“Unraveling the Neuroprotective Mechanism of Dioscorea bulbifera (DB) in Alzheimer’s Disease (AD) Caused by Estrogen Loss: An Integrated Network Pharmacology Investigation and Experimental Validation”

Abstract Background Alzheimer’s disease (AD) is a leading cause of dementia, characterized by cognitive decline, and is more prevalent in women, possibly due to estrogen loss after menopause. Dioscorea bulbifera (DB), a medicinal plant, has been proposed as a potential treatment for AD. However, the underlying mechanism of action of DB and its neuroprotective effects in AD, particularly in the context of estrogen loss, remain unclear. Method In this study, we employed an integrative network pharmacology approach to predict the mechanism of action of DB in AD. Using a collection of AD‐related genes and predicted DB targets, we identified putative targets, direct regulatory targets, and potential regulatory targets of DB. Pathway‐enrichment analysis was performed to elucidate the pivotal pathways involved in DB’s treatment of AD. Molecular docking was conducted to verify the interactions between the core targets and the active ingredients of DB. In vivo experiments were conducted using ovariectomized rats induced with scopolamine to evaluate the neuroprotective effects of DB. Acetylcholine (Ach) and serum estradiol levels were quantified using ELISA. Result Our results identified 132 putative targets, including 68 direct regulatory targets and 25 potential regulatory targets of DB for the treatment of AD. Pathway‐enrichment analysis revealed that neurotransmitter clearance in the synaptic cleft was a crucial pathway in the treatment of AD with DB. Molecular docking further supported the interactions between the core targets (ESR1, APP, GSK3β, BACE1, AChE, and MAOB) and the active ingredients of DB. In vivo experiments using ovariectomized rats induced with scopolamine demonstrated the neuroprotective effect of DB and validated the predicted mechanism of action. Conclusion Our findings provide experimental support for the predicted mechanism of action of DB in AD caused by estrogen loss, and validate its neuroprotective effects using behavioral tests and ELISA in ovariectomized rats. DB may hold promise as a potential therapeutic option for AD, particularly in the context of estrogen loss, and further research is warranted to explore its full potential in AD treatment.