Modulation of β-amyloid (Aβ)-induced neurotoxicity has emerged as a possible therapeutic approach to ameliorate the onset and progression of Alzheimer's disease. The present study aimed to evaluate the protective effect of Inonotus obliquus extracts (IOEs) on Aβ-induced neurotoxicity in cultured rat pheochromocytoma (PC12) cells and rats. Exposure of PC12 cells to IOE significantly elevated cell viability, decreased intracellular calcium levels, and attenuated Aβ-mediated cell apoptosis. In aging rats, IOE can decrease the production of amyloid precursor protein (APP) and the levels of Aβ plaques in hippocampus after IOE treatment in the brain by an action that is associated with a lowering of the of IL-1β, TNF-α levels. Our findings indicate that IOE has potential neuroprotective actions against Aβ-induced neurotoxicity, which may occur through modulation of calcium channels or downstream molecules involved in inflammation.