<p>Microsomal glutathione S-transferase 1 (MGST1) regulates ferroptosis in cancers, but its engagement in anesthesia-induced neuronal ferroptosis is unclear. This study intended to investigate the influence of MGST1 on ferroptosis in sevoflurane (Sev)-treated HT22 cells. Cells were treated with 1%, 2%, and 4% Sev. HT22 cells were transfected with MGST1 overexpression or negative control plasmids, followed by 4% Sev treatment. Cell viability, oxidative stress markers, iron levels, ferroptosis-related proteins, Wnt pathway, and MGST1 gene expression and protein level were detected. Sev enhanced ferroptosis and reduced MGST1 expression in HT22 cells in a concentration-dependent manner. MGST1 enhanced viability in Sev-treated HT22 cells (<i>P</i> &lt; 0.05). MGST1 reduced reactive oxygen species (ROS) level (<i>P</i> &lt; 0.001) and malondialdehyde (MDA) (<i>P</i> &lt; 0.01), but increased superoxide dismutase (SOD) activity (<i>P</i> &lt; 0.05), indicating that MGST1 inhibited oxidative stress in Sev-treated HT22 cells. MGST1 also reduced Fe²⁺ level in Sev-treated HT22 cells (<i>P</i> &lt; 0.05). Regarding ferroptosis-related proteins, MGST1 increased glutathione peroxidase 4 (GPX4) (<i>P</i> &lt; 0.05) and solute carrier family 7, member 11 (SLC7A11) (<i>P</i> &lt; 0.01) in Sev-treated HT22 cells. Importantly, MGST1 also elevated phosphorylated-glycogen synthase kinase-3 beta (GSK-3β)/GSK-3β (<i>P</i> &lt; 0.01) and β-catenin/β-actin (<i>P</i> &lt; 0.05), which indicated that MGST1 activated the Wnt pathway in Sev-treated HT22 cells. MGST1 suppresses ferroptosis and activates the Wnt pathway in Sev-treated HT22 cells.</p>