Iron-Mediated Lysosomal Membrane Permeabilization in Ethanol-Induced Hepatic Oxidative Damage and Apoptosis: Protective Effects of Quercetin

Figure 6

The role of lysosomal iron in ethanol-induced hepatocytes damage and the protective effect of quercetin. Mouse primary hepatocytes grown on coverslips were treated with ethanol (100 mmol/L, 24 hours), bafilomycin A1 (100 nmol/L, 4 hours), FeCl₃ (60 μmol/L, 4 hours), DFO (1 mmol/L, 4 hours), and quercetin (100 μmol/L, 24 hours). The intralysosomal redox-active iron content (a) and lysosome status (b) were measured by SSM (the exposure time for the FeCl₃ group was 20 min; other groups were 50 min) and AO-uptake technique, respectively. All photomicrographs were taken at ×200 (). The leakage of AST and LDH from hepatocytes (c, d, ), together with cellular ROS production (e, ), was detected by spectrophotometry. MMP (f and g) and apoptosis (h) were measured by flow cytometry (). Each value represents the mean ± SD. C: control; E: ethanol; EI: ethanol plus FeCl₃; EB ethanol plus bafilomycin A1; ED: ethanol plus DFO. A: versus the control; B: versus the ethanol group.