Acta Neurochirurgica

, Volume 129, Issue 1–2, pp 58–63

Neuropathological endpoints in experimental stroke pharmacotherapy: The importance of both early and late evaluation

  • J. Valtysson
  • L. Hillered
  • P. Andiné
  • H. Hagberg
  • L. Persson
Experimental Research

Summary

This study adresses the issue of endpoint selection in the evaluation of neuroprotective drugs in experimental focal ischaemia. Previous work with the permanent middle cerebral artery (MCA) occlusion model in the rat has demonstrated that the ischaemic lesion does not acquire its final appearance until at least 28 days after the ictus. Therefore, the effect of the NMDA receptor blocker MK-801 (dizocilpine maleate) was evaluated both early (3 days) and late (28 days) after MCA occlusion to determine if the previously reported protective effect of a single post-ischaemic dose of MK801 found in acute experiments remained after 28 days.

Mk-801 (0.5mg/kg, i.v.) or isotonic saline was randomly given to rats 30 min after MCA occlusion. Infarct volume and volume of ipsilateral and contralateral hemispheres were estimated from camera lucida drawings of 8 defined coronal histological sections of the brain. As expected, a 40% (p<0.05) reduction of infarct size was found in MK-801 treated rats after 3 days. In animals evaluated 28 days after MCA occlusion, no significant difference in infarct size, total tissue loss (infarct volume + ipsilateral hemisphere atrophy) or remaining non-infarcted tissue (contralateral hemisphere — total tissue loss) was seen between the MK-801 and placebo treated rats.

The results suggest that the single dose treatment with MK-801 postponed the evolution of the infarct, which at 3 days after MCA occlusion is still in progress, possibly by ameliorating oedema formation. It remains to be shown if a multiple dose treatment with NMDA receptor antagonists improves the final neuropathological outcome after experimental stroke. The study illustrates the importance of including a late endpoint when evaluating the efficacy of neuroprotective stroke therapy.

Keywords

Focal cerebral ischaemia MK-801 infarct size neuroprotection 

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References

  1. 1.
    Tamura A, Graham DI, McCulloch J, Teasdale TM (1981) Focal cerebral ischemia in the rat. 1. Description of technique and early neuropathological consequences following middle cerebral artery occlusion. J Cereb Blood Flow Metab 1: 53–60PubMedGoogle Scholar
  2. 2.
    Persson L, Hårdemark H-G, Bolander H, Hillered L, Olsson Y (1989) Neurologic and neuropathologic outcome after middle cerebral artery occlusion in rats. Stroke 20: 641–645PubMedGoogle Scholar
  3. 3.
    Osborne KA, Shigeno T, Balarsky AM, Ford I, McCulloch J, Teasdale GM, Graham DI (1987) Quantitative assessment of early brain damage in a rat model of focal cerebral ischaemia. J Neurosurg Psychiatry 50: 402–410Google Scholar
  4. 4.
    Kirino T, Tamura A, Sano K (1984) Delayed neuronal death in the rat hippocampus following transient forebrain ischemia. Acta Neuropathol (Berl) 64: 139–147Google Scholar
  5. 5.
    Ginsberg MD, Busto R (1989) Rodent models of cerebral ischemia. Stroke 20: 1627–1642PubMedGoogle Scholar
  6. 6.
    Albers GW, Goldberg MP, Choi DW (1989) N-methyl-D-aspartate antagonists: ready for clinical trial in brain ischemia? Ann Neurol 25: 398–403PubMedGoogle Scholar
  7. 7.
    Scatton B, Carter C, Benavidès J, Giroux C (1991) N-methyl-D-aspartate receptor antagonists: a novel therapeutic perspective for the treatment of ischemic brain injury. Cerebrovasc Dis 1: 21–135Google Scholar
  8. 8.
    Bolander HG, Persson L, Hillered L, d'Argy R, Pontén U, Olsson Y (1989) Regional cerebral blood flow changes after middle cerebral artery occlusion in rats. Stroke 20: 930–937PubMedGoogle Scholar
  9. 9.
    Ozyurt E, Graham DI, Woodruff GN, McCulloch J (1988) Protective effect of the glutamate antagonist, MK-801 in focal cerebral ischemia in the cat. J Cereb Blood Flow Metab 8: 138–143PubMedGoogle Scholar
  10. 10.
    Park CK, Nehls DG, Graham DI, Teasdale GM, McCulloch J (1988a) The glutamate antagonist MK-801 reduces focal ischemic brain damage in the rat. Ann Neurol 24: 543–551PubMedGoogle Scholar
  11. 11.
    Park CK, Nehls DG, Graham DI, Teasdale GM, McCulloch J (1988b) Focal cerebral ischemia in the cat: treatment with the glutamate antagonist MK-801 after induction of ischemia. J Cereb Blood Flow Metab 8: 757–762PubMedGoogle Scholar
  12. 12.
    Gotti B, Benavidès J, MacKenzie ET, Scatton B (1990) The pharmacotherapy of focal cortical ischemia in the mouse. Brain Res 522: 290–307PubMedGoogle Scholar
  13. 13.
    Bielenberg GW, Beck T (1991) The effects of dizocilpine (MK-801), phencyclidine and nimodipine on infarct size 48 h after middle cerebral artery occlusion in the rat. Brain Res 552: 338–342PubMedGoogle Scholar
  14. 14.
    Gill R, Andiné P, Hillered L, Persson L, Hagberg H (1992) The effect of MK-801 on cortical spreading depression in the penumbral zone following focal ischemia in the rat. J Cereb Blood Flow Metab 12: 371–379PubMedGoogle Scholar
  15. 15.
    Valtysson J, Hillered L, Andiné P, Hagberg H, Persson L (1991) Long-term effects of MK-801 on the size of the ischemic lesion in MCA occluded rats. J Cereb Blood Flow Metab 11 [Suppl 2]: 284Google Scholar
  16. 16.
    Persson L, Valtysson J, Andiné P, Hagberg H, Hillered L (1992) Neuropathologic end points in experimental stroke therapy (MK-801): the importance of both early and late evaluation. Soc Neurosci Abstr 18: 1452Google Scholar
  17. 17.
    Hillered L, Kotwica Z, Ungerstedt U (1991) Interstitial and cerebrospinal fluid levels of energy-related metabolites after middle cerebral artery occlusion in rats. Res Exp Med 191: 219–225Google Scholar
  18. 18.
    Bederson JB, Pitts LH, Tsuji M, Nishimura MC, Davis, Barktowski H (1986) Rat middle cerebral artery occlusion: evaluation of the model and development of a neurological examination. Stroke 17: 472–476PubMedGoogle Scholar
  19. 19.
    Hårdemark H-G, Persson L, Bolander H, Hillered L, Olsson Y, Påhlman S (1988) Neuron-specific enolase is a marker of cerebral ischemia and infarct size in rat cerebrospinal fluid. Stroke 19: 1140–1144PubMedGoogle Scholar
  20. 20.
    Blackwood W (1969) Vascular disease of the central nervous system. In: Blackwood W, McMenemey HW, Meyer A, Norman RM, Russel DS (eds) Greenfield's neuropathology. Edward Arnold, London, pp 71–137Google Scholar
  21. 21.
    Graham DI (1985) The pathology of brain ischemia and possibilities for therapeutic intervention. Br J Anaesth 57: 3–17PubMedGoogle Scholar
  22. 22.
    Katzman R, Clasen R, Klatzo I, Stirling Meyer J, Pappius HM, Waltz AG (1977) Brain edema in stroke. Stroke 8: 510–540Google Scholar
  23. 23.
    Thoumas K-å, Kotwica Z, Bergström K, Bolander HG, Hillered L, Olsson Y, Pontén U, Persson L (1991) MRI of cerebral ischaemia in rats with occlusion of the middle cerebral artery. Eur Radiol 1: 118–123Google Scholar
  24. 24.
    Nedergaard M, Astrup J (1986) Infarct rim: effect of hyperglycemia on direct current potential and [14C]2-deoxyglucose phosphorylation. J Cereb Blood Flow Metab 6: 607–615PubMedGoogle Scholar
  25. 25.
    Hillered L, Hallström å, SegersvÄrd S, Persson L, Ungerstedt U (1989) Dynamics of extracellular metabolites in the striatum after middle cerebral artery occlusion in the rat monitored by intracerebral microdialysis. J Cereb Blood Flow Metab 9: 607–616PubMedGoogle Scholar
  26. 26.
    Butcher SP, Bullock R, Graham DI, McCulloch J (1990) Correlation between amino acid release and neuropathologic outcome in rat brain following middle cerebral artery occlusion. Stroke 21: 1727–1733PubMedGoogle Scholar
  27. 27.
    Hansen AJ (1985) Effect of anoxia on ion distribution in the brain. Physiol Rev 65: 101–148PubMedGoogle Scholar
  28. 28.
    Iijima T, Mies G, Hossmann K-A (1992) Repeated negative DC deflections in rat cortex following middle cerebral artery occlusion are abolished by MK-801: effect on volume of ischemic injury. J Cereb Blood Flow Metab 12: 727–733PubMedGoogle Scholar
  29. 29.
    Swanson RA, Morton MT, Wu GT, Savalos R, Davidson C, Sharp FR (1990) A semiautomated method for measuring brain infarct volume. J Cereb Blood Flow Metab 10: 290–293PubMedGoogle Scholar
  30. 30.
    Lin T-N, He YY, Wu G, Khan M, Hsu CY (1993) Effect of brain edema on infarct volume in a focal cerebral ischemia model in rats. Stroke 24: 117–121PubMedGoogle Scholar
  31. 31.
    McIntosh TK, Vink R, Soares H, Hayes R, Simon R (1990) Effect of non-competitive blockade of N-methyl-D-aspartate receptors on the neurochemical sequelae of experimental brain injury. J Neurochem 55: 1170–1179PubMedGoogle Scholar
  32. 32.
    Oh SM, Betz AL (1991) Interaction between free radicals and excitatory amino acids in the formation of ischemic brain edema. Stroke 22: 915–921PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • J. Valtysson
    • 1
  • L. Hillered
    • 1
    • 2
  • P. Andiné
    • 3
  • H. Hagberg
    • 3
    • 4
  • L. Persson
    • 1
  1. 1.Department of NeurosurgeryUppsala University HospitalSweden
  2. 2.Department of Clinical ChemistryUppsala University HospitalSweden
  3. 3.Institute of NeurobiologyUniversity of GöteborgSweden
  4. 4.Department of Obstetrics and GynaecologyUniversity of GöteborgSweden

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