Benzene metabolite catechol promotes microglia-mediated neuroinflammation in Alzheimer's disease via regulating NF-κB signaling.

Benzene is an occupational and environmental toxicant, and its exposure is associated with increased risk of Alzheimer's disease (AD), yet its underlying mechanisms, particularly the roles of benzene metabolites, have not been elucidated. Here, benzene and its major metabolites were systematically evaluated through physicochemical characterization, blood-brain barrier permeability prediction, target identification, and pathway enrichment analysis. Single-nucleus RNA sequencing data were further analyzed using pathway activity scoring approaches to assess cell-type-specific signaling patterns. Molecular docking and molecular dynamics simulations were performed to explore interactions between selected metabolites and key regulatory proteins, and in vitro experiments were conducted for validation. Among the analyzed compounds, catechol was prioritized based on its predicted central nervous system accessibility and strong association with AD-related pathways. Pathway enrichment and transcriptomic scoring consistently highlighted NF-κB signaling as a key pathway, with preferential activation in microglia. Mechanistically, RELA (p65), a central regulator of NF-κB signaling, was identified as a potential target, and structural as well as experimental analyses supported the interaction between catechol and p65. Functional validation demonstrated that catechol enhanced Aβ-induced microglial activation and inflammatory responses. Collectively, these findings suggest that catechol may act as a critical mediator linking benzene exposure to microglia-driven neuroinflammation in AD, providing mechanistic insight into the contribution of environmental pollutants to neurodegenerative disease progression.