The Effects Of Human Urinary Kallidinogenase On Focal Cerebral Ischemical Reperfusion Injury In Rats

Part I Effect of human urinary kallidinogenase on neuronal apopto-sis at focal cerebra injured region after ischemia-reperfusion in ratsObjective:To investigate the effect of human urinary kallidinogenase (HUK) on the expression of caspase-3, apoptosis-inducing factor (AIF), Survivin and neuronal apoptosis in perifocal areas of cerebra after ischemia-reperfusion (I/R) in rats.Methods:Wistar rats were randomly divided into sham group,model group and HUK group.The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by thread ligation. After operation in sham group,2 h ischemia and 6h,24h, 48h,72h reperfusion in model and HUK group,the expression of caspase-3,AIF, Survivin positive cells and neuronal apoptosis in the ischemic cortex were detected by immunohistochemistry and in situ apoptotic detection (TUNEL method).Results:The caspase-3,AIF,Survivin and Tunel positive neurons expresses in perifocal areas at different time points after I/R. The trends of expressed caspase-3, AIF,Survivin and Tunel positive neurons in HUK group and model group were basi-coincidence. The peak of expressed caspase-3,AIF and Tunel positive neurons were at 24h in perifocal areas after I/R.The peak of expressed Survivin was at 48h in perifocal areas. The optical density value of Caspase-3,AIF and Tunel positive neurons in HUK group at each time were significantly decreased compared with model groups,and the optical density value of Survivin was increased compared with model groups (P<0.05).Conclusions:HUK play protective function for the cerebral tissues after I/R by increasing the expression of Survivin, reducing the expression of caspase-3,AIF and tardive neuronal apoptosis in posse. Part II Effect of human urinary kallidinogenase on function of spatial learning and memory in rats and nestin expression in peri-infarction cortex after focal cerebral ischemia-reperfusionObjective:To investigate the effects of human urinary kallidinogenase (HUK) on the ability of spatial learning and memory in rats and the expression of nestin in peri-infarction cortex after focal cerebral ischemia-reperfusion.Methods:The focal cerebral Ischemia-reperfusion model was established by introducing an intraluminal filament into the right middle cerebral artery of the rats, then HUK was administered introperitoneally after the operation and afterward once a day for 2 weeks. The spatial learning and memory function was studied by morris water maze test,Then Nissl staining technique was conducted for observing the morphology of neurons in peri-infarction cortex,the nestin expression in the peri-infarction cortex was measured by immunohistochemistry.Results:The ischemia-control group exhibited seriously spatial learning and memory deficits in both place navigation trail and spatial probe trial.In place navigation trial, the mean values of escape latency of middle-dose and higher-dose HUK groups were shorter than those of ischemia-control group (P<0.05).In spatial probe trial, there were significant difference in the time percentage of spending in the former platform quadrant and frequency of crossing the former platform site between middle and higher-dose HUK groups and ischemia-control group(P<0.05). Nissl staining analysis showed that the number of neurons in the peri-infarction cortex of middle and higher-dose HUK groups significantly increased as compared with that of ischemia-control group (P<0.05).Immunohistochemical analysis showed that nestin expression in the peri-infarction cortex of middle and higher-dose HUK groups significantly increased as compared with that of ischemia-control group (P<0.05).Conclusion:HUK improves the rats spatial learning and memory ability after focal cerebral ischemia-reperfusion, this may be the results of the increasing expression of nestin and the proliferation of neural stem cells after focal cerebral ischemia.