EN, Basbaum AI, Julius D (2006) TRPA1 mediates the inflammatory actions of environmental irritants and proalgesic agents. Cell 124(6):1269–1282Google Scholar
  58. 58.
    Bautista DM, Pellegrino M, Tsunozaki M (2013) TRPA1: a gatekeeper of inflammation. Annu Rev Physiol 75:181–200Google Scholar
  59. 59.
    Corrigan F, Mander KA, Leonard AV, Vink R (2016) Neurogenic inflammation after traumatic brain injury and its potentiation of classical inflammation. J Neuroinflamm 13(1):264Google Scholar
  60. 60.
    Sun J, Ramnath RD, Zhi L, Tamizhselvi R, Bhatia M (2008) Substance P enhances NF-kappaB transactivation and chemokine response in murine macrophages via ERK1/2 and p38 MAPK signaling pathways. Am J Physiol Cell Physiol 294(6):C1586–C1596Google Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Medical Biology, International School of MedicineIstanbul Medipol UniversityIstanbulTurkey
  2. 2.Department of PhysiologyIstanbul Medipol UniversityIstanbulTurkey
  3. 3.Regenerative and Restorative Medical Research CenterIstanbul Medipol UniversityIstanbulTurkey
  4. 4.Division of Biology, Faculty of ScienceFirat UniversityElazigTurkey
  5. 5.Animal Nutrition Department, School of Veterinary MedicineFirat UniversityElazigTurkey
  6. 6.Department of Neurology, School of MedicineIstanbul Medipol UniversityIstanbulTurkey
  7. 7.Research and DevelopmentOmniActive Health Technologies Inc.MorristownUSA

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