Middle Cerebral Artery Occlusion and Reperfusion

Left middle cerebral artery occlusion followed by reperfusion was conducted as we previously reported [29 (link)]. In brief, the mice were anesthetized with 2.5% isoflurane. A midline neck incision was used to expose the left external carotid artery (ECA), internal carotid artery (ICA), and common carotid artery (CCA). The ECA is ligated using two surgical 7–0 threads. One was toward the distal end of the ECA, while the other was near the proximal end, as close to the bifurcation of the ECA and ICA as feasible, followed by an incision between the ECA ligatures. Finally, a 2-cm long nylon filament with a diameter of 0.25 ± 0.03 mm was introduced into the ICA through the external carotid stump and progressed to occlude the origin of the middle cerebral artery (MCA). Cerebral blood flow (CBF) was monitored using a Laser Speckle Imaging System (RFLSI III, RWD Life Science, Shenzhen, China). The nylon filament was gently extracted from the ECA after an hour of ischemia for reperfusion. The incision at the neck was then closed. All of the animals were resuscitated on heat pads for two hours before being kept separately for additional investigation. The sham group mice underwent the same surgical operation technique as the control mice but without vaso-occlusive.