Hexokinase 2 upregulation is associated with glycolytic reprogramming and neuroinflammation in hypoxic-ischemic brain damage: a therapeutic target for early intervention

BackgroundHypoxic-ischemic brain damage (HIBD) involves profound metabolic reprogramming, where aberrant glycolysis links to neuronal injury. This study aimed to identify and characterize glycolysis-related hub genes in HIBD.MethodsGlycolysis-related differentially expressed genes (DEGs) were screened from the HIBD dataset GSE144456 by combining differential expression analysis, weighted gene co-expression network analysis (WGCNA), and a glycolysis gene set.

Hub genes were further identified via enrichment and protein-protein interaction (PPI) network analyses and validated in an independent dataset (GSE23317) and our internal RNA-seq. The key hub gene was confirmed in a mouse HIBD model using RT-qPCR and Western blot.

To perturb Hexokinase 2 (Hk2), 3-bromopyruvate (3-BrPA, 1 mg/kg) was administered 1 hour post-HIBD, and its effects were evaluated by MRI, molecular assays, and behavioral tests.ResultsBioinformatics analysis identified Hk2 as a key hub gene that was consistently upregulated in HIBD. In vivo experiments demonstrated that both HK2 protein and mRNA levels were significantly elevated 24 hours after HIBD (P < 0.001).

Double immunofluorescence staining further revealed that the upregulated HK2 was predominantly localized to Iba1+ microglia.