The Study On Neuroprotective Mechanism Of Edaravone Enhances The Clearance Of α-synuclein Through Autophagy Activation In A Model Of Parkinson’s Disease

Objective:To explore the effects of edaravone on oxidative stress,autophagy and α-synuclein expression in Parkinson’s disease,as well as the neuroprotective mechanism of Eda clearing α-syn aggregation.Methods:The transgene Caenorhabditis elegans BZ555[Pdat-1::GFP],NL5901[Punc-54::α-synuclein::YFP+unc-119] and PC12 cells were induced by6-OHDA to establish the PD model,and the animals and cells were divided into control group,model group and Eda treatment group.(1)Animal experiments:Dopamine-dependent behavior and the morphology of DA neurons were observed to evaluate the DA system changes in PD animals.Oxidative stress levels of C.elegans in each group were detected and analyzed.The expression of α-syn protein in NL5901 strain in each group was detected,and the relative quantification was determined by Western Blot.(2)Cell experiment: MTT colorimetry was used to detect cell viability to determine the optimal modeling concentration of 6-OHDA and the optimal intervention concentration of Eda;The ROS level,MDA content and SOD activity of each group were detected to evaluate the oxidative stress level of cells.The expression levels of TH,α-syn and autophagy-related proteins(LC3,p62)in each group were detected by Western Blot and cellular immunofluorescence,as well as the expression levels of α-syn protein in cells treated with autophagy inhibitor 3-MA.Results:(1)Animal experiments: Compared with the control group,the dopaminedependent behavior of PD animals induced by 6-OHDA was significantly changed,which showed that the DA dependence was weakened.The morphological changes of dopaminergic neurons were characterized by shrinkage of cell body and discontinuous synapses.ROS level increased significantly.The α-syn protein was dispersed and its expression was increased.Compared with the model group,the dopamine-dependent behavior of C.elegans was recovered after Eda treatment,and the neurons morphology was close to normal.ROS level decreased.The expression ofα-syn protein was decreased.(2)Cell experiment: Compared with the control group,ROS level,MDA content and SOD activity in model group were increased,which reflected the increase of oxidative stress level.The decrease of TH protein expression in cells reflects the decrease of its secretion function.α-syn expression increased,LC3 expression decreased,and p62 expression increased.Compared with model group,after Eda treatment,ROS level decreased,MDA content decreased,SOD activity increased,reflecting the reduction of oxidative stress level;The expression of TH protein increased,which reflected the recovery of secretion function.α-syn expression decreased,LC3 expression increased,p62 expression decreased.After the addition of autophagy inhibitor 3-MA on the basis of Eda,the expression of α-syn was increased,which reflected that 3-MA blocked the degradation of α-syn aggregation by autophagy.Conclusions:(1)Eda can reduce the level of oxidative stress in PD model,reduce the aggregation of α-syn protein,maintain the morphology and function of dopaminergic neurons,and have a neuroprotective effect on PD.(2)Eda can activate autophagy and promote autophagy clearance of α-syn protein aggregation.