2.4. Preparation of the Middle Cerebral Artery Occlusion (MCAO) Model

The MCAO rat model was established according to previous studies [9, 10]. Briefly, rats were deeply anesthetized with 3% pentobarbital sodium (45 mg/kg) by intraperitoneal injection. A midline incision was made for blunt separation of the right common carotid artery (CCA), external carotid artery (ECA), and internal carotid artery (ICA). A 40 mm nylon thread plug (RWD Life Science Co., Ltd., Shenzhen, China) was inserted approximately 18 ± 2 mm into the ICA from the CCA. The proximal and distal ends of the CCA were ligated to prevent bleeding, and the wound was sutured. The rats were placed at a constant temperature of 37°C until they regained consciousness.