The Study On The Role Of FOXO3a In Autophagy And Apoptosis In The Parkinson's Model Of SH-SY5Y Induced By Rotenone

BackgroundParkinson's disease(PD)is a global chronic neurodegenerative disease with various etiologies and pathologic reasons for apoptosis of brain dopaminergic neurons.Lewy body,as the typical pathological manifestations of the disease in the residual cells,was concerned by researchers.Lewy body is the product of the abnormal degradation of the a-synuclein after the abnormal protein metabolism in the neurons..The molecular mechanism of autophagy is related to various mechanisms such as metabolism,immunity,nutrition,stress and death of cells.In a variety of diseases,including Parkinson's disease,autophagy in the cellular level of molecular regulation mechanism and its effects on the disease is very extensive.The lewy body is actually a special pathological form of autophagic lysosomes in cells..In associated with further study,because of the particularity of dopaminergic neurons in the disease,animal experiments of inducer in the corresponding cell tests are difficult to replicate a satisfactory autophagy cell modelMethod and ResultsThrough literature retrieval,the researchers first induced different concentration of rotenone on SH SY5Y cells in the different periods.After fluorescence staining microscopic quantitative apoptotic cells observation,the count of cell activity test verification method(CCK8),autophagy protein determination after induction,fluorescence staining observation,lysosomal inhibitors reverse authentication and so on a variety of experimental methods,.Eventually we selected specific concentrations of rotenone in a specific time on SH SY5Y cells for different researches focus on the center of the molecular mechanism of autophagy.purpose of early Parkinson's disease.In the further study of the cell model,we found that the FOXO3a protein,as a kind of complex function of transcription factors,play an important role.through the movement in cell by phosphorylation both in the autophagy and apoptosis of cells.After transfection of the overexpression plasmid and FOXO3a-siRNA interference of the cell model,we tested the apoptosis of the PD cell model after qPCR and protein detection.Through the above experiments,we believe that the expression of FOXO3a protein in the cell model plays an important role in promoting apoptosis.After that,we used the PD cell model to conduct further screening test analysis on the possible effected molecular of FOXO3a upstream.The results suggest that the phosphorylation of JNK may play an important role in the apoptosis pathway of the transcription factor FOXO3a.Finally,we used p-JNK inhibitor and siRNA-FOXO3a to determine the expression of FOXO3a protein and the measurement of cell death in the PD cell model,and finally the following main conclusions were obtained:1.At a particular time,concentration of rotenone on the SH SY5Y cells can be used as a satisfactory cell model,to simulate the early Parkinson's disease nerve cells in the pathological changes,and then start the further study on the molecular mechanism of autophagy;2.Flow cytometry and CCK8 methods were used to confirm that the latter could be used to detect the cell activity of the Parkinson's cell model to accurately reflect the pathological state of the cell.3.The hyperexpression of FOXO3a in the PD cell model promotes apoptosis,and the phosphorylation activation of JNK plays a key role.4.The inhibitor of p-JNK and FOXO3 protein interfering RNA can decrease the apoptosis of the cell model and may provide a new method for the treatment of PD.The above conclusions are not reported in the relevant cell model studies.