Identification of mitochondrial dysfunction-related biomarkers and immune infiltration in liver ischemia-reperfusion injury via integrated bioinformatics and machine learning.

Mitochondrial dysfunction contributes to the pathogenesis of multiple diseases. This study explores the involvement of mitochondrial dysfunction-related genes (MDRGs) in liver ischemia-reperfusion injury (LIRI). Differentially expressed MDRGs (DE-MDRGs) were identified using public databases. We identified key hub genes by integrating functional enrichment, protein-protein interaction (PPI) network analysis, and machine learning algorithms. These candidates underwent rigorous validation using independent cohorts, an in vivo LIRI mouse model, and computational approaches including molecular docking and dynamics simulations. Twelve DE-MDRGs were identified, enriched in autophagy regulation, autophagosome formation, cytokine activity, and pathways linked to neurodegenerative and graft-versus-host diseases. The MCC algorithm prioritized ten DE-MDRGs, with IL-6, HTT, and SLC19A2 consistently selected across machine learning models. ROC analysis confirmed their diagnostic accuracy. A nomogram incorporating these genes demonstrated strong predictive performance for LIRI risk. Immune profiling revealed that LIRI is characterized by an elevated presence of neutrophils, naive CD4 T cells, and activated mast cells, contrasting with a significant reduction in M2 macrophages and resting populations of mast cells and memory CD4 T cells. GSEA further revealed significant enrichments of IL-6, HTT, and SLC19A2 in pathways associated with immune responses. The differential expression of these three genes was further confirmed both in an external validation cohort and in vivo in liver tissues from the murine LIRI model. In silico screening via docking and dynamic simulations suggested that the hub genes could be effectively targeted by Curcumin, Valproic Acid, or Acetylcysteine, all of which displayed promising interaction profiles. Our research uncovered an interaction between mitochondrial dysfunction and the immune microenvironment in LIRI. This finding could offer significant perspectives for the clinical management of LIRI through the regulation of mitochondrial activity.