Effects of Sirt1 on synaptic vesicle protein expression in SH-SY5Y cells

He Jing; Wang Rong

doi:10.3969/j.issn.1009-6574.2024.04.003

Abstract：Objective To explore the effect of Sirt1 on the expression of synaptic vesicle protein and its function at presynaptic sites.Methods Endogenous Sirt1 expression in SH-SY5Y cells was activated using CRISPRa technology.The cells were divided into aSirt#1,aSirt1#2,aSirt1#3,and aSirt1#NC groups by transfection with aSirt1 plasmid,and a blank control(Mock)group was set up.Lactate dehydrogenase was used to detect cytotoxicity,and Western blot was used to detect the expression levels of synaptic vesicle protein and autophagy related proteins.Results The cytotoxicity of the aSirt1#1 and aSirt1#3 groups was lower than that of the aSirt1#NC group,and the difference was statistically significant(P＜0.05).The expression levels of Sirt1,synaptophysin,vesicle associated membrane protein 2(VAMP2),and alpha-synuclein(α-Syn)in cells of aSirt1#1 group were higher than those of the Mock group,and the difference was statistically significant(P＜0.05).The expression levels of autophagy related proteins Atg13 and Becln1 in SH-SY5Y cells of aSirt1#1 group were higher than those in the Mock group,and the expression levels of autophagy substrate SQSTM1 were lower than those in the Mock group,with statistical differences(all P＜0.05).The P-AMPK/AMPK and P-ULK1/ULK1 ratio in SH-SY5Y cells of aSirt1#1 group were higher than those in the Mock group,and the differences were statistically significant(P＜0.05).Conclusions Sirt1 can regulate autophagy by activating the AMPK-ULK1 pathway,affecting the expression of synaptic vesicle proteins,promoting synaptic vesicle circulation,and affecting the function of presynaptic sites.