RAF1 Promotes Gstt1‐Mediated Ferroptosis by Promoting KDM1A Recruitment to Facilitate Pilocarpine‐Induced Acute Seizure‐Associated Neuronal Injury

ABSTRACT Background Seizures can cause progressive neuronal damage. Our previous studies found that RAF1 mediates vitamin D's alleviation of seizures, but the mechanism by which RAF1 acts in seizures remains unclear. Methods The pilocarpine‐induced acute seizure‐associated neuronal injury (Pilo‐ASNI) model and glutamate‐treated HT22 cells were used. Behavioral assessments were performed to evaluate seizure severity and latency; histopathological staining was performed to examine neuronal damage and survival; and cellular assays were performed to assess cell viability, cytotoxicity, and DNA damage. Transcriptome sequencing, Co‐immunoprecipitation, molecular docking, ChIP‐qPCR, and ferroptosis indicators were employed to elucidate the molecular mechanism. Results RAF1 expression was markedly upregulated in the Pilo‐ASNI model and glutamate‐treated HT22 cells. RAF1 knockdown attenuated seizure severity and neuronal damage. Mechanistic studies in HT22 cells revealed that RAF1 inhibited Gstt1 transcription and promoted ferroptosis by promoting KDM1A recruitment and reducing H3K4me2 levels in the promoter region of Gstt1. Disrupting the RAF1‐KDM1A interaction could inhibit ferroptosis and alleviate nerve damage, whereas Gstt1 knockdown reverses the protective effect of RAF1 inhibition. Conclusion This study revealed a RAF1‐driven pathway involving KDM1A‐mediated epigenetic suppression of Gstt1 and subsequent ferroptosis, which may represent a potential target for neuroprotective intervention in seizures.