GSK-3? To Adjust The Role Of Autophagy In Parkinson's Disease Nerve Cells And Mechanism Research

Objective : Screening of stable express A30 P a-synuclein PC12 cell as the research object, through the experiment of cellular and molecular level, reliminary discussion to express the activity of different GSK3? whether for the regulation of autophagy pathway causes a-synuclein autophagy level change and its effects on cell growth.Methods:1.Use the vector of A30 P a-synuclein transfect to good condition of PC12 cells, and by G418 screening stable expression cell.2. Liposome method will GSK3? expression plasmid, GSK3?silence plasmid and blank plasmid transfection these cell lines, screening stable express different the activity of GSK3? cell lines, Werstern blot detecting the expression of GSK3? after transfection and verify the transfection efficiency.3. Western blot test groups of cells in LC-3?, Beclin1, a-synuclein expression.4. Determined by MTT method to detect the change of cell proliferation.5. TUNEL method to detect apoptosis.Results : 1.GSK3? express group compared with GSK3? silence group,blank plasmidgroup, and blank control group: the expression of LC-3? and Beclin1 are increases(P<0.05);the expression of a-synuclein is reduced(P<0.05);the results of MTT and TUNEL show that the cells is apoptosis decrease and survival increase(P<0.05).2.GSK3? silence group compared with GSK3? express group,blank plasmidgroup, and blank control group: the expression of LC-3? and Beclin1 are reduce(P< 0.05); the expression of a-synuclein is increased(P<0.05);the results of MTT and TUNEL show that the cells is apoptosis increase and survival reduce(P<0.05).Conclusion: GSK3? can increase the level of autophagy of stable express A30 P a-synuclein PC12 cell, which would be helpful to the survival of cells.