The Mechanism of Long Non-coding RNA GABPB1-AS1/SLC12A5 Regulating Neuronal Ferroptosis in Epileptic Seizures

Epilepsy, a common neurological disorder, is distinguished by abnormal neuronal activity. This study explored the mechanism of the long non-coding RNA (lncRNA) GABPB1-AS1/SLC12A5 pathway modulating neuronal ferroptosis during epileptic seizures. Loss- and gain-of-function approaches were conducted in a mouse model of kainic acid-induced seizures and glutamic acid (Glu)-treated HT22 neurons to explore the regulatory effect of lncRNA GABPB1-AS1/SLC12A5 on neuronal ferroptosis. Mouse seizures were assessed using the Racine scale. LncRNA GABPB1-AS1, SLC12A5, and ferroptosis-related markers were measured via RNA quantification, Western blot, and immunofluorescence. The direct or indirect interaction between lncRNA GABPB1-AS1 and SLC12A5 was validated using RIP, RNA pull-down, and Co-IP assays. CHX chase and ubiquitination assays assessed SLC12A5 stability and ubiquitination levels regulated by lncRNA GABPB1-AS1. LncRNA GABPB1-AS1 was upregulated in the hippocampal CA1 region of KA-induced mice. LncRNA GABPB1-AS1 knockdown suppressed neuronal ferroptosis-lowering Fe²⁺, MDA, and lipid peroxidation, while raising GSH, SLC7A11, and GPX4. Its knockdown also reduced seizure scores and frequency, prolonged seizure latency, and shortened seizure duration. An in vitro experiment verified the suppressive effect of lncRNA GABPB1-AS1 knockdown on neuronal ferroptosis. In addition to a direct binding relationship, this lncRNA further recruited the E3 ligase PRKN to enhance SLC12A5 ubiquitination and proteasomal degradation. SLC12A5 overexpression inhibited Glu-induced ferroptosis, and SLC12A5 knockdown partially reversed the effects exerted by lncRNA GABPB1-AS1. LncRNA GABPB1-AS1 recruits the E3 ligase PRKN to promote ubiquitination and proteasomal degradation of SLC12A5, thereby downregulating SLC12A5 expression and facilitating hippocampal neuronal ferroptosis that mediates epileptic seizures.