Network Pharmacology Combined With Clinical Retrospective Analysis to Investigate the Potential Mechanisms and...

Folate is an indispensable nutrient involved in key biological processes, including enzymatic reactions, DNA replication, metabolic regulation, and methylation. Studies indicate that folate promotes neuronal regeneration and repair in patients with spinal cord injury (SCI); however, its precise mechanisms remain unclear. We employed network pharmacology to identify potential targets of folate for SCI treatment; we conducted a clinical retrospective study, selecting 50 SCI patients and 50 non-SCI control subjects from Tianjin Hospital between 2022 and 2025 to validate predictions by assessing serum folate levels. Network pharmacology identified 1402 folate-related targets and 548 SCI-related targets. Key targets included TNF-α, CASP3, EGF, IL1β, and AKT1. Molecular docking revealed the highest binding affinity between folate and CASP3/TNF-α (-8.5 kcal/mol). Clinical validation demonstrated statistically significant lower folate levels in SCI patients compared to normal levels in non-SCI controls. A strong inverse correlation was observed between folate levels and injury level (r = -0.58, p < 0.001). Folate exerts neuroprotective effects by synergistically regulating neuronal apoptosis, inflammatory responses, and oxidative stress pathways. Clinical data reveal prevalent folate deficiency in SCI patients, and this nutritional deficit may exacerbate secondary injury cascades. We recommend incorporating folate supplementation into comprehensive SCI management protocols.