Biatractylolide is a Chinese medicinal compound used to treat various diseases. However, the molecular mechanisms of biatractylolide in preventing and protecting Alzheimer’s disease (AD) remained elusive. This study was conducted to observe the effect of biatractylolide on the pathological changes of amyloid beta protein (Aβ)-induced AD. In vitro assays (MTT and flow cytometry) were applied to detect the effect of biatractylolide on PC12 cell proliferation, growth inhibition rate and apoptosis. In order to assess the spatial learning and memory abilities of AD rats, Morris water maze model was applied in vivo, and the activity of the NF-κB signaling pathway and concentrations of TNF-α, IL-6, and IL-1β were measured. The results demonstrated that biatractylolide can reduce apoptosis in hippocampal cells, prevent and improve the cognitive decline induced by Aβ, prevent the morphological changes in hippocampal nerve cells and can reduce the activation of NF-κB signal pathway in vivo. Taken together, biatractylolide appeared to be a useful agent for the treatment of Aβ-related pathologies in the central nervous system, and can be considered for therapeutic application in AD-affected patients.