Dinotefuran aggravates Alzheimer's disease-associated microglial inflammation through the CHUK/NFKB1-mediated NF-κB signaling pathway

Neonicotinoid pesticides are widely used systemic insecticides with increasing evidence of neurotoxicity. However, their potential contribution to Alzheimer's disease (AD)-related neuroinflammation remains poorly understood. In this study, seven representative neonicotinoid pesticides were systematically evaluated by integrating physicochemical characterization, target prediction, pathway enrichment analysis, human AD snRNA-seq, molecular docking, molecular dynamics simulations, and experimental validation. Among the seven compounds, dinotefuran was prioritized based on its favorable blood-brain barrier permeability and enrichment in inflammation-related pathways. Integration of target prediction with AD transcriptomic analyses identified CHUK and NFKB1 as key candidate targets associated with NF-κB signaling. snRNA-seq analysis further demonstrated preferential enrichment of NF-κB signaling activity in microglia from AD brains. Molecular docking and molecular dynamics simulations supported potential interactions between dinotefuran and IKKα (CHUK) as well as NF-κB1 (p50). In Aβ-treated BV2 microglia, dinotefuran further enhanced activation of the CHUK/NFKB1-mediated NF-κB signaling pathway, accompanied by increased expression of IKKα, phosphorylation of IκBα and p65, enhanced NF-κB1 activation, and elevated secretion of IL-1β, TNF-α, IL-6, IFN-α, and IFN-β. These pro-inflammatory effects were markedly attenuated by the NF-κB inhibitor BAY11-7082. Collectively, our findings suggest that CHUK/NFKB1-associated NF-κB signaling may contribute to dinotefuran-enhanced inflammatory responses in Aβ-treated BV2 microglia, providing preliminary mechanistic insights into the potential involvement of neonicotinoid pesticides in AD-related neuroinflammation.