Molecular mechanisms of ferroptosis in renal ischemia-reperfusion injury Investigated via bioinformatics analysis and animal experiments

Kidney transplantation is a pivotal treatment for end-stage renal disease. However, renal ischemia-reperfusion injury (IRI) during surgery significantly impacts graft function. Despite unclear molecular mechanisms, no specific therapies or preventative measures are available. Gene expression profiles from renal biopsies before and after IRI were downloaded from public databases. Differentially expressed genes were identified using the Wilcoxon rank-sum test and weighted gene co-expression network analysis. Ferroptosis-associated genes were screened using the FerrDb database. The genes with the highest connectivity were identified via the protein-protein interaction (PPI) network and upstream regulatory miRNAs were found through the gene-miRNA network. A mouse renal IRI model was constructed for transcriptome sequencing and quantitative real-time polymerase chain reaction (qRT-PCR) validation to elucidate the relationship between key ferroptosis genes and regulatory miRNAs in renal IRI. Differential analysis identified 15 ferroptosis-associated genes (<i>TNFAIP3</i>, <i>IL6</i>, <i>KLF2</i>, <i>EGR1</i>, <i>JUN</i>, <i>ZFP36</i>, <i>GDF15</i>, <i>CDKN1A</i>, <i>HSPB1</i>, <i>BRD2</i>, <i>PDK4</i>, <i>DUSP1</i>, <i>SLC2A3</i>, <i>DDIT3</i>, and <i>CXCL2</i>) involved in renal IRI regulation. In animal experiments, ferroptosis-related genes were also upregulated in the model group. Enrichment analysis and hematoxylin-eosin pathological staining suggested these genes are primarily involved in renal inflammatory responses. PPI network analysis revealed <i>IL6</i> as the gene with the highest connectivity, and the gene-miRNA network indicated <i>IL6</i> might be regulated by <i>miR-let-7a.</i> Animal experiments revealed decreased <i>miR-let-7a</i> and increased <i>IL6</i> levels in the model group, identifying potential therapeutic targets. <i>MiR-let-7a</i> regulates ferroptosis in renal IRI by targeting <i>IL6</i>, highlighting <i>IL6</i> as a crucial gene in the ferroptosis process of renal IRI.