GSK3β to adjust the role of autophagy in Parkinson's disease nerve cells and mechanism research / 中国医师杂志

ABSTRACT

Objective To screen stable express A30P a-synuclein PC12 cell as the research object,and to preliminarily investigate the activity of different glycogen systhesis kinase 3β (GSK3β) for the regulation of autophagy pathway causes a-synuclein autophagy level change and its effects on cell growth.Methods The vector of A30P a-synuclein was used to transfect to PC12 cells.G418 was used to screen stable expressed cells.Liposome method was used to transfect GSK3β-expressed plasmid,GSK3β silence plasmid,and blank plasmid into these cell lines,and stable expressed activity of GSK3β cell lines were screened.Western blot was used to detect the expression of GSK3 β after transfection and verify the transfection efficiency.Western blot was used to test groups of cells in LC-3 Ⅱ,Beclin1,and a-synuclein expressions.Methyl thiazolyl tetrazolium (MTF) method was used to detect the change of cell proliferation.Terminal dexynucleotidyl transferase (TdT) mediated dUTP nick end labeling (TUNEL) method was used to detect apoptosis.Results (1) When GSK3β expressed group was compared to GSK3β silence group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly increased (P ＜ 0.05),the expression of a-synuclein was significantly reduced (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the decrease of apoptosis and increase of cell proliferation,with a statistically significant difference (P ＜ 0.05).(2) When GSK3β silence group was compared to GSK3β express group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly reduced (P ＜ 0.05),the expression of a-synuclein was significantly increased (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the increase of apoptosis and decrease of cell proliferation,with a statistically significant difference (P ＜0.05).Conclusions (1) Activation of GSK3β activity can improve the level of cell autophagy,enhance the ability of alpha-synuclein degradation,and promote cell survival.(2) Inhibition of GSK3β activity can reduce the level of cell autophagy,weaken the ability of alpha-synucleindegradation,and reduce cell survival.(3) Autophagy is closely related to the pathogenesis of Parkinson's disease (PD).The improvement of the level of autophagy and enhancing of the degradation of asynuclein is a new way of the future treatment of PD.