Puerariaeflos (PF) is an oriental medical herb for alcohol abuse. To investigate whether PF possesses protective effects against ethanol (EtOH)-induced cytotoxicity in the central nervous system, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometric analysis, DNA fragmentation assay, and reverse transcription-polymerase chain reaction were performed on SK-N-MC human neuroblastoma cells. Cells treated with EtOH exhibited several apoptotic features, while those pre-treated with PF prior to EtOH exposure showed a decreased occurrence of apoptotic features. In addition, PF pre-treatment inhibited the EtOH-induced increase in caspase-3 mRNA expression. These results suggest that PF may exert protective effects against EtOH-induced apoptosis in human neuroblastoma cells.