Role of endothelial nitric oxide generation and peroxynitrite formation in reperfusion injury after focal cerebral ischemia.
OBJECTIVE
Reperfusion injury is one of the factors that unfavorably affects stroke outcome and shortens the window of opportunity for thrombolysis. Surges of nitric oxide (NO) and superoxide generation on reperfusion have been demonstrated. Concomitant generation of these radicals can lead to formation of the strong oxidant peroxynitrite during reperfusion.


METHODS
We have examined the role of NO generation and peroxynitrite formation on reperfusion injury in a mouse model of middle cerebral artery occlusion (2 hours) and reperfusion (22 hours). The infarct volume was assessed by 2,3,5-triphenyl tetrazolium chloride staining; blood-brain barrier permeability was evaluated by Evans blue extravasation. Nitrotyrosine formation and matrix metalloproteinase-9 expression were detected by immunohistochemistry.


RESULTS
Infarct volume was significantly decreased (47%) in animals treated with the nonselective nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L-arginine (L-NA) at reperfusion. The specific inhibitor of neuronal NOS, 7-nitroindazole (7-NI), given at reperfusion, showed no protection, although preischemic treatment with 7-NI decreased infarct volume by 40%. Interestingly, prereperfusion administration of both NOS inhibitors decreased tyrosine nitration (a marker of peroxynitrite toxicity) in the ischemic area. L-NA treatment also significantly reduced vascular damage, as indicated by decreased Evans blue extravasation and matrix metalloproteinase-9 expression.


CONCLUSIONS
These data support the hypothesis that in addition to the detrimental action of NO formed by neuronal NOS during ischemia, NO generation at reperfusion plays a significant role in reperfusion injury, possibly through peroxynitrite formation. Contrary to L-NA, failure of 7-NI to protect against reperfusion injury suggests that the source of NO is the cerebrovascular compartment.
Gürsoy-Ozdemir
Y
Y
Department of Neurology, Faculty of Medicine, and Institute of Neurological Sciences and Psychiatry, Hacettepe University, Ankara, Turkey.
Bolay
H
H
Saribaş
O
O
Dalkara
T
T
D000818:Animals; D015415:Biomarkers; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology; D004396:Coloring Agents; D004730:Endothelium, Vascular / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005070:Evans Blue; D007191:Indazoles / Q000494:pharmacology; D020244:Infarction, Middle Cerebral Artery / Q000150:complications* / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis; D051379:Mice; D009566:Nitrates / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D019335:Nitroarginine / Q000494:pharmacology; D016877:Oxidants / Q000378:metabolism*; D010539:Permeability / Q000187:drug effects; D015427:Reperfusion Injury / Q000188:drug therapy / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000096:biosynthesis
D015415:Biomarkers; D004396:Coloring Agents; D004791:Enzyme Inhibitors; D007191:Indazoles; D009566:Nitrates; D016877:Oxidants; D019335:Nitroarginine; C039642:peroxynitric acid; D009569:Nitric Oxide; C002744:3-nitrotyrosine; D014443:Tyrosine; D005070:Evans Blue; D019001:Nitric Oxide Synthase; D020780:Matrix Metalloproteinase 9; C080122:7-nitroindazole