Mechanistic study on ligustilide modulation of the TLR4/NF-κB pathway in ameliorating Scopolamine-Induced cognitive impairment

This research employed a multi-method strategy integrating network pharmacology, molecular docking, and animal experiments to investigate the therapeutic mechanisms of ligustilide in Alzheimer's disease (AD) management. AD-related targets were obtained from GeneCards, while ligustilide-associated targets were identified using SwissTargetPrediction. Protein interaction networks were constructed via the STRING database. Functional annotation and pathway enrichment analysis were performed using Metascape, including Gene Ontology and KEGG pathway analyses revealing that ligustilide modulated AD progression primarily by regulating the Toll-like Receptor 4/nuclear factor kappa B (TLR4/NF-κB) signaling pathway. Molecular docking studies conducted with AutoDock Vina demonstrated strong binding affinities between ligustilide and key targets in this pathway. Behavioral evaluations, including locomotor activity assessment (open field test), spatial memory analysis (Morris water maze), and recognition memory testing (novel object recognition test) demonstrated that ligustilide significantly attenuated SCOP-induced cognitive deficits. Histopathological and biochemical analyses indicated that ligustilide maintained cholinergic neuron integrity and boosted antioxidant defenses in SCOP-treated mice. Transcriptional profiling revealed decreased expression of TLR4, NF-κB p65, and pro-inflammatory mediators (TNF-α, IL-1β, IL-6) in ligustilide-treated AD model mice. Western blot and enzyme-linked immunosorbent assay (ELISA) demonstrated reduced levels of TLR4, phosphorylated p65 (Ser536), TNF-α, IL-1β and IL-6 in ligustilide-treated AD model mice. These findings collectively demonstrate that ligustilide alleviates SCOP-induced cognitive deficits through suppression of the TLR4/NF-κB signaling cascade.

Alzheimer's disease; Cholinergic dysfunction; Inflammatory response; Ligustilide; Scopolamine; TLR4/NF-κB signaling pathway.