Exploring key genes associated with metabolic cell death in ischemic stroke: a comprehensive analysis of single-cell, bulk transcriptome, and experimental validation

Metabolic cell death and ischemic stroke (IS) may be closely correlated. In this study, we identified key metabolic cell death-related genes (MCD-RGs) in IS, which may provide insight into ischemic injury and candidate targets for future therapeutic exploration. GSE163654, GSE30655, and GSE174574 were retrieved from the gene expression omnibus (GEO) database, and MCD-RGs were integrated from the literature. Key genes were screened using expression validation. Nomogram construction, gene set enrichment analysis, key cell identification, and pseudo-time analysis were conducted. Finally, the expression profiles of the key genes were evaluated by RT-qPCR and western blot analyses. Five genes (Atf3, Cdkn1a, Hmox1, Jun, and Ptgs2) involved in metabolic cell death were identified. The nomogram model containing these genes exhibited favorable diagnostic performance in predicting the incidence of IS. These genes were primarily associated with Myc targeting V1, fatty acid metabolism, protein secretion, oxidative phosphorylation, and are involved in TNF-α signaling pathways through NF-κB and peroxisome, G2M checkpoint, glycolysis, and epithelial-mesenchymal transformation. Neutrophils, endothelial cells, and perivascular fibroblast-like cells are key cells. Cdkn1a and Jun expression exhibited dynamic changes along the pseudo-time trajectories of key cells. Their expression was higher in the IS samples, particularly at later pseudo-time states. RT-qPCR and western blot analysis indicated that Cdkn1a, Jun, and Ptgs2 were significantly upregulated in IS samples, whereas Atf3 and Hmox1 showed no significant trend, which was consistent with the transcriptomic results. Five key genes and three key cell types are associated with metabolic cell death in IS were identified, providing candidate molecular and cellular insight for future mechanistic studies and therapeutic exploration.