LAUSR

ABSTRACT Changes in mitochondrial structure and function are the initial factors of cell aging. Spermidine has an antiaging effect, but its effect on neuronal aging and mitochondrial mechanisms is unclear. In this study, mouse neuroblastoma (N2a) cells were treated with d‐galactose (d‐Gal) to establish cell aging to investigate the antiaging effect and mechanisms of spermidine. Changes in the cell cycle and &bgr;‐galactosidase activity were analyzed to evaluate the extent of cell aging. Stabilities of mitochondrial mRNA and mitochondrial membrane potential (MMP) were evaluated in the process of cell aging under different treatments. The mitochondrial function was also evaluated using the Seahorse Metabolic Analysis System combined with ATP production. The unfolded protein response (UPR) of the N2a cells was analyzed under different treatments. Results showed that spermidine pretreatment could delay the cell aging and could maintain the mitochondrial stability during d‐Gal treatment. Spermidine increased the proportion of cells in the S phase and maintained the MMP. The oxygen utilization and ATP production in the N2a cells were reduced by d‐Gal treatment but were partially rescued by the spermidine pretreatment. Spermidine ameliorated the N2a cell aging by promoting the autophagy and inhibiting the apoptosis except the UPR. These results showed that spermidine could ameliorate the N2a cell aging by maintaining the mitochondrial mRNA transcription, MMP and oxygen utilization during the d‐Gal treatment. HIGHLIGHTSSpermidine maintained mitochondrial stability of N2a cells following d‐Gal treatment.Spermidine protects N2a cells against the d‐Gal damage by regulating the cell cycle.Spermidine decreased the expression of caspase‐3 and p53 in N2a cells induced by d‐Gal treatmentSpermidine increased the oxygen consumption in N2a cells after d‐Gal treatment.Spermidine enhanced the autophagy in N2a cells following d‐Gal treatment.