Abstract

Background Spinal cord injury (SCI) causes severe energy metabolism dysfunction, hindering neuronal survival and recovery. Adipose tissue-derived stromal cells (ADSCs) have neuroprotective potential, but their role in regulating neuronal energy metabolism and the underlying mechanisms remain unclear. This study aimed to investigate whether ADSCs are capable of restoring neuronal glycolysis through the FOXK1-HK2 signaling pathway, thereby replenishing the energy supply and facilitating tissue regeneration. Methods We employed rat and cell models of SCI to observe the effects of ADSCs on glycolytic metabolism and apoptosis. Transcriptome sequencing identified glycolysis-related differentially expressed genes. Lactate detection and Seahorse assays were used to quantify glycolytic activity. Dual-luciferase reporter assays verified the FOXK1-HK2 regulatory relationship. Cut&Run assay provided direct evidence of FOXK1 binding to the HK2 promoter. Behavioral tests, histopathological staining and immunofluorescence were used to evaluate in vivo functional recovery and tissue repair. FOXK1 knockdown confirmed its role in the ADSC-mediated pathway. Results We found that ADSCs exerted multiple protective and regulatory effects on neurons and motor function. Specifically, they strongly inhibited neuronal oxidative stress, protected mitochondria, and promoted neuronal metabolic reprogramming. Additionally, ADSCs increased glycolytic activity and lactate production, which further contributed to promoting neuronal survival and the recovery of hindlimb motor function. Blocking TGF-β1 signaling abrogated ADSC-induced activation of the FOXK1-HK2 axis and subsequent enhancement of glycolysis, confirming TGF-β1 as a critical paracrine mediator. Through interaction with HK2, FOXK1 plays a critical role in modulating glycolysis. Dual-luciferase reporter and Cut&Run assays confirmed that FOXK1 regulates the HK2 promoter, thereby increasing its transcriptional activity.