The miR-30c-5p/SOCS3 axis is a potential driver of inflammation and metabolic imbalance in Duchenne muscular dystrophy.

Duchenne muscular dystrophy (DMD) constitutes a severe, incurable disorder inherited in an X-linked manner, characterized by continuous skeletal muscle degeneration, chronic inflammatory responses, and profound metabolic alterations. Although miRNA-mRNA regulatory networks are thought to contribute to DMD pathogenesis, their key drivers remain insufficiently defined. In this study, we integrated bulk RNA-seq (GSE38417, GSE109178), small RNA-seq (GSE157668), and single-cell RNA-seq (GSE213925) data and combined differential expression analysis, target gene prediction, functional pathway enrichment mapping, and protein-protein interaction network analysis to screen candidate miRNA-mRNA axes, which were subsequently validated in mdx mice, C2C12 myoblasts, and primary skeletal muscle cells. We identified 100 differentially expressed miRNAs (DEMs) in DMD muscle, with miR-30c-5p being the most downregulated and possessing the highest number of predicted targets. Integrated analysis revealed SOCS3 as a key upregulated hub gene targeted by miR-30c-5p. ScRNA-seq showed elevated SOCS3 expression in myocytes from DMD muscle, where SOCS3+ cells exhibited enriched inflammatory pathways and suppressed metabolic processes. In mdx mice, miR-30c-5p expression showed a pronounced reduction (P < 0.001). In contrast, SOCS3 expression increased at both the transcriptional (P < 0.001) and translational (P < 0.01) levels. Functional experiments further showed that overexpression of miR-30c-5p reduced SOCS3 expression, whereas inhibition of miR-30c-5p increased SOCS3 levels in C2C12 cells and primary skeletal muscle cells. Dual-luciferase reporter assay further supported direct binding of miR-30c-5p to the SOCS3 3'UTR. In mdx-derived primary skeletal muscle cells, miR-30c-5p restoration was also accompanied by reduced TNF-α and increased IL-10 levels, and rescue experiments supported that these anti-inflammatory effects were at least partly SOCS3-dependent. These findings suggest that the miR-30c-5p/SOCS3 axis is associated with inflammation- and metabolism-related alterations in DMD and warrants further investigation.