Unveiling the neuroinflammatory potential of aspartame in Alzheimer’s disease: A comprehensive network toxicology and experimental validation approach

Aspartame, a widely used artificial sweetener, has raised significant concerns regarding its safety, especially in light of the rising incidence of Alzheimer’s Disease (AD). However, systematic investigations into aspartame’s potential neurotoxicity and its role in AD pathology remain limited. In this study, we employed an integrated approach combining network toxicology predictions, molecular docking, and in vitro validation to explore the neurotoxic mechanisms of aspartame in AD. By analyzing multiple public databases through network toxicology, we identified 185 common targets between aspartame and AD. Protein-protein interaction (PPI) network analysis further highlighted 10 core hub genes, including KDR, CCNE1, and GSK-3β. Molecular docking predicted favorable interactions for key targets such as GSK-3β (−5.002 kcal/mol) and KDR (−5.613 kcal/mol), with other hubs showing moderate binding affinities. In vitro experiments demonstrated that aspartame dose-dependently reduced BV2 microglial cell viability and triggered a pro-inflammatory response. Specifically, aspartame increased the production of nitric oxide (NO) and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6), while upregulating the expression of GSK-3β, iNOS, TNF-α, IL-1β, and IL-6, similar to LPS-induced inflammation. These findings suggest that aspartame may induce a pro-inflammatory response in BV2 microglial cells through multiple signaling pathways, although the relevance of these findings to human neurological safety remains to be established. This study provides a basis for further investigation into the neurological safety of aspartame and proposes a novel paradigm for evaluating the neurotoxic potential of food additives using an integrated systems toxicology approach.