<p>Glutamate toxicity is involved in the pathogenesis of neurological disorders, including epilepsy. Here we aimed to investigate the roles of Spermidine/spermine N1-acetyltransferase 1 (SAT1) in regulating glutamate-induced oxidative stress, ferroptosis, and apoptosis in mouse hippocampal HT22 cells. The results indicated that SAT1 expression was upregulated in mice with epilepsy and glutamate-stimulated HT22 cells. Knockdown of SAT1 exhibited antioxidant activity in glutamate-stimulated HT22 cells, as evidenced by increased activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), and decreased reactive oxygen species (ROS) production and malondialdehyde (MDA) content. SAT1 knockdown upregulated the expression of glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11), downregulated acyl-CoA synthetase long-chain family member 4 (ACSL4) expression, and reduced the Fe²⁺ level in glutamate-stimulated HT22 cells, suggesting that SAT1 knockdown reduced ferroptosis in glutamate-stimulated HT22 cells. SAT1 knockdown also decreased the apoptosis rate, caspase-3 activity, and Bcl2-associated X protein (Bax) expression, and increased B-cell leukemia/lymphoma 2 (Bcl-2) expression in glutamate-stimulated HT22 cells. Additionally, SAT1 knockdown reversed the inactivation of nuclear factor-erythroid 2 related factor 2 (Nrf2)/antioxidant response element (ARE) pathway in glutamate-stimulated HT22 cells. Nrf2 knockdown attenuated the effects of SAT1 knockdown on glutamate-induced oxidative stress, ferroptosis, and apoptosis. In conclusion, these findings indicate that SAT1 knockdown prevents glutamate-induced oxidative stress, ferroptosis, and apoptosis in HT22 cells by activating the Nrf2/ARE pathway.</p>