MCAO model and vinpocetine treatment

The middle cerebral artery occlusion (MCAO) model was employed as described in our previous study.[14] Briefly, the mice were first anesthetized with 5.0% isoflurane. Then, an incision was made in the skin and the right common carotid artery (CCA), external carotid artery (ECA), and internal carotid artery (ICA) were carefully exposed. Microvascular aneurysm clips were applied to the right CCA and the ICA. A coated 6-0 filament (Doccol, Redlands, CA) was introduced into an arteriotomy hole, fed distally into the ICA, and advanced a predetermined distance located 8 mm from the carotid bifurcation toward the MCA. After a 60-min period of focal cerebral ischemia, the filament was gently removed (onset of reperfusion). The collar suture at the base of the ECA stump was then tightened. The skin incision was closed and anesthesia was discontinued. Sham mice underwent neck dissection and coagulation of the ECA, but the MCA was not occluded. The success rate of the MCAO operation was approximately 90%. The animals were randomly divided into groups according to computer-generated randomization schedules, and blinding to group allocation was ensured. Dead animals and animals in which the MCAO model failed were excluded from the experiments. The animals were intraperitoneally injected with vinpocetine (10 mg/kg) after MCAO.[21, 25] The 10 mg/kg dose was selected for both compounds as optimal dose based on previous in vivo studies.