AIM： To investigate the role of reactive oxygen species （ROS） in ethanol-mediated cell death of polarized hepatic （WlF-B） cells.METHODS： In this work, WIF-B cultures were treated with pyrazole （inducer of cytoc...AIM： To investigate the role of reactive oxygen species （ROS） in ethanol-mediated cell death of polarized hepatic （WlF-B） cells.METHODS： In this work, WIF-B cultures were treated with pyrazole （inducer of cytochrome P4502E1, CYP2E1） and/or L-buthionine sulfoximine （BSO）, a known inhibitor of hepatic glutathione （GSH）, followed by evaluation of ROS production, antioxidant levels, and measures of cell injury （apoptosis and necrosis）.RESULTS： The results revealed that ethanol treatment alone caused a significant two-fold increase in the activation of caspase-3 as well as a similar doubling in ROS. When the activity of the CYP2E1 was increased by pyrazole pretreatment, an additional two-fold elevation in ROS was detected. However, the CYP2EIrelated ROS elevation was not accompanied with a correlative increase in apoptotic cell injury, but rather was found to be associated with an increase in necrotic cell death. Interestingly, when the thiol status of the cells was manipulated using BSO, the ethanol-induced activation of caspase-3 was abrogated. Additionally, ethanol-treated cells displayed enhanced susceptibility to Fas-mediated apoptosis that was blocked by GSH depletion as a result of diminished caspase-8 activity.CONCLUSION： Apoptotic cell death induced as a consequence of ethanol metabolism is not completely dependent upon ROS status but is dependent on sustained GSH levels,展开更多
基金Supported by The National Institute on Alcohol Abuse and Alcoholism and by the Department of Veterans Affairs
文摘AIM： To investigate the role of reactive oxygen species （ROS） in ethanol-mediated cell death of polarized hepatic （WlF-B） cells.METHODS： In this work, WIF-B cultures were treated with pyrazole （inducer of cytochrome P4502E1, CYP2E1） and/or L-buthionine sulfoximine （BSO）, a known inhibitor of hepatic glutathione （GSH）, followed by evaluation of ROS production, antioxidant levels, and measures of cell injury （apoptosis and necrosis）.RESULTS： The results revealed that ethanol treatment alone caused a significant two-fold increase in the activation of caspase-3 as well as a similar doubling in ROS. When the activity of the CYP2E1 was increased by pyrazole pretreatment, an additional two-fold elevation in ROS was detected. However, the CYP2EIrelated ROS elevation was not accompanied with a correlative increase in apoptotic cell injury, but rather was found to be associated with an increase in necrotic cell death. Interestingly, when the thiol status of the cells was manipulated using BSO, the ethanol-induced activation of caspase-3 was abrogated. Additionally, ethanol-treated cells displayed enhanced susceptibility to Fas-mediated apoptosis that was blocked by GSH depletion as a result of diminished caspase-8 activity.CONCLUSION： Apoptotic cell death induced as a consequence of ethanol metabolism is not completely dependent upon ROS status but is dependent on sustained GSH levels,
基金Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry of China(No.2008-890)Sci-Tech Project of Higher Educational and Scientific Research Institutions of DongguanCity(No.2008108101035)~~
