Dosage effects of resveratrol on ethanol-induced cell death in the human K562 cell line.
Previous studies have established that ethanol induces cell apoptosis and necrosis. However, the precise molecular mechanisms are currently unclear. Here, we show that higher concentrations of ethanol (250-400 mM) induced a shift from apoptotic to necrotic cell death in human K562 cells, and that resveratrol, a grape-derived phytoalexin with known antioxidant and anti-inflammatory properties, inhibited or enhanced ethanol-induced apoptosis/necrosis depending on the treatment dosage. Using the cell permeable dye 2',7'-dichlorofluorescin diacetate (DCF-DA) as an indicator of reactive oxygen species (ROS) generation, we showed that ethanol treatment directly increased intracellular oxidative stress. This intracellular oxidative stress increased in response to high concentrations (100-200 microM) of resveratrol, but remained unchanged following treatment with low concentrations (10-25 microM) of resveratrol. Further studies showed that resveratrol could attenuate or enhance ethanol-induced intracellular oxidative stress generation-dependent on treatment dosage, and that this effect could be correlated with cell apoptosis or necrosis. Importantly, ethanol-induced changes in intracellular ATP levels were also correlated with resveratrol dosage. Taken together, these results indicate that the treatment dosage may determine the effect of resveratrol on ethanol-induced ROS generation, intracellular ATP levels, and cell apoptosis or necrosis. Thus our findings support the possibility that appropriate dosage of resveratrol aids in decreasing the toxic effect of ethanol.
Chan
Wen-Hsiung
WH
Department of Bioscience Technology and Center for Nanotechnology, Chung Yuan Christian University, 200 Chung Pai Road, 32023 Chung Li, Taiwan. whchan@cycu.edu.tw
Chang
Ying-Jing
YJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004789:Enzyme Activation / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D006801:Humans; D020014:K562 Cells; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D048670:MAP Kinase Kinase 4 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*; D013997:Time Factors
D017382:Reactive Oxygen Species; D013267:Stilbenes; D000431:Ethanol; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D048670:MAP Kinase Kinase 4; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D000077185:Resveratrol