Effects Of Donepezil On Amyloid Precursor Protein/alpha-Ampyloid Processing In Pheochromocytoma Cells

ObjectiveTo observe the effect of donepezil on amyloid precursor protein(APP) non-amyloid metabolic pathways in PC12cells and explore thepossible cell signaling mechanisms.MethodsThe PC12cells were divided into5groups: The control group,donepezil20μmol/L group, donepezil20μmol/L and PI3-Kinhibitor LY29400220μmol/L group, donepezil20μmol/L andP38inhibitor SB20358030μmol/L group, donepezil20μmol/L andJNK inhibitor of SP60012520μmol/L group. Each group was givencorresponding inhibitors before30minutes,when donepezil werejoined together. Western blot was used to test APP, ADAM10,ADAM17after24hours; And ELISA was used to detect the levels ofsAPPα, sAPPβ in cellular culture supernatant.Results1.There was on significant difference in the expression of APPprotein among control group,donepezil group, donepezil+LY294002group, donepezil+SB203580group, and donepezil+SP600125group（P＞0.05）. 2.The expression of ADAM10and ADAM17were tested by WB:Compared with the control group, the expressions of ADAM10andADAM17were all increased significantly（P＜0.05）. Compared withthe donepezil group, There was no significant difference in theexpressions of ADAM10and ADAM17in donepezil+LY294002group（P＞0.05）.while the expressions of ADAM10and ADAM17were all decreased significantly in donepezil+SB203580group anddonepezil+SP600125group（P＜0.05）.3.The expressions of sAPPα and sAPPβ were tested byELISA:Compared with the control group, the expressions of sAPPαwere all increased significantly（P＜0.05），while The expressions ofsAPPβ were all decreased. Compared with the donepezil group, therewas on significant difference in the expressions of sAPPαand sAPPβin donepezil+LY294002group（P＞0.05）.while the expressions ofsAPPαwere all decreased and sAPPα were all increased significantlyin donepezil+SB203580group and donepezil+SP600125group（P＜0.05）.ConclusionsDonepezil can make the metabolism of the APP to thenonamyloid way (α metabolic pathways) transition, and themechanisms may involve JNK or P38signal pathways，but may notinvolve PI3-K signal pathways.