Background <p>Neuronal ferroptosis and oxidative stress (OS) critically contribute to epilepsy development, while Tenuigenin (TEN) shows promising neuroprotective effects whose antiepileptic mechanisms warrant further exploration.</p> Objective <p>The study evaluates TEN’s potential to prevent epileptogenesis in immature mice through Nrf2/HO-1 pathway activation, targeting both oxidative stress and ferroptosis as therapeutic mechanisms.</p> Methods <p>A Pentylenetetrazol (PTZ)-induced epileptic model in immature mice was used. After TEN (2, 4, 8&#xa0;mg/kg) administration, the latency and severity of seizures were observed through behavioral tests, and neuronal damage and ferroptosis were assessed using histopathological analysis. OS and ferroptosis markers (ROS, MDA, 4HNE, GSH, Fe²⁺, GPX4, xCT, FTH-1, TfR1), neuroinflammatory cytokines (IL-1β, IL-6, TNF-α, IFN-γ), and apoptosis-related proteins (Bcl2, Bax, Cleaved-caspase3/caspase3) were detected using immunohistochemistry, Immunofluorescence, ELISA, DHE, TUNEL staining, and Western blot techniques. Additionally, the role of TEN in regulating the Nrf2/HO-1 pathway was determined using erastin, oltipraz, and ML385.</p> Results <p>TEN dose-dependently prolonged seizure latency and reduced severity in PTZ-treated immature mice, while decreasing hippocampal neuronal damage and ROS levels. TEN notably downregulated IL-1β, IL-6, TNF-α, IFN-γ, MDA, 4HNE, GSH, Fe²⁺ levels, alleviating neuroinflammation, OS, and ferroptosis. Immunofluorescence and Western blot results revealed that TEN significantly reduced the expression of Bax, cleaved-caspase3/caspase3, and TfR1, while significantly increasing the expression of Bcl2, GPX4, xCT, and FTH-1, reducing neuronal loss and inhibiting ferroptosis. Mechanistic studies found that TEN activated the Nrf2/HO-1 axis by upregulating Nrf2, HO-1, and NQO1, while downregulating Keap1, enhancing antioxidant effects, and inhibiting ferroptosis. Notably, erastin significantly reversed the neuroprotective effects of TEN in epileptic mice.</p> Conclusion <p>TEN protects against epilepsy by activating Nrf2/HO-1, enhancing antioxidants, and inhibiting ferroptosis, suggesting therapeutic potential for pediatric epilepsy.</p> Graphical abstract <p>Tenuigenin exerts neuroprotective effects in immature mice by activating Nrf2/HO-1 signaling, which lowers oxidative stress, strengthens antioxidant capacity, and blocks ferroptosis, thereby reducing epilepsy susceptibility.</p> <p></p> Highlights <p><OrderedList> <ListItem> <ItemNumber>1.</ItemNumber> <ItemContent> <p>Tenuigenin (TEN) alleviates neuronal damage, neuroinflammation, and neuronal loss in epileptic immature mice.</p> </ItemContent> </ListItem> <ListItem> <ItemNumber>2.</ItemNumber> <ItemContent> <p>TEN decreases ferroptosis and oxidative stress in the hippocampus of epileptic immature mice.</p> </ItemContent> </ListItem> <ListItem> <ItemNumber>3.</ItemNumber> <ItemContent> <p>TEN triggers the activation of the Nrf2/HO-1 pathway in the hippocampus of epileptic immature mice.</p> </ItemContent> </ListItem> <ListItem> <ItemNumber>4.</ItemNumber> <ItemContent> <p>Inducing ferroptosis counteracts TEN's neuroprotective effects in the hippocampus of epileptic immature mice.</p> </ItemContent> </ListItem> </OrderedList></p>