| Objective:Alpha-synuclein(?-syn)is the characteristic pathological change for Parkinson's disease,and its abnormal aggregation is associated with the depletion of nigrostriatal dopaminergic neurons.Dysfunction of Wnt/?-catenin signaling pathway was reported to be associated with a variety of neurodegenerative diseases,including Alzheimer's disease,multiple sclerosis,and Huntington's disease.Data showed the activity of Wnt/?-catenin signaling pathway is decreased in PD and is associated with neurodegeneration.It was found that abnormal aggregation of ?-synuclein in PD can inhibit the transcriptional activation of NFAT and CREB,the key molecules of Wnt/?-catenin signaling pathway,suggesting that ?-synuclein can affect Wnt/?-catenin signaling pathway.This study was designed to investigate whether a-synuclein can interfere with the activity of neuronal cells in the pathogenesis of Parkinson's disease by inhibiting the Wnt/?-catenin signaling pathway in PD.Methods:1.Establishment of PD mouse model through MPTP injury,followed by the detection of the expression of ?-synuclein,T-GSK-3?,p-GSK-3? protein in the brain tissue.2.Flow cytometry was used to detect the expression of ?-synuclein and p-GSK-3? in SH-SY5Y cells in normal control group,MPP+injury group(1 mM MPP+treatment for 24 h)and ?-syn overexpression group(?-syn transfection).3.Western blotting was used to detect the expression of ?-synuclein,T-GSK-3? and p-GSK-3? protein in SH-SY5Y cells after aripiprazole was used as an agonist of Wnt/?-catenin signaling pathway for 24 h.4.Viability was analysed by MTT method to detect the protection effect of the Wnt/?-catenin signaling pathway agonist against ?-synuclein injury.5.Hoechest 33342/PI double staining method was used to detect different apoptosis level between groups.6.Laser scanning confocal microscope was used to detect the potential colocalization of ?-synuclein with GSK-3?.Results:1.There was no significant differences for the expression of ?-synuclein and p-GSK-3? between control grouped MPTP injured mice(P>0.05).There was no significant differences for the expression of T-GSK-3? between the three groups(P>0.05),The expression of a-synuclein in brain tissue of a-syn-transgenic mice was significantly higher than normal control and MPTP-injured group,and the difference was statistically significant(P<0.05).The level of p-GSK-3? protein in the brain tissue of a-syn transgenic mice was significantly higher than that of normal mice and MPTP-injured mice,and the difference was statistically significant(P<0.05).2.There was no significant differences for the expression of a-synuclein and p-GSK-3? between the normal control group and the MPP injury group.The expression of a-synuclein in the a-syn overexpressing SH-SY5Y cells was significantly higher than that in the normal control group and the MPP injury group.The difference was statistically significant(P<0.05).The p-GSK-3? protein in the a-syn overexpression group was significantly higher than the normal control group and the MPP injury group,and the difference was statistically significant(P<0.05).3.There were no significant differences in the expression levels of a-synuclein,T-GSK-3? and p-GSK-3? between the normal control group and the MPP injury group.Compared with the above two groups of cells,a-synuclein expression was significantly increased in the a-syn overexpression group(P<0.05),T-GSK-3? was not significantly different,and p-GSK-3? expression was increased,the difference was statistically significant.Significance(P<0.05).The expression of a-synuclein in aripiprazole treatment group was significantly higher than that in normal control group and MPP injury group(P<0.05),and there was no significant difference with a-syn overexpression group,while p-GSK-3? expression was higher than a-syn.The overexpression group decreased significantly,and the difference was statistically significant(P<0.05).4.Compared with the normal control cells,the cell activity of the MPP injury group was significantly decreased,and the cell activity of the a-syn overexpression group was also significantly decreased,while the cell activity of the aripiprazole pretreatment group was higher than that of the a-syn transgenic group.Statistical significance(P<0.05).5.Compared with normal control cells,the apoptosis of a-syn overexpression group was significantly increased,while the apoptosis of aripiprazole pretreatment group was lower than that of a-syn overexpression group,the difference was statistically significant(P<0.05).6.There is a colocalization relationship between the corresponding fluorescent signals of a-synuclein and p-GSK-3?Conclusion:The cellular damage caused by a-synuclein is associated with the Wnt/?-catenin signaling pathway,and a-synuclein participates in the pathogenesis of Parkinson's disease by inhibiting the Wnt/?-catenin signaling pathway. |
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