The role of hypoxia-inducible factor-1α, aquaporin-4, and matrix metalloproteinase-9 in blood-brain barrier disruption and brain edema after traumatic brain injury.
OBJECTIVE
The present study investigated the role of hypoxia-inducible factor-1α (HIF-1α), aquaporin-4 (AQP-4), and matrix metalloproteinase-9 (MMP-9) in blood-brain barrier (BBB) permeability alterations and brain edema formation in a rodent traumatic brain injury (TBI) model.


METHODS
The brains of adult male Sprague-Dawley rats (400-425 g) were injured using the Marmarou closed-head force impact model. Anti-AQP-4 antibody, minocycline (an inhibitor of MMP-9), or 2-methoxyestradiol (2ME2, an inhibitor of HIF-1α), was administered intravenously 30 minutes after injury. The rats were killed 24 hours after injury and their brains were examined for protein expression, BBB permeability, and brain edema. Expression of HIF-1α, AQP-4, and MMP-9 as well as expression of the vascular basal lamina protein (laminin) and tight junction proteins (zona occludens-1 and occludin) was determined by Western blotting. Blood-brain barrier disruption was assessed by FITC-dextran extravasation, and brain edema was measured by the brain water content.


RESULTS
Significant (p < 0.05) edema and BBB extravasations were observed following TBI induction. Compared with sham-operated controls, the injured animals were found to have significantly (p < 0.05) enhanced expression of HIF-1α, AQP-4, and MMP-9, in addition to reduced amounts (p < 0.05) of laminin and tight junction proteins. Edema was significantly (p < 0.01) decreased after inhibition of AQP-4, MMP-9, or HIF-1α. While BBB permeability was significantly (p < 0.01) ameliorated after inhibition of either HIF-1α or MMP-9, it was not affected following inhibition of AQP-4. Inhibition of MMP reversed the loss of laminin (p < 0.01). Finally, while inhibition of HIF-1α significantly (p < 0.05) suppressed the expression of AQP-4 and MMP-9, such inhibition significantly (p < 0.05) increased the expression of laminin and tight junction proteins.


CONCLUSIONS
The data support the notion that HIF-1α plays a role in brain edema formation and BBB disruption via a molecular pathway cascade involving AQP-4 and MMP-9. Pharmacological blockade of this pathway in patients with TBI may provide a novel therapeutic strategy.
Higashida
Tetsuhiro
T
Department of Neurological Surgery, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Kreipke
Christian W
CW
Rafols
José A
JA
Peng
Changya
C
Schafer
Steven
S
Schafer
Patrick
P
Ding
Jamie Y
JY
Dornbos
David
D
Li
Xiaohua
X
Guthikonda
Murali
M
Rossi
Noreen F
NF
Ding
Yuchuan
Y
D000077584:2-Methoxyestradiol; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology; D051401:Aquaporin 4 / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D001812:Blood-Brain Barrier / Q000378:metabolism / Q000503:physiopathology*; D001929:Brain Edema / Q000209:etiology / Q000378:metabolism / Q000503:physiopathology*; D001930:Brain Injuries / Q000150:complications / Q000378:metabolism / Q000503:physiopathology*; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors / Q000502:physiology*; D007797:Laminin / Q000378:metabolism; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000502:physiology*; D061965:Matrix Metalloproteinase Inhibitors; D008565:Membrane Proteins / Q000378:metabolism; D008911:Minocycline / Q000494:pharmacology; D023421:Models, Animal; D062793:Occludin; D010750:Phosphoproteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D062826:Zonula Occludens-1 Protein
D000888:Antibodies, Anti-Idiotypic; C496032:Aqp4 protein, rat; D051401:Aquaporin 4; C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D008565:Membrane Proteins; D062793:Occludin; C568987:Ocln protein, rat; D010750:Phosphoproteins; C569044:Tjp1 protein, rat; D062826:Zonula Occludens-1 Protein; D004958:Estradiol; D000077584:2-Methoxyestradiol; D020780:Matrix Metalloproteinase 9; D008911:Minocycline