Purpose: Little is known regarding the therapeutic role of exercise against the risk of a high-fat diet (HFD) for Alzheimer's disease (AD) and AD-like cognitive deficits. This study aimed to investigate the therapeutic effect of treadmill running against HFD-induced progression in AD neuropathology and cognitive impairments in the tripletransgenic AD (3xTg-AD) mice. Methods: The 3xTg-AD mice were assigned to a chow diet (control, n = 10), an HFD (n = 10), or an HFD combined with exercise (HFD + EX, n = 10) group. Mice in the HFD were fed with a 60% fat diet for 20 wk. The HFD + EX mice were additionally subjected to treadmill running. Results: Compared with the control mice, the HFD mice had impaired brain insulin signaling, exacerbated AD neuropathology, defects in synaptic stability/plasticity, and apoptotic neuronal cell death in conjunction with exacerbated cognitive deficits in the affected brain regions, which were all significantly alleviated in the HFD + EX mice. Conclusion: The current findings suggest that treadmill running protects against AD-like disease progression and cognitive deficits caused by an HFD in the 3xTg-AD mice.