Identification of the IL-1β/miRNA-326 axis as a key regulator of ferroptosis in spinal cord injury.

Abstract

INTRODUCTION: Spinal cord injury (SCI) is a devastating condition with high rates of disability and mortality, and secondary injury mechanisms such as ferroptosis have emerged as crucial contributors to its pathophysiology. However, the regulatory mechanisms underlying ferroptosis in SCI remain largely unclear. METHODS: We analyzed peripheral blood transcriptome data (GSE151371) from 10 healthy controls, 10 non-CNS trauma patients, and 38 SCI patients to identify differentially expressed mRNAs (DE-mRNAs) and miRNAs (DE-miRNAs). Ferroptosis-related genes (FRGs) were screened by intersecting DE-mRNAs with a curated list of FRGs. Functional enrichment analyses, regulatory network construction (lncRNA–miRNA–mRNA, TF-miRNA-mRNA), and drug target predictions were conducted. Key findings were validated using the GSE166009 dataset from a rat SCI model and through experimental verification in a TSCI rat model. RESULTS: We identified 38 differentially expressed FRGs (DE-FRGs) in SCI, enriched in lipid metabolism and inflammatory pathways, including IL-17 and TNF signaling. Protein–protein interaction network analysis identified IL-1β as a central hub gene. A miRNA–mRNA network revealed miR-326 as a key regulator of IL-1β. The IL-1β/miR-326 axis was also closely associated with lncRNA-miRNA-mRNA networks, transcription factor regulation, and drug target predictions. Validation in the GSE166009 dataset and a rat SCI model confirmed progressive downregulation of miR-326 and upregulation of IL-1β post-SCI. Dual-luciferase reporter assays demonstrated that miR-326 directly binds the 3′UTR of IL-1β, suppressing its expression. DISCUSSION: Our study highlights the IL-1β/miR-326 axis as a critical regulator of ferroptosis in SCI and a potential therapeutic target for intervention.