Identification of Ferroptosis-related Regulatory Network and Validation of the Expression of miRNA-326–IL-1β in Spinal Cord Injury

Abstract

Abstract Background Spinal cord injury (SCI) is a severe central nervous system injury. Emerging research suggests a connection between SCI and ferroptosis. However, its underlying mechanism remains incompletely understood. This study aims to identify key genes associated with ferroptosis after SCI and explore their potential molecular mechanisms. Methods Ferroptosis-related genes (FRGs) were identified by intersecting GSE151371 and the FerrDb database. Enrichment analysis was performed with Gene Ontology (GO) / KEGG. And the top five hub FRGs were from protein-protein interaction network analysis. Subsequently, the competing endogenous RNA (ceRNA) network was constructed based on ENCORI dataset. Furthermore, rat SCI Model was constructed and Basso-Beattie-Bresnahan Locomotor Scale Assessment was used to evaluate hind limb motor function in sham group and SCI group. Ferroptosis marker genes Gpx4, Acsl4 and predicted genes miR-326 – IL-1β were validated through RT-qPCR. Results We screened out 38 FRGs. GO and KEGG analyses revealed that lipid response was significantly associated with ferroptosis after SCI, while IL-17 signaling pathway was predominantly involved in the regulation of ferroptosis. Moreover, we identified five hub FRGs - PPARG, IL-1β, PTGS2, IFNG, and MAPK3 - which played crucial roles in the ceRNA network. Furthermore, the RNA expression level of Acsl4 was upregulated in the SCI group than in the sham group, while the Gpx4 was reversed. Similarly, in comparison to the sham group, the expression level of IL-1β was increased in the SCI group, while miR-326 exhibited a decrease expression. Conclusions miR-326–IL-1β may play pivotal roles in the molecular mechanisms underlying ferroptosis after SCI. Further experimental validation is warranted.