Rat ischemia stroke model

The rat ischemia stroke model was established according to a previous report [11]. Adult male Wistar rats (weight: 246 ± 10 g) were purchased from the Chinese Academy of Sciences (Shanghai, China). Rats were bred and maintained in a pathogen-free facility and were subjected to middle cerebral artery occlusion (MCAO) as previously described [11]. After anesthesia by sodium pentobarbital (100 mg/kg), the right common carotid artery, external carotid artery, and internal carotid artery of rats were exposed through a midline cervical incision. A piece of monofilament nylon suture was inserted through the right internal carotid artery to the base of the middle cerebral artery, resulting in preventing blood flow to the cortex and striatum for 90 min. The rats were then recovered for 1 day. The total brains were then removed for downstream assays of the present study. The sham control rats were subjected to similar surgeries to expose the carotid arteries but without occlusion of the middle cerebral artery. The present study was approved by the Animal Care Committee at the Third Central Hospital of Tianjin. Animals were operated in accordance with the institutional guidelines for animal care.