Alzheimer’s disease (AD) is a neurodegenerative disease afflicting many people worldwide. As the specific biomarker, beta amyloid (Aβ) is considered to serve a central role in the progress of AD. To discover effective therapy for AD, esculetin was investigated using SH-SY5Y cells to evaluate its protective effects against the neurotoxicity induced by Aβ25-35. As a result, esculetin can improve the viability of SH-SY5Y cells injured by Aβ25-35 (58.0%, 66.1% and 82.1% at 0.1, 1 and 10 μM vs 49.5% at 0 μM). Further investigation has demonstrated the protective effects of esculetin at different concetrations of 0.1, 1 and 10 μM are associated with its inhibition of oxidative stress and apoptosis in SH-SY5Y cells, which is more observable especially at both 1 and 10 μM. These observations can give evidences for the following investigation in vivo and discovery of novel preventive method for AD.