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Objective:To study the expressione of autophagy-related protein Beclin-1 and LC3 in cortex and hippocampus of rats after cerebral ischemia reperfusion injury and the efficacy of Edaravone.Methods:Sprague Dawley rats were randomly divided into sham group,model group and Edaravone group.The cerebral ischemia reperfusion model was induced via Zea Longa with blocking the middle cerebral artery of 2 h and reperfusing of 24 h.Animals assigned to sham group were only separated left common carotid artery.Edaravone was injected intraperitoneally at a dose of 1 0 mg/kg at 15 min before reperfusion.The condition of nerve injury of rats was conducted by Neurobehavioral score.The degree of brain injury and success of model were determined based on 2,3,5-triphenyltetrazolium chloride(TTC)staining.The changes of neuron stained by hematoxylin and eosin (HE) in cortex and hippocampus were observed.The expression of Beclin-1 and LC3 was measured by immunohistochemistry.Results:After cerebral ischemia reperfusion the neurobehavioral score of edaravone group was(2.00± 0.67),which was obcviously less than(2.50± 0.53) of model group(P<0.05).The infraction focus and the neuron injury in cortex and hippocampus neurons were also observed in model group,and the edaravone group reduced above expression.The positive rate of Beclin-1 of each group in cortex were (1 1.08± 0.85)%,(33.42± 1.57)% and (25.61± 1.39)%,there was significant difference between model group with sham group and edaravone group (P<0.05).The positive rate of Beclin-1 of each group in hippocampus were (10.34± 0.21)%,(31.82± 1.73)% and (22.74± 1.26)%,there was significant difference between model group with sham group and edaravone group(P < 0.05).The positive rate of LC3 of each group in cortex were (15.33± 0.47)%,(39.72± 1.73)% and (28.53± 1.61)%,there was significant difference between model group with sham group and edaravone group(P<0.05).The positive rate of LC3 of each group in hippocampus were (13.74± 0.37)%,(32.53± 1.43)% and(25.38± 1.23)%,there was significant difference between model group with sham group and edaravone group (P<0.05).Conclusion:The expression of Beclin-1 and LC3 was increased after cerebral ischemia reperfusion.Edaravone may reduce autophagy and brain injury through downregulation the expression of Beclin-1 and LC3.
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[ABSTRACT]AIM:Toinvestigatetheroleofcysteine-rich61(Cyr61/CNN1)inproliferationandmigrationof bone marrow mesenchymal stem cells ( BMSCs ) .METHODS: The lentiviral vector carrying CCN 1 ( Lenti-GFP-CCN1 ) was constructed and then transfected into the rat BMSCs .The cells were divided into non-transfection group , transfection group ( transfected with Lenti-GFP-CCN1 ) and negative control group ( Lenti-GFP ) .The fluorescence intensity of the transfected BMSCs was observed under inverted fluorescence microscope .The effects of CCN1 on the proliferation and mi-gration of BMSCs were detected by MTT assay and scratch wound healing assay .RESULTS:The proliferation of BMSCs transfected with Lenti-GFP CCN1 had no significant difference compared with negative control group and control group .The width/thickness ratio of migrated BMSCs in wound healing was significantly higher in Lenti-GFP-CCN1 group than that in negative control group and control group (P<0.05).CONCLUSION:Exogenous CCN1 promotes the migration of BMSCs.