Relationship between oxidative stress and hepatic glutathione levels in ethanol-mediated apoptosis of polarized hepatic cells

Figure 1 Ethanol-induced protein oxidation, oxidative stress, and caspase activation in WIF-B cells. WIF-B cultures were cultured for 48 h in the absence or presence of ethanol, and DAS or NAC when indicated. As a positive control for H₂O₂ production, some cultures were treated with 2 &mgr;mol/L Antimycin A (C + AA) for 30 min prior to analysis. A: Protein oxidation analysis represented as DNP-specific proteins detected in WIF-B cell lysates using the OxyBlot protein oxidation analysis; B: H₂O₂ production was detected by dichlorofluorescein fluorescence quantification. Data from 5 independent experiments is represented as the amount of fluorescent (DCFDA) oxidized products detected in the treated cells per mg protein; C: The activation of the proapoptotic protease, caspase-3, was detected as described in “Material and Methods” for five independent experiments. ^aP < 0.05 vs control; ^cP < 0.05 vs ethanol alone-treated cells.