Inducible nitric oxide synthase: a possible key factor in the pathogenesis of chronic vasospasm after experimental subarachnoid hemorrhage.
OBJECTIVE
The role of nitric oxide (NO) in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH) is not well understood. Nitric oxide is a well-established vasodilatory substance; however, in SAH, NO may become a major source for the production of injurious free-radical species, leading to chronic cerebral vasospasm. Reactive overproduction of NO to counteract vascular narrowing might potentiate the detrimental effects of NO. The focus of the present study is to determine the extent of reactive induction of inducible nitric oxide synthase (iNOS) after experimental SAH.


METHODS
Chronic vasospasm was induced in male Wistar rats by an injection of autologous blood (100 microl) into the cisterna magna followed by a second injection 24 hours later. A control group of 10 animals was treated with injections of 0.9% sodium chloride solution. Vasospasm was verified by pressure-controlled angiography after retrograde cannulation of the external carotid artery 7 days later. In 11 of 15 animals radiographic evidence of cerebral vasospasm was seen. The animals were perfusion fixed and their brains were removed for immunohistochemical assessment. With the aid of a microscope, staining for iNOS was quantified in 40-microm floating coronal sections. Immunohistochemical staining for iNOS was markedly more intense in animals with significant angiographic evidence of vasospasm. Virtually no staining was observed in control animals. Seven days after the second experimental SAH, labeling of iNOS was found in endothelial cells, in vascular smooth-muscle cells, and, above all, in adventitial cells. Some immunohistochemical staining of iNOS was observed in rod cells (activated microglia), in glial networks, and in neurons.


CONCLUSIONS
The present study demonstrates induction of iNOS after experimental SAH.
Widenka
D C
DC
Department of Neurosurgery and Institute of Neuropathology, Klinikum Grosshadern, Ludwig-Maximilians University Munich, Germany.
Medele
R J
RJ
Stummer
W
W
Bise
K
K
Steiger
H J
HJ
D000818:Animals; D001158:Arteries / Q000201:enzymology; D001921:Brain / Q000201:enzymology; D002533:Cerebral Angiography; D002560:Cerebrovascular Circulation / Q000502:physiology; D002908:Chronic Disease; D007150:Immunohistochemistry / Q000379:methods; D002546:Ischemic Attack, Transient / Q000000981:diagnostic imaging / Q000201:enzymology / Q000209:etiology* / Q000473:pathology; D008297:Male; D019001:Nitric Oxide Synthase / Q000378:metabolism / Q000502:physiology*; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D051381:Rats; D017208:Rats, Wistar; D013194:Staining and Labeling; D013345:Subarachnoid Hemorrhage / Q000150:complications* / Q000201:enzymology
D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496318:Nos2 protein, rat; C496316:Nos3 protein, rat