The Effects Of NR2B,Endo G On Focal Cerebral Ischemical Reperfusion Injury In Rats

PartⅠThe expression of NR2B,endoG after focal cerebral ischemia in ratsObjective:To investigate the regular expression of the N-methyl-D-aspartate receptor subunit NR2B protein and cell apoptosis-inducing factor DNA endonuclease G(endonuclease G,endo G)in basement Festival of brain in rat after focal cerebral ischemia-reperfusion,and infarct volume and nerve cell apoptosis.To explore the injury mechanism of nerve cells after cerebral ischemia-reperfusion.Methods:Totally 63 Weight 250 to 300 grams healthy adult male Wistar rats were randomly divided into:normal group(A),sham-operated group(B), cerebral ischemia-reperfusion(C).The middle cerebral artery occlusion (MCAO)rat models were made using intraluminal filament method.Using immunohistochemistry and image analysis method observed the expression of NR2B protein and endo G at 1h of focal cerebral ischemia and 1,6,24,72h after reperfusion in the basal ganglia.Detecting the infarct volume by TTC method,and the nerve cell apoptosis by TUNEL method.Results:cerebral ischemia-reperfusion can cause ischemic neuronal cell apoptosis or necrosis,the infarction volume was significantly increased than normal group and sham-operated group,the number of apoptotic cells reached the peak at cerebral ischemia-reperfusion 24h,and the apoptotic cells were seen around the necrosis area,this was almost unanimously with the positive expression of endoG;At ischemia-reperfusion 1h,the expression of NR2B receptor was decreased.With cerebral ischemia-reperfusion time was lasting,the positive expression of NR2B also gradually increased,it reached the peak at reperfusion 6h,and then declined sharply,continuing to 72h and after that.After ischemia-reperfusion 1h,the expression of endoG protein begun to increase,with cerebral ischemia-reperfusion time was lasting,the positive expression of NR2B gradually increased,reached the peak at reperfusion 24 h,and then decline gradually,continuing to 72h.Conclusion:The expression of NR2B was decreased in the early stage of reperfusion,at following stage it suffered change of increase and up to recovery;The expression of endoG is increased in the early stage of reperfusion,and then rapidly decreased.The expression both displayed time-dependent.Cerebral ischemia-reperfusion can cause ischemic neuronal cell apoptosis or necrosis.PartⅡThe effect of ifenprodil tartrate,ginsenoside-Rd and allied application on cerebral ischemia-reperfusion injury in ratsObjective:To investigate the neural protection of ifenprodil tartrate ginsenoside-Rd and allied application.Methods 117 healthy male adult wistar rats were randomly divided into 5 groups,the normal group,the model group,the ifenprodil tartrate-treated group,the ginsenoside-Rd-treated group and the allied application group.The ifenprodil tartrate-treated group were intraperitoneal injection with 2mg/kg of ifenprodil tartrate by preparing appropriate concentration solution;The ginsenoside-Rd-treated group were intraperitoneal injection with 2mg/kg of ifenprodil tartrate by preparing appropriate concentration solution temporaryly; The allied application group were intraperitoneal injection with the above both drugs at the same time.One time / a day,lasting for 3days before the model of middle cerebral artery occlusion(MCAO)was established.Then the rats were decapitated at 1hrs,6hrs,24hrs and 72hrs after reperfusion.The neurological deficit score was graded by Garcia's methods,the volume of the cerebral infarction was estimated by 2%TTC staining,HE staining, the number of nerve cells apoptosis by TUNELL,and the expression of NR2B endoG were investigated by immunohistochemistry.Results:1)The neurological deficit scores,the significant difference existed on the scores between the model group and the ginsenoside-Rd-treated group at 24hrs(P＜0.05),and the allied application group at 24hrs(P＜0.01).The difference between the drug-treated groups and the model group was not significant(P＞0.05)at other time points.2)The drug group obviously cut down cerebral infarct sizes(P＜0.01).3)Compared with model group, the drug-treated group can significantly reduce the positive expression of apoptosis neural cell at 6hrs,24hrs,72hrs(P＜0.01).the drug-treated groups compared with eath other,The allied application group can significantly reduce the number of nerve cell apoptosis(P＜0.01).4)HE staining,the drug-treated groups compared with the model group,the nerve cells of the model group decreased significantly,the remnants of nerve cell soma were narrowed and deformed,pyknosis,deeply stained,nucleolus, heterochromatin disappear.The interstitial edema were obvious.But the neuronal structure of the drug-treated were more integrity,Its morphology were relatively normal,interstitial edema were lighter.5) The expression of NR2B,compared with the model group,The simple drug group and the allied application group both can significantly reduce the expression of NR2B protein in basal ganglia area in rats at 6hrs(P＜0.01), but its reduction in the allied application group was more obvious.6) The expression of endoG,compared with the model group,the simple drug group and the allied application group both may reduce the expression of endoG,the difference is statistically significant at 6hrs,24hrs,72hrs(P＜0.05 or P＜0.01),while the role of the allied application group was more obvious. Conclusion:1.Ifenprodiltartrate and the ginsenoside-Rd can reduce neuronal apoptosis, and narrow cerebral infarct sizes and improve the neurological deficit scores,so it have a neuroprotective effect on impaired neuron in cerebral ischemia-reperfusion injury.2.Ifenprodiltartrate and the ginsenoside-Rd can reduce the expression of NR28 and endo G,block nerve cells necrosis and apoptosis from different segments.3.Allied application of Ifenprodiltartrate and the ginsenoside-Rd may be have a joint action,may diminish the the volume of cerebral infarct sizes,play an important neuroprotective effect on focal cerebral ischemia-reperfusion injury.