Rat Model of Focal Cerebral Ischemia/Reperfusion

Animal protocols were approved by the Institutional Animal Care and Use Committee of Xuanwu Hospital of Capital Medical University and in accordance with the principles outlined in the NIH Guide for the Care and Use of Laboratory Animals. Male Sprague-Dawley rats (280–300g) were anesthetized with 3.5% enflurane in N₂O:O₂ (70%:30%). Focal cerebral ischemia was induced using the modified intraluminal filament method. Female rats were not used since the current study was designed to provide the proof of concept for the interaction of zinc with ROS. The animals underwent 90min of right middle cerebral artery occlusion (MCAO) and then were reperfused for 6 or 24h by withdrawal of the filament, as we previously described.⁴ Success of MCAO model was assessed by observing animal circling to the non-ischemic side at the end of ischemia, and further confirmed by 2,3,5-triphenyltetrazolium chloride (TTC) staining at the end of reperfusion.