Lately, interest surrounding the utilization of plant-derived compounds as a viable beneficial approach for treating Alzheimer’s disease (AD) has significantly increased. This study aimed to assess the defensive properties of rosavin against Alzheimer’s disease induced by amyloid-β, utilizing experimental models. We found that rosavin exhibited anti-aggregation and disaggregation properties, suggesting its potential to prevent the gathering of Aβ-aggregates. In vitro experiments revealed that rosavin effectively mitigated the neurotoxicity induced by Aβ in Neuro-2a cells, showcasing its protective potential. Rosavin significantly improved the Aβ-induced cognitive deficits in Wistar rats, particularly in spatial memory. Which the pathophysiology of AD includes oxidative damage, which negatively impacts biological macromolecules. Triggers the apoptotic process, causing macromolecular destruction. Interestingly, rosavin attenuated Aβ-induced macromolecular damages, thereby preserving neuronal integrity. Furthermore, the activation of antioxidative defense enzymes by rosavin inhibited oxidative damage. The positive outcomes associated with rosavin were primarily attributed to its capacity to enhance acetylcholine-mediated effects. Finally, rosavin has the potential to alleviate Aβ-induced neurotoxicity and macromolecular damages, ultimately resulting in enhanced memorial and reasoning function in Wistar rats, offering promising prospects for the treatment of AD.