Network toxicology‑guided validation of glyphosate’s neurotoxic mechanisms in human: From in silico prediction to experimental proof

Glyphosate, a widely used herbicide, has raised increasing concerns regarding its potential neurotoxic effects in humans. This study aimed to elucidate the underlying mechanisms of glyphosate-induced neurotoxicity by integrating network toxicology, molecular docking, and experimental validation. Using publicly available databases, we identified human-specific targets of glyphosate and overlapped them with neurotoxicity-associated genes to construct a protein-protein interaction (PPI) network. Analysis identified critical hub genes SRC, TNF, and CASP3, key regulators implicated in synaptic dysregulation, neuroinflammation, and apoptosis, respectively. Molecular docking revealed stable binding affinities between glyphosate and these targets. Experimental validation in BV2 microglial cells exposed to environmentally relevant glyphosate concentrations (0.5-5 mg/L) for 72 h confirmed increased phosphorylation of Src (Tyr419), elevated TNF-α, and cleaved caspase-3 expression. These results highlight Src as a previously unrecognized mediator of glyphosate-induced neurotoxicity and demonstrate the value of a combined in silico-in vitro strategy in environmental neurotoxicology.