Molecular and Cellular Biochemistry

, Volume 303, Issue 1–2, pp 19–25 | Cite as

Reactive oxygen species plasmatic levels in ischemic stroke

  • Laura Nanetti
  • Ruja Taffi
  • Arianna Vignini
  • Cinzia Moroni
  • Francesca Raffaelli
  • Tiziana Bacchetti
  • Mauro Silvestrini
  • Leandro Provinciali
  • Laura Mazzanti


Oxidative stress is probably one of the mechanisms involved in neuronal damage induced by ischemia-reperfusion, and the antioxidant activity of plasma may be an important factor providing protection from neurological damage caused by stroke-associated oxidative stress. The aim of this study was to investigate the status of oxidative stress, NO and ONOO levels in patients with atherothrombotic and lacunar acute ischemic stroke and iNOS, eNOS and nitrotyrosine expression in the same patients. Plasma ONOO levels were significantly higher in patients than in controls while NO decreases in patients in respect to controls. Densitometric analysis of bands indicated that iNOS and N-Tyr protein levels were significantly higher in patients in respect to controls. This study has highlighted a significant NO decrease in our patients compared with controls and this is most probably due to the increased expression of inducible NO synthase by the effect of thrombotic attack. In fact, the constitutive NO isoforms, which produce small amounts of NO, are beneficial, while activation of the inducible isoform of NO, which produces much more NO, causes injury, being its toxicity greatly enhanced by generation of peroxynitrite. The significant ONOO increase observed in our patients, compared to controls, is most probably due to reaction of NO with O 2 ·− . These findings suggest that free radical production and oxidative stress in ischemic stroke might have a major role in the pathogenesis of ischemic brain injury. Peroxynitrite might be the main marker of brain damage and neurological impairment in acute ischemic stroke.


Nitric oxide Peroxynitrite Nitric oxide synthase Plasma Stroke 



The present work was supported by a grant Fondazione Cavaliere del Lavoro Mario Magnetto to L M.


  1. 1.
    Murray CJ, Lopez AD (1997) Mortality by cause for eight regions of the word: global burden of disease study. Lancet 349:1269–1276PubMedCrossRefGoogle Scholar
  2. 2.
    Toni D, Fiorelli M, Gentile M, Bastianello S, Sacchetti ML, Argentino C, Pozzilli C, Fieschi C (1995) Progressing neurological deficit secondary to acute ischemic stroke: a study on predictability, pathogenesis, and prognosis. Arc Neurol 52:670–675Google Scholar
  3. 3.
    Davaloas A, Cendra E, Teruel J, Martinez M, Genis D (1990) Deterioratine ischemic stroke: risk factor and prognosis. Neurology 40:1865–1869Google Scholar
  4. 4.
    Loscalzo J (1995) Nitric oxide and vascular desease. N Engl Y Med 333:251–253CrossRefGoogle Scholar
  5. 5.
    Rudic RD, Seesa WC (1999) Nitric oxide in endhotelial dysfunction and vascular remodeling: clinical correlates and experimental links. Am J Hum Genet 64:673–677PubMedCrossRefGoogle Scholar
  6. 6.
    Marletta MA, Hurshman AR, Rusche KM (1998) Catalysis by nitric oxide synthase. Curr Opin Chem Biol 2(5):656–663PubMedCrossRefGoogle Scholar
  7. 7.
    Nathan C (1997) Nitric oxide as a secretory product of mammalian cells. FASEB J 6:3051–3064Google Scholar
  8. 8.
    Guix FX, Uribesalgo I, Coma M, Munoz FJ (2005) The physiology and pathophysiology of nitric oxide in the brain. Prog Neurobiol 76(2):126–152PubMedCrossRefGoogle Scholar
  9. 9.
    Szabo C (2003) Multiple pathway of peroxynitrite cytotoxicity. Toxicol Lett 140–141:105–112PubMedCrossRefGoogle Scholar
  10. 10.
    Beckman JS, Koppenol WH (1996) Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and ugly. Am J Physiol 271(5 Pt 1):C1424–C1437PubMedGoogle Scholar
  11. 11.
    Kontos HA (1985) Oxygen radicals species in cerebral vascular injury. Cir Res 57:508–516Google Scholar
  12. 12.
    Chan PH (1989) Role of oxidants in ischemia brain damage. Cerebrovasc Brain Metab Rev 1:165–211Google Scholar
  13. 13.
    Borg J, London J (2000) Copper/zinc superoxide dismutase overexpression promotes survival of cortical neurons exposed to neurotoxins in vitro. J Neurosci Res 70:180–189CrossRefGoogle Scholar
  14. 14.
    Mattson MP, Duan W, Pedersen WA, Culmsee C (2001) Neurodegenerative disorders and ischemic brain diseases. Apoptosis 6(1–2):69–81PubMedCrossRefGoogle Scholar
  15. 15.
    Keynes RG, Garthwaite J (2004) Nitric oxide and its role in ischemia brain injury. Curr Mol Med 4(2):179–191PubMedCrossRefGoogle Scholar
  16. 16.
    Kander A, Frazzini VI, Solomon RA, Trifilettti RR (1993) Nitric oxide production durino focal ceebral ischemia in rats. Stroke 24:1709–1716Google Scholar
  17. 17.
    Dawson VL, Dawson TM (1996) Nitric oxide neurotoxicity. J Chem Neuroanat 10:179–190PubMedCrossRefGoogle Scholar
  18. 18.
    Aho K, Harmsen P, Hatano S et al (1980) Cerebrovascular disease in the community: results of a WHO collaborative study. Bull WHO 58:113–130PubMedGoogle Scholar
  19. 19.
    Bamford J, Sandercock P, Dennis M (1991) Classification and natural history of clinically identifiable subtype of cerebral infarction. Lancet 337:1521–1526PubMedCrossRefGoogle Scholar
  20. 20.
    Wityle RJ, Pessin MS, Kaplan RF, Caplan LR (1994) Serial assessment of acute stroke using the NIH stroke scale. Stroke 25:362–365Google Scholar
  21. 21.
    Van Swieten JC, Koudstaal PJ, Visser MC (1988) Interobserver agreement for the assessment of handicap in stroke patients. Stroke 19:1083–1092Google Scholar
  22. 22.
    Chen LY, Mehta JL (1996) Further evidence of the presence of constitutive and inducible nitric oxide synthase isoforms in human platelets. J Cardiovasc Pharmacol 27(1):154–158PubMedCrossRefGoogle Scholar
  23. 23.
    Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteine-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  24. 24.
    Tannous M, Rabini RA, Vignini A, Moretti N, Fumelli P, Zielinski B (1999) Evidence for iNOS-dependent peroxynitrite production in diabetic patients. Diabetologia 42:539–544PubMedCrossRefGoogle Scholar
  25. 25.
    Stefano GB, Ottaviani E (2002) The biochemical substrate of nitric oxide signaling is present in primitive non-cognitive organisms. Brain Res 924(1):82–89Google Scholar
  26. 26.
    Esch T, Stefano GB, Fricchione GL, Benson H (2002) Stress related disease- a potential role for nitric oxide. Med Sci Monit 8(6):103–118Google Scholar
  27. 27.
    Vignini A, Nanetti L, Moroni C, Tanase L, Bartolini M, Luzzi S, Provinciali L, Mazzanti L (2006) Modifications of platelet from Alzheimer disease patients: a possible relation between membrane properties and NO metabolites. Neurobiol Aging (in press)Google Scholar
  28. 28.
    Sies H (1985) Oxidative stress: introductory remarks. In: Sies H (ed) Oxidative stress. Academic Press, London, pp 1–8Google Scholar
  29. 29.
    Sies H (2000) What is oxidative stress? In: Keaney JF Jr (ed) Oxidative stress and vascular disease. Kluwer Academic Publishers, Boston, pp 1–8Google Scholar
  30. 30.
    Sies H (1997) Antioxidants in disease mechanisms and therapy. Academic Press, San DiegoGoogle Scholar
  31. 31.
    Lebel CP, Bondy SC (1991) Oxygen radicals: a common mediators of neurotoxicity. Neurotoxicol Teratol 13:341–346PubMedCrossRefGoogle Scholar
  32. 32.
    Polidori MC, Frei B, Cherubini A, Nelles G, Rordorf G, Keaney JF Jr, Schwamm L, Mecocci P, Koroshetz WJ, Beal MF (1998) Increased plasma levels of lipid hydroperoxides in patients with ischemic stroke. Free Radic Biol Med 25:561–567PubMedCrossRefGoogle Scholar
  33. 33.
    Stocker A, Frei B (1991) Endogenous antioxidant defenses in human blood plasma. In: Sies H (ed) Oxidative stress: oxidants and antioxidants. Academic Press, London, pp 213–243Google Scholar
  34. 34.
    Ischiropoulos H, Zhu L, Chen J, Tsai M, Martin JC, Smith CD (1992) Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase. Arch Biochem Biophys 298:431–437PubMedCrossRefGoogle Scholar
  35. 35.
    Gruener N, Gross B, Gozlan O, Barak M (1994) Increase in superoxide dismutase after cerebrovascular accident. Life Sci 54(11):711–713PubMedCrossRefGoogle Scholar
  36. 36.
    Li Y, Wang X, Yang Z (1995) Neuron-specific enolase in patients with acute ischemic stroke and related dementia. Chin Med J (Engl) 108(3):221–223Google Scholar
  37. 37.
    Yoshida E, Mokuno K, Aoki S, Takahashi A, Riku S, Murayama T, Yanagi T, Kato K (1994) Cerebrospinal fluid levels of superoxide dismutases in neurological diseases detected by sensitive enzyme immunoassays. J Neurol Sci 124(1):25–31PubMedCrossRefGoogle Scholar
  38. 38.
    Kasina S, Wasia R, Fasim A, Radhika KV, Singh SS (2006) Phorbol ester mediated activation of inducible nitric oxide synthase results in platelet profilin nitration. Nitric Oxide 14(1):65–71PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Laura Nanetti
    • 1
  • Ruja Taffi
    • 2
  • Arianna Vignini
    • 1
  • Cinzia Moroni
    • 1
  • Francesca Raffaelli
    • 1
  • Tiziana Bacchetti
    • 1
  • Mauro Silvestrini
    • 2
  • Leandro Provinciali
    • 2
  • Laura Mazzanti
    • 1
  1. 1.Institute of BiochemistryUniversità Politecnica delle MarcheAnconaItaly
  2. 2.Department of Neurological ScienceUniversità Politecnica delle MarcheAnconaItaly

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