Establishing middle cerebral artery occlusion and reperfusion (MCAO/R) model

Male Sprague-Dawley rats weighing approximately 320–350 g (10–12 weeks) (n=16 per group) were obtained from Shanghai Lab Animal Research Center, Shanghai, China. The rats were under a 12 hours light cycle with food and drinking water available ad libitum. The experimental procedures were approved by Shandong Provincial Hospital Affiliated to Shandong University Animal Ethics Committee (2003-045). Transient middle cerebral artery occlusion (MCAO) using an intraluminal suture was induced under isoflurane anesthesia as previously described [14]. Reperfusion was accomplished by withdrawing the suture after 60 min of ischemia. The brain reperfusion was allowed for 0, 0.5, 2, 4, 6, 8, 12 and 24. Sham-operated rats underwent identical surgery except that the monofilament nylon suture was not inserted. After incision closure, the rats were placed into cages to recover, with free access to food and water. Rat rectal temperature was monitored and maintained at 37.0±0.5°C during the whole procedure.