Hyperbaric oxygen reduces tissue hypoxia and hypoxia-inducible factor-1 alpha expression in focal cerebral ischemia.
OBJECTIVE
The usefulness of hyperbaric oxygen (HBO) and normobaric hyperoxia in acute ischemic stroke is being reexplored because both improve outcome in experimental cerebral ischemia. However, even the basic mechanisms underlying oxygen therapy are poorly understood. We investigated the effect of both oxygen therapies on tissue hypoxia and on the transcription factor hypoxia-inducible factor-1 alpha.


METHODS
Mice were subjected to filament-induced middle cerebral artery occlusion for 2 hours. Twenty-five minutes after filament introduction, mice breathed normobaric air, normobaric 100% O(2) (normobaric hyperoxia), or 100% O(2) at 3 ata (HBO) for 95 minutes. Hypoxic regions were mapped on tissue sections after preischemic infusion of the in vivo hypoxia marker EF-5. Hypoxia-inducible factor-1 alpha protein was measured after 2-hour middle cerebral artery occlusion using immunofluorescence and immunoblotting. Vascular endothelial growth factor expression was analyzed using in situ mRNA hybridization.


RESULTS
Severity of ischemia did not differ among groups. HBO (35.2+/-10.4 mm(2)) significantly reduced the area of EF-5-stained hypoxic regions in focal cerebral ischemia compared with normobaric hyperoxia (46.4+/-11.2 mm(2)) and air (49.1+/-8 mm(2), P<0.05, analysis of variance). Topographically, EF-5 fluorescence was decreased in medial striatum and in cortical ischemic border areas. Immunohistochemistry and immunoblotting revealed lower hypoxia-inducible factor-1 alpha protein in the ischemic hemisphere of HBO-treated mice. Moreover, mRNA in situ hybridization showed lower expression of vascular endothelial growth factor in HBO and normobaric hyperoxia groups.


CONCLUSIONS
Measurement of extrinsic and intrinsic markers of hypoxia revealed that HBO improves penumbral oxygenation in focal ischemia. Modification of the transcription factor hypoxia-inducible factor-1 alpha and its downstream targets may be involved in effects of HBO.
Sun
Li
L
Department of Neurology, Ruprecht-Karls-University Heidelberg, Heidelberg, Germany.
Marti
Hugo H
HH
Veltkamp
Roland
R
D000818:Animals; D001921:Brain / Q000378:metabolism; D002545:Brain Ischemia / Q000150:complications / Q000209:etiology / Q000378:metabolism* / Q000628:therapy*; D002540:Cerebral Cortex / Q000378:metabolism; D003342:Corpus Striatum / Q000378:metabolism; D015536:Down-Regulation; D017341:Etanidazole / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D005455:Fluorescent Antibody Technique; D006845:Hydrocarbons, Fluorinated / Q000493:pharmacokinetics; D006931:Hyperbaric Oxygenation; D002534:Hypoxia, Brain / Q000209:etiology* / Q000378:metabolism / Q000503:physiopathology*; D051793:Hypoxia-Inducible Factor 1 / Q000378:metabolism*; D007202:Indicators and Reagents / Q000493:pharmacokinetics; D020244:Infarction, Middle Cerebral Artery / Q000150:complications; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010100:Oxygen / Q000627:therapeutic use; D010101:Oxygen Consumption; D012333:RNA, Messenger / Q000378:metabolism; D013194:Staining and Labeling; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D005466:Fluorocarbons
D017341:Etanidazole; D006845:Hydrocarbons, Fluorinated; D051793:Hypoxia-Inducible Factor 1; D007202:Indicators and Reagents; D010100:Oxygen; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; C096011:2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl)acetamide; D005466:Fluorocarbons