Middle cerebral artery occlusion (MCAO) was used to induce cerebral I/R injury, as described previously. 20 Following anaesthetization by pentobarbital sodium (40 mg/kg) and buprenorphine‐HCl (0.1mg/kg) intraperitoneally, rats were kept on a thermostatic operating table at 37°C. After sterilizing the towel, the right neck skin was cut open to fully display the common carotid artery (CCA), internal carotid artery (ICA) and external carotid artery (ECA). Then, A 4‐0 line embolism (diameter with coating 0.37 ± 0.02mm; Doccol Corporation, Sharon, MA, USA) was sliped into the CCA and advanced ICA by 18.5 ± 0.5mm distance. After 90 min of ischaemia, the line embolism was removed for blood reperfusion. Rats assessed by Longa‐Z method without neurologic impairment after reperfusion (neurological function score < 1 or > 3) were excluded from this study.

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