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Diosbulbin B (DIOB) is the most abundant component of Dioscorea bulbifera L.(DB), a traditional Chinese medicine herb.DIOB and DB extracts have been reported to cause liver injury in animals.The mechanisms of DIOB-induced hepatotoxicity remain unknown.The objective of the present study was to investigate the role of metabolic activation in DIOB-induced hepatotoxicity.DIOB was incubated with mouse, rat, and human liver microsomes, and a cis-enedione-derived metabolite was detected in the microsomal mixtures.The formation of the electrophilic metabolite was NADPH-dependent.The presence of ketoconazole decreased the production of the enedione.P450 3A4 was found to be responsible for the bioactivation of DIOB.