Hexokinase 2 upregulation is associated with glycolytic reprogramming and neuroinflammation in hypoxic-ischemic brain damage: a therapeutic target for early intervention.
Hypoxic-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.
Glycolysis-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.
Bioinformatics 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. Early administration of 3-BrPA reduced acute cerebral infarction volume (P < 0.001), improved neurological function scores (P < 0.001), and concurrently downregulated HK2 protein levels (P < 0.05). 3-BrPA treatment also significantly reduced lactate accumulation in the injured brain tissue (P < 0.001). It also suppressed the mRNA expression of pro-inflammatory cytokines (Tnf, Il1b, and Il6; all P < 0.001) and modulated the protein levels of inflammatory markers (iNOS, ARG1; P < 0.05 and P < 0.01, respectively). Moreover, this single early intervention significantly mitigated long-term brain tissue loss and improved motor coordination and exploratory behavior at 30 days post-injury (P < 0.05).
Hk2 is highlighted as a critical node associated with both glycolytic reprogramming and neuroinflammation in HIBD, with upregulation primarily in microglia. Early perturbation of glycolysis with 3-BrPA is associated with multifaceted benefits. Our findings link the Hk2-glycolysis axis to neuroinflammation, offering a rationale for exploring metabolic interventions in HIBD.
Glycolysis-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.
Bioinformatics 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. Early administration of 3-BrPA reduced acute cerebral infarction volume (P < 0.001), improved neurological function scores (P < 0.001), and concurrently downregulated HK2 protein levels (P < 0.05). 3-BrPA treatment also significantly reduced lactate accumulation in the injured brain tissue (P < 0.001). It also suppressed the mRNA expression of pro-inflammatory cytokines (Tnf, Il1b, and Il6; all P < 0.001) and modulated the protein levels of inflammatory markers (iNOS, ARG1; P < 0.05 and P < 0.01, respectively). Moreover, this single early intervention significantly mitigated long-term brain tissue loss and improved motor coordination and exploratory behavior at 30 days post-injury (P < 0.05).
Hk2 is highlighted as a critical node associated with both glycolytic reprogramming and neuroinflammation in HIBD, with upregulation primarily in microglia. Early perturbation of glycolysis with 3-BrPA is associated with multifaceted benefits. Our findings link the Hk2-glycolysis axis to neuroinflammation, offering a rationale for exploring metabolic interventions in HIBD.
Authors
Li Li, Qin Qin, Miao Miao, Mu Mu, Zhang Zhang, Zhang Zhang, Zhao Zhao, Zhang Zhang
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