Na(+)-K (+)-2Cl (-) cotransport inhibitor attenuates cerebral edema following experimental stroke via the perivascular pool of aquaporin-4.
BACKGROUND
The Na(+)-K(+)-2Cl(-) cotransporter localized in the brain vascular endothelium has been shown to be important in the evolution of cerebral edema following experimental stroke. Previous in vivo studies have demonstrated that bumetanide, a selective Na(+)-K(+)-2Cl(-) cotransport inhibitor, attenuates ischemia-evoked cerebral edema. Recently, bumetanide has been shown to also inhibit water permeability via aquaporin-4 (AQP4) expressed in Xenopus laevis oocytes. We tested the hypothesis that the perivascular pool of AQP4 plays a significant role in the anti-edema effect of bumetanide by utilizing wild-type (WT) mice as well as mice with targeted disruption of alpha-syntrophin (alpha-Syn(-/-)) that lack the perivascular pool of AQP4.


METHODS
Isoflurane-anesthetized adult male WT C57Bl6 and alpha-Syn(-/-) mice were subjected to 90 min middle cerebral artery occlusion (MCAO) followed by 24 or 48 h of reperfusion. Adequacy of MCAO and reperfusion was monitored with laser-Doppler flowmetry over the ipsilateral parietal cortex. Infarct volume (tetrazolium staining), cerebral edema (wet-to-dry ratios), and AQP4 protein expression (immunoblotting) were determined in different treatment groups in separate sets of experiments.


RESULTS
Bumetanide significantly attenuated infarct volume and decreased ipsilateral hemispheric water content in WT mice compared to vehicle treatment. In alpha-Syn(-/-) mice, bumetanide treatment had no effect on infarct volume or ischemia-evoked cerebral edema. Bumetanide-treated WT mice had a significant attenuation of AQP4 protein expression at 48 h post-MCAO compared to vehicle-treated WT mice.


CONCLUSIONS
These data suggest that bumetanide exerts its neuroprotective and anti-edema effects partly via blockade of the perivascular pool of AQP4 and may have therapeutic potential for ischemic stroke in the clinical setting.
Migliati
Elton R
ER
Department of Neurology, Oregon Health & Science University, Portland, OR, USA.
Amiry-Moghaddam
Mahmood
M
Froehner
Stanley C
SC
Adams
Marvin E
ME
Ottersen
Ole Petter
OP
Bhardwaj
Anish
A
D000818:Animals; D051401:Aquaporin 4 / Q000037:antagonists & inhibitors*; D001929:Brain Edema / Q000209:etiology* / Q000503:physiopathology*; D002034:Bumetanide / Q000494:pharmacology*; D002544:Cerebral Infarction / Q000150:complications / Q000209:etiology / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018696:Neuroprotective Agents / Q000494:pharmacology*; D049994:Sodium Potassium Chloride Symporter Inhibitors / Q000494:pharmacology*; D020521:Stroke / Q000150:complications* / Q000209:etiology; D051844:alpha-Synuclein / Q000172:deficiency
D051401:Aquaporin 4; D018696:Neuroprotective Agents; D049994:Sodium Potassium Chloride Symporter Inhibitors; D051844:alpha-Synuclein; D002034:Bumetanide
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