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The water extract of Dancheonhwan (DCH) has been used to treat ischemic brain and heart damage in oriental medicine. However, little is known about the mechanism by which the water extract of DCH rescues cells from ischemic damage. Therefore, this study was designed to investigate the protective mechanisms of DCH on the H2O2-induced toxicity in H9c2 cardiomyoblast cells. Treatment of H2O2 markedly decreased the viability of H9c2 cardiomyoblast in a dose-dependent and time-dependent manner. The nature of H2O2-induced toxicity of H9c2 cells resulted from apoptotic death confirmed with genomic DNA fragmentation. DCH increased the viability of H2O2-treated H9c2 cells by about 23%, and partially suppressed the genomic DNA fragmentation and PARP cleavage. H2O2 also activated caspase-3 protease and -9 protease, but not both caspase-6 protease and -8 protease. H2O2 induced the mitochondria dysfunction, including mitochondria membrane permeability transition (MPT) and cytosolic release of cytochrome c from mitochondria, which was prevented in part by pretreatment of DCH. N-acetylcystein (NAC), a free-radical scavenger, alone increased the viability of H2O2-treated H9c2 cells in a dose-dependent manner. Furthermore, the combination of NAC with DCH significantly increased the viability of the H2O2-treated H9c2 cells in a dose-dependent manner. These data indicate that DCH has the protective effect on ROS-induced apoptosis of cadiomyoblast H9c2 cells.