ObjectiveTo screen autophagy-related hub genes in renal ischemia-reperfusion injury (RIRI) and analyze their biological functions to investigate the pathogenesis of RIRI.MethodsTwelve ICR mice were randomly assigned to a control group and an RIRI group (n=6 per group). Renal tissues from both groups were subjected to transcriptome sequencing. After quality control filtering, differentially expressed genes (DEGs) between the two groups were identified. The intersection of these DEGs with autophagy-related genes (ATGs) yielded autophagy-related DEGs (ATG-DEGs). Hub genes were selected as the top-ranked ATG-DEGs using four algorithms in Cytohubba. Single-gene enrichment analysis was performed for the hub genes. Two murine RIRI-related gene sequencing datasets were downloaded from the NCBI GEO database for single-cell RNA-seq analysis to identify key genes at the single-cell level, followed by immune infiltration analysis. Finally, experimental validation of hub gene expression levels was conducted.ResultsA total of 672 DEGs were identified between the two groups. GO and KEGG enrichment analyses suggested that these DEGs might be associated with autophagy. Six hub genes were identified: phagocytic glycoprotein-1 (Cd44), integrin subunit alpha M (Itgam), complement component 3 receptor 4 subunit (Itgax), kinase insert domain receptor (Kdr), Secreted phosphoprotein 1 (Spp1), and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH). These genes were involved in signaling pathways related to oxidative stress, inflammatory response, apoptosis, and immune response. Based on the murine RIRI-related data downloaded from the GEO database, single-cell transcriptome analysis revealed that two hub genes (Cd44 and Itgam) were highly expressed in neutrophils, macrophages, and fibroblasts in the RIRI group compared to the control group (P<0.05). Cd44⁺ neutrophils and Itgam⁺ macrophages exhibited more significant cell communication. Immune infiltration results showed higher correlations of Cd44 and Itgam with neutrophils and macrophages in the RIRI group than that in control group. qRT-PCR results confirmed that the expression levels of Cd44 and Itgam were significantly higher in RIRI group than that in control group (P<0.05).ConclusionCd44 and Itgam are autophagy-related hub genes in murine RIRI and may serve as novel biomarkers for prevention and treatment of RIRI.

renal ischemia-reperfusion injury;bioinformatics analysis;hub genes;phagocytic glycoprotein-1;integrin subunit alpha M