Melatonin exerts an in vitro anti‑glioblastoma effect through SIRT1‑mediated HMGB1 deacetylation and inhibition of the TLR4/NF‑κB pathway

functional assays. Network pharmacology, protein-protein interaction (PPI) analysis and molecular docking were performed to identify potential targets and pathways. Western blotting and EX527-mediated SIRT1 inhibition were used to evaluate the involvement of the SIRT1/HMGB1/TLR4/NF-κB axis. MT notably inhibited GBM cell viability, migration and invasion while inducing apoptosis. Network pharmacology and PPI analyses indicated that SIRT1, TLR4 and NF-κB1 were functionally relevant targets, and enrichment analysis suggested the involvement of TLR and NF-κB signaling pathways. Molecular docking further suggested a potential interaction between MT and SIRT1. Mechanistically, MT upregulated SIRT1 protein expression, reduced HMGB1 acetylation, decreased TLR4 expression and suppressed p65 phosphorylation. Conversely, EX527-mediated inhibition of SIRT1 induced molecular changes opposite to those observed after MT treatment, including increased HMGB1 acetylation, TLR4 expression and p65 phosphorylation. The present study suggests that MT suppresses the malignant phenotype of GBM cells, at least partly, in association with increased SIRT1 expression, reduced HMGB1 acetylation and attenuation of TLR4/NF-κB-related signaling. The present findings provide new theoretical support for developing MT as a potential adjuvant therapeutic candidate for GBM.