Network pharmacological combined with in vivo and in vitro validation reveals the mechanisms of atrazine-induced Parkinson's disease-related neurotoxicity.

Atrazine (ATR), a common triazine herbicide, is closely linked to neuroinflammation, yet the molecular mechanism underlying its neurotoxic potential remains poorly defined. This work explored ATR-provoked neuroinflammation focusing on microglial activation and the PI3K/AKT/JNK cascade via network pharmacology, molecular simulation and multilevel experiments. Potential ATR-PD intersecting targets were screened for PPI, GO and KEGG enrichment. Molecular docking and 100 ns dynamics simulations verified stable binding between ATR and hub molecules. ATR-triggered microglial activation, proinflammatory cytokine release and PI3K/AKT/JNK phosphorylation were detected in BV2 cells, which were reversed by AKT inhibitor MK2206. Transwell coculture and conditioned medium assays confirmed that ATR-stimulated microglia caused HT22 neuronal synaptic damage, rescued by MK2206. In chronically ATR-exposed C57BL/6 mice, motor dysfunction, substantia nigra lesion, microgliosis, pathway hyperactivation and neuronal loss (NeuN-positive neurons) were observed. Collectively, ATR facilitates microglia-dependent neuroinflammation and nigral neuronal damage by perturbing PI3K/AKT/JNK signaling. AKT suppression alleviates ATR-mediated neurotoxicity under the present experimental conditions. Notably, the in vitro (40 μM) and in vivo (25 mg/kg/d) doses used in this study are substantially higher than typical human environmental exposure levels; these findings cannot be directly extrapolated to human health risk assessment. This study provides a candidate intervention strategy for ATR-associated neuronal injury observed in the experimental model.