Agnuside enhances neuronal survival and cognitive function by modulating PI3K/AKT/mTOR signaling in Aluminium...

Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder characterized by chronic inflammation, oxidative stress, and amyloid beta (Aβ) aggregation. Prolonged exposure to heavy metals such as aluminium (Al) promotes reactive oxygen species (ROS) generation and neuroinflammation, contributing to Aβ aggregation, neuronal damage, and cognitive decline. Although conventional drugs show therapeutic potential, their use is often limited by adverse side effects. Phytochemicals with antioxidant, anti-inflammatory, and anticholinesterase properties are safer alternatives. This study explored the neuroprotective potential of agnuside (AGN) against aluminium chloride (AlCl₃)-induced AD pathology through network pharmacology and experimental approaches. A total of 108 overlapping targets of AGN and AD were identified, with key nodes including TNF, AKT1, ALB, EGFR, ESR1, CASP3, MMP9, SRC, HSP90AA1, and PPARG. Gene ontology and KEGG pathway analyses suggested that AGN may modulate key molecular pathways implicated in AD, including TNF signaling and the PI3K/AKT signaling. Molecular docking confirmed that AGN has strong binding affinities with TNF, AKT1, ALB, MMP9, and CASP3. SH-SY5Y cells and zebrafish larvae exposed to AlCl₃ exhibited AD pathology, which was notably attenuated by AGN, as indicated by reductions in oxidative stress, apoptosis, and neuroinflammation, confirmed through acridine orange (AO), reactive oxygen species (ROS), and neutral red staining. This protective effect was further validated by enzymatic assays and gene expression analysis, confirming its antioxidant and anti-apoptotic potential. These findings suggest that AGN is a promising phytotherapeutic candidate for counteracting AlCl₃-induced neurotoxicity and could serve as a potential therapeutic agent for AD intervention.