Integrated Experimental and In-Silico Analysis Reveals Melittin as a Modulator of IL-37a-Associated Networks in Neuroblastoma

Background: Neuroinflammation is a key process shaping tumor progression and cellular stress responses. Pro-inflammatory signaling components amplify these responses, whereas IL-37a functions as an endogenous suppressor that limits inflammation. Melittin is a bioactive peptide with cytotoxic and immunomodulatory properties; however, its effects on IL-37a-centered immune regulation in neuroblastoma cells remain unclear. Methods: Human SH-SY5Y neuroblastoma cells were exposed in vitro to melittin at concentrations of 1–32 µM for 24 and 48 h. Cytotoxicity was assessed using the MTT assay. Transcriptional changes in IL-37a-associated signaling components were quantified by qRT-PCR. Pathway-level interactions were analyzed using STRING-based protein–protein interaction networks, Gene Ontology, and KEGG pathway enrichment. The binding potential of melittin to STAT3 and IL-1R8 was evaluated by molecular docking, and in silico ADMET properties were predicted. Results: Melittin induced dose- and time-dependent cytotoxicity in SH-SY5Y cells, with an IC₅₀ of 16 µM at 24 h. Melittin significantly increased IL-37a, IL-18R1, and IL-1R8 expression while suppressing NF-κB, STAT3, MyD88, and Smad3. Caspase-1 expression was elevated, indicating modulation of inflammation-associated cell death pathways. Extracellular matrix remodeling was differentially regulated, with upregulation of MMP-2 and downregulation of MMP-9. Multivariate analyses revealed a treatment-specific transcriptional signature, and enrichment analyses highlighted IL-1 family, Toll-like receptor, and JAK–STAT signaling pathways. Docking analyses supported high-affinity binding of melittin to IL-1R8 and STAT3, while ADMET profiling suggested limited permeability and low predicted cardiotoxicity. Conclusion: Melittin exerts a multilayered immunomodulatory effect in SH-SY5Y neuroblastoma cells by activating an IL-37a-centered anti-inflammatory response while suppressing canonical pro-inflammatory signaling pathways.