Protecting against cerebrovascular injury: contributions of 12/15-lipoxygenase to edema formation after transient focal ischemia.
OBJECTIVE
The concept of the neurovascular unit suggests that effects on brain vasculature must be considered if neuroprotection is to be achieved in stroke. We previously reported that 12/15-lipoxygenase (12/15-LOX) is upregulated in the peri-infarct area after middle cerebral artery occlusion in mice, and 12/15-LOX contributes to brain damage after ischemia-reperfusion. The current study was designed to investigate 12/15-LOX involvement in vascular injury in the ischemic brain.


METHODS
In cell culture, a human brain microvascular endothelial cell line was subjected to either hypoxia or H(2)O(2)-induced oxidative stress with or without lipoxygenase inhibitors. For in vivo studies, mice were subjected to 90 minutes middle cerebral artery occlusion, and the effects of either 12/15-LOX gene knockout or treatment with lipoxygenase inhibitors were compared. Expression of 12/15-LOX and claudin-5 as well as extravasation of immunoglobulin G were detected by immunohistochemistry. Edema was measured as water content of brain hemispheres according to the wet-dry weight method.


RESULTS
Brain endothelial cells were protected against hypoxia and H(2)O(2) by the lipoxygenase inhibitor baicalein. After focal ischemia, 12/15-LOX was increased in neurons and endothelial cells. The vascular tight junction protein claudin-5 underwent extensive degradation in the peri-infarct area, which was partially prevented by the lipoxygenase inhibitor baicalein. Leakage of immunoglobulin G into the brain parenchyma was significantly reduced in 12/15-LOX knockout mice as well as wild-type mice treated with baicalein. Likewise, brain edema was significantly ameliorated.


CONCLUSIONS
12/15-LOX may contribute to ischemic brain damage not just by causing neuronal cell death, but also by detrimental effects on the brain microvasculature. 12/15-LOX inhibitors may thus be effective as both neuroprotectants and vasculoprotectants.
Jin
Guang
G
Massachusetts General Hospital, Neuroprotection Research Laboratory, 149 13th Street, R. 2401, Charlestown, MA 02129, USA.
Arai
Ken
K
Murata
Yoshihiro
Y
Wang
Sophia
S
Stins
Monique F
MF
Lo
Eng H
EH
van Leyen
Klaus
K
D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D001093:Arachidonate 15-Lipoxygenase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism; D001929:Brain Edema / Q000201:enzymology* / Q000503:physiopathology / Q000517:prevention & control; D020520:Brain Infarction / Q000188:drug therapy / Q000201:enzymology* / Q000503:physiopathology; D002545:Brain Ischemia / Q000188:drug therapy / Q000201:enzymology* / Q000503:physiopathology; D002478:Cells, Cultured; D062507:Claudin-5; D019610:Cytoprotection / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use; D044950:Flavanones / Q000627:therapeutic use*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007074:Immunoglobulin G / Q000378:metabolism; D002546:Ischemic Attack, Transient / Q000188:drug therapy / Q000201:enzymology* / Q000503:physiopathology; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism
C475064:12-15-lipoxygenase; C107466:CLDN5 protein, human; D062507:Claudin-5; D004791:Enzyme Inhibitors; D044950:Flavanones; D007074:Immunoglobulin G; D008565:Membrane Proteins; C006680:baicalein; D006861:Hydrogen Peroxide; D001092:Arachidonate 12-Lipoxygenase; D001093:Arachidonate 15-Lipoxygenase