SP1 and ELAVL1 cooperatively regulate ALOX15-mediated ferroptosis in an OGD/R-Induced SK-N-SH cell model of cerebral infarction

Background Cerebral infarction (CI) is associated with high incidence and mortality rates. This study aimed to investigate the role and molecular mechanism of arachidonate 15-lipoxygenase (ALOX15) in oxygen-glucose deprivation/reoxygenation (OGD/R)-induced SK-N-SH cells.Methods An in vitro OGD/R model was established using SK-N-SH cells. The gene expression levels were assessed by RT-qPCR and western blotting. Cell proliferation and apoptosis were evaluated using the 5-Ethynyl-2’-deoxyuridine (EdU) assay and flow cytometry. Ferroptosis-related markers, including reactive oxygen species (ROS), Fe2+, and malondialdehyde (MDA), were measured. Bioinformatics analysis, dual-luciferase reporter, and chromatin immunoprecipitation (ChIP) assays were used to validate the transcriptional regulation of ALOX15 by SP1. The interaction between ELAV-like RNA-binding protein 1 (ELAVL1) and ALOX15 was analyzed using RNA-binding protein database analysis. In vivo, a middle cerebral artery occlusion (MCAO) model was used to assess the effects of ALOX15 silencing on cerebral infarct volume and neurological deficits.Results ALOX15 was upregulated in CI patient tissues and OGD/R-treated SK-N-SH cells. Silencing ALOX15 alleviated OGD/R-induced SK-N-SH cell injury and ferroptosis. Specificity protein 1 (SP1) transcriptionally activated ALOX15 expression, promoting cell damage and ferroptosis. Additionally, ELAVL1 stabilized ALOX15 mRNA, further enhancing its expression and contributing to OGD/R-induced injury and ferroptosis. In the MCAO model, ALOX15 knockdown reduced infarct volume and improved neurological outcomes.Conclusion SP1 and ELAVL1 regulated ALOX15 to drive OGD/R-induced SK-N-SH cell injury and ferroptosis. These findings highlight the SP1/ELAVL1-ALOX15 axis as a potential therapeutic target for mitigating brain injury in CI, providing a translational basis for future interventions.