Comparison of iNOS inhibition by antisense and pharmacological inhibitors after spinal cord injury.
Inducible nitric oxide synthase (iNOS) is a key mediator of inflammation during pathological conditions. We examined, through the use of selective iNOS inhibitors, the role of iNOS in specific pathophysiological processes after spinal cord injury (SCI), including astrogliosis, blood-spinal cord barrier (BSCB) permeability, polymorphonuclear leukocyte infiltration, and neuronal cell death. Administration of iNOS antisense oligonucleotides (ASOs) (intraspinally at 3 h) or the pharmacological inhibitors, N-[3(Aminomethyl) benzyl] acetamidine (1400 W) (i.v./i.p. 3 and 9 h) or aminoguanidine (i.p. at 3 and 9 h) after moderate contusive injury decreased the number of iNOS immunoreactive cells at the injury site by 65.6% (iNOS ASOs), 62.1% (1400 W), or 59% (aminoguanidine) 24 h postinjury. iNOS activity was reduced 81.8% (iNOS ASOs), 56.7% (1400 W), or 67.9% (aminoguanidine) at this time. All iNOS inhibitors reduced the degree of BSCB disruption (plasma leakage of rat immunoglobulins), with iNOS ASO inhibition being more effective (reduced by 58%). Neutrophil accumulation within the injury site was significantly reduced by iNOS ASOs and 1400 W by 78.8% and 20.9%, respectively. Increased astrogliosis was diminished with iNOS ASOs but enhanced following aminoguanidine. Detection of necrotic and apoptotic neuronal cell death by propidium iodide and an FITC-conjugated Annexin V antibody showed that iNOS inhibition could significantly retard neuronal cell death rostral and caudal to the injury site. These novel findings indicate that acute inhibition of iNOS is beneficial in reducing several pathophysiological processes after SCI. Furthermore, we demonstrate that the antisense inhibition of iNOS is more efficacious than currently available pharmacological agents.
Pearse
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DD
The Miami Project to Cure Paralysis and the Department of Neurological Surgery, University of Miami School of Medicine, Miami, Florida 33136, USA.
Chatzipanteli
K
K
Marcillo
A E
AE
Bunge
M B
MB
Dietrich
W D
WD
D000578:Amidines / Q000008:administration & dosage; D000818:Animals; D017302:Annexins / Q000378:metabolism; D001253:Astrocytes / Q000378:metabolism; D001596:Benzylamines / Q000008:administration & dosage; D001812:Blood-Brain Barrier / Q000378:metabolism; D002452:Cell Count; D016923:Cell Death / Q000187:drug effects; D004195:Disease Models, Animal; D004333:Drug Administration Routes / Q000662:veterinary; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006146:Guanidines / Q000008:administration & dosage / Q000494:pharmacology*; D007074:Immunoglobulin G / Q000378:metabolism; D007150:Immunohistochemistry; D007796:Laminectomy / Q000379:methods; D009046:Motor Neurons / Q000187:drug effects* / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism / Q000473:pathology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D009930:Organic Chemicals; D009195:Peroxidase / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011419:Propidium / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013119:Spinal Cord Injuries / Q000188:drug therapy / Q000201:enzymology* / Q000503:physiopathology
C417664:Alexa594; D000578:Amidines; D017302:Annexins; D001596:Benzylamines; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein; D006146:Guanidines; D007074:Immunoglobulin G; C104820:N-(3-(aminomethyl)benzyl)acetamidine; D020319:Oligodeoxyribonucleotides, Antisense; D009930:Organic Chemicals; D011419:Propidium; D009195:Peroxidase; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D010751:Phosphopyruvate Hydratase; C004479:pimagedine