<p>Alzheimer’s disease (AD) is a progressive neurodegenerative disorder associated with memory loss, spatial disorientation, and a marked decline in cognitive capacity. Colchicine, a microtubule-disrupting agent, is widely used to model cognitive dysfunction in animals. This study aimed to assess the therapeutic potential of Diosgenin against AD utilizing an integrated in silico, in vitro, and in vivo approach. The in silico analyses, molecular docking and molecular dynamics simulations demonstrated strong binding affinity of Diosgenin with key AD-associated proteins LRP5, MME, and NOS2, indicating modulation of oxidative stress, apoptosis, and neuroinflammation. Computational studies also indicated favourable pharmacokinetic properties, supporting its blood-brain barrier permeability. In vitro assays in BV2 microglial cells demonstrated the antioxidant potential of Diosgenin by reducing oxidative stress markers such as MDA while preserving key antioxidant enzymes SOD and GSH. In vivo studies in a colchicine-induced rat model of AD showed that Diosgenin significantly improved cognitive and memory functions, as evidenced by enhanced performance in radial arm maze and novel object recognition tasks. Brain tissue analysis showed that diosgenin improved cholinergic function by lowering AChE and BChE activity. It also enhanced the brain’s antioxidant defence (SOD and GSH) and reduced lipid peroxidation (MDA), thereby limiting oxidative stress. Microscopic studies further confirmed fewer degenerating neurons, reduced plaques, and less inflammation. Taken together, these findings suggest that diosgenin offers multi-faceted protection in AD-supporting memory-related neurotransmission, reducing oxidative damage, and dampening inflammation. Nevertheless, further investigation into advanced formulation strategies to overcome its limited bioavailability is warranted to enable clinical translation.</p> Graphical abstract <p></p>