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PKM2 preconditioning protects endothelial cells from pyroptosis and BBB disruption via NRF2/TRX/TXNIP signaling in neonatal hypoxic-ischemic brain injury
Affiliations
- 1 Department of Neonatology, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 109 Xueyuan West Road, Wenzhou, Zhejiang, 325027, China.
- 2 Key Laboratory of Perinatal Medicine of Wenzhou, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
- 3 Key Laboratory of Structural Malformations in Children of Zhejiang Province, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
- 4 Hongqiao International Institute of Medicine, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China. wxy@shsmu.edu.cn.
- 5 Department of Neonatology, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 109 Xueyuan West Road, Wenzhou, Zhejiang, 325027, China. zhenlanglin@wmu.edu.cn.
- 6 Key Laboratory of Perinatal Medicine of Wenzhou, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China. zhenlanglin@wmu.edu.cn.
- 7 Key Laboratory of Structural Malformations in Children of Zhejiang Province, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China. zhenlanglin@wmu.edu.cn.
- PMID: 41530786
- PMCID: PMC12888557
- DOI: 10.1186/s12974-026-03694-6
Abstract
Background: Neonatal hypoxic-ischemic brain damage (HIBD) is a leading cause of neurological deficits and death in neonates. In HIBD, the death of endothelial cells and disruption of the blood-brain barrier (BBB) are closely related to the severity of brain damage and long-term clinical outcomes. There is increasing evidence that a glycolytic enzyme, pyruvate kinase M2 (PKM2), is essential for managing metabolic processes in endothelial cells, but its role (and underlying molecular mechanism) in hypoxic-ischemic (HI)-associated endothelial cell metabolism, cell survival, and BBB function remains unknown.
Methods: We established an in vivo HI-induced brain injury rat model and an in vitro model in which human cerebral microvascular endothelial cells (hCMECs) underwent oxygen-glucose deprivation (OGD). Infarct volume was measured and neurobehavioral tests were conducted to assess brain damage, and Evans blue extravasation and FITC-dextran were used to evaluate the BBB. RNA sequencing, qRT-PCR, western blotting, and immunofluorescence labeling were conducted to identify the molecular mechanisms underlying HIBD.
Results: PKM2 expression was upregulated in the brains of HIBD rats and in OGD-treated hCMECs. The inhibition of PKM2 greatly upregulated the expression of pyroptosis-associated proteins, including NLRP3, cleaved caspase-1, GSDMD, IL-1?, and IL-18. In contrast, the activation of PKM2 preserved junctional proteins and maintained the integrity of the BBB, which together improved functional recovery in HIBD rats. Mechanistically, preconditioning of PKM2 contributed to lactate-mediated cellular defense mechanisms, including the activation of nuclear factor erythroid 2-related factor 2 (NRF2) and thioredoxin (TRX), and to the downregulation of thioredoxin-interacting protein (TXNIP) via a modest increase in reactive oxygen species.
Conclusions: Our analyses provide compelling evidence that PKM2 preconditioning attenuates endothelial cell pyroptosis and BBB disruption in neonatal HIBD by causing oxidative stress resistance and activating the NRF2/TRX/TXNIP pathway. Therefore, PKM2 represents a promising pharmacological target for treating HIBD.
Keywords: Blood�brain barrier; Hypoxic-ischemic brain damage; Neonate; Pyroptosis; Pyruvate kinase M2.
