The Effect Of Butylphthalide Through P53/mTOR Pathway On Autophagy After Cerebral Ischemia-reperfusion Injury In Rats

Objective:To explore whether butylphthalide can affect the autophagy reaction after cerebral ischemia-reperfusion in rats through p53/mTOR pathway,and play a neuroprotective role in improving the symptoms of cerebral infarction.Methods:36 adult male SD rats were randomly divided into sham operation group,butylphthalide group,model group and 12 rats in each group.In the model group and the treatment group,the rat middle cerebral artery occlusion and reperfusion(MCAO)model was made by the thread thrombus method.The sham operation group only separated the blood vessels without inserting the thread thrombus.The treatment group was given butylphthalide solution(0.1g Butylphthalide Soft Capsules dissolved in 100 ml Tween80 solution)intraperitoneally(80mg/kg)after ischemia 1H,and the model group was injected with equal volume Tween80 solution after ischemia 1H.Reperfusion after 2 h of cerebral ischemia and death after 24 hours.The neurological function was evaluated by Longa score method.TTC staining was used to detect infarct volume of rats,using immunohistochemical method(SABC method)to detect brain tissue of rats in each group were p53,mTOR,Beclin-1,Lc3 expression level.And we should observe the morphology of brain tissue cells modified by HE staining,electron microscope observation of autophagosome formation.The experimental data were compared with the average number of standard deviation,the multiple groups were compared with the variance analysis,the two samples were compared with the t test,and the difference of P < 0.05 was statistically significant.Results:In the sham operation group,there were no symptoms such as limb movement,walking deviation,and other nervous system defects.The neurological deficit score in the treatment group was lower than that in the model group.There was no infarction lesion in the sham operation group.Cerebral infarction volume in the model group was larger than in the treatment group.Compared with that of the sham operation group,p53,mTOR,Beclin-1 and LC3 positive cells in the model group and treatment group were increased(P < 0.05).The treatment group p53 positive cells expression significantly lower than model group(P<0.05).The treatment group the number of mTOR positive cells was significantly higher than the model group.Beclin-1?LC3 positive cells in the treatment group was significantly lower than that of model group(P < 0.05).HE staining of nerve tissue of rats in the sham operated group with clear structure,clear structure of neurons,cell membrane integrity,homogeneous cytoplasmic staining,nuclei clear and complete;model group nerve tissue structure visible damage,osteoporosis,damage and reduce the number of neurons,structure,visible swelling of neurons,nuclear condensation,pyknosis,dissolution,nucleolus the nuclear membrane disappeared,cytoplasmic stain;treatment group visible tissue damage,osteoporosis,and reduce the number of neurons or swelling,nucleus condensation,pyknosis,nuclear dissolution,nucleolus disappeared,cytoplasm becomes shallow,but the extent was significantly reduced compared with model group.Conclusion:Butylphthalide has a protective effect on cerebral ischemia reperfusion injury and may play a role in reducing autophagy through the P53/mTOR pathway.