Eupatilin (5,7-dihydroxy-3',4',6-trimethoxyflavone), an extract from Artemisia asiatica Nakai, is a flavonoid of pharmacologically active ingredients. Eupatilin is known to possess anti-cancer, anti-inflammatory, and anti-oxidative activity. Recently, eupatilin has been reported to be effective in producing gastric mucosal as an anti-gastritis agents. However, the mechanism of protective action is still unknown. We studied cytoprotective actions of eupatilin on H(2)O(2)-induced cell death and its possible mechanisms of action in human gastric (AGS) cells. Eupatilin dose-dependently inhibited H(2)O(2)-induced apoptosis as indicated by co-staining with Annexin V and propidium iodide. Hydrogen peroxide provoked phosphorylation of extracellular regulated kinase (ERK) and c-Jun NH(2)-terminal kinase (JNK), and activation of nuclear factor-kappaB (NF-kappaB). On the contrary, eupatilin decreased H(2)O(2)-induced activation of ERK, JNK and NF-kappaB. In addition, treatment of specific inhibitors for ERK, JNK, and NF-kappaB attenuated H(2)O(2)-induced apoptosis. Co-treatment of inhibitors and eupatilin was more effective in decreasing H(2)O(2)-induced apoptosis. Taken together, we suggest that eupatilin inhibits H(2)O(2)-induced apoptosis through the inhibition ERK, JNK, and NF-kappaB.