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

Figure 4

Quercetin decreased intralysosomal redox-active iron and it induced ROS in lysosome as well as DFO. The mice primary hepatocytes were isolated according to a two-step collagenase method and treatment for 6 hours. The intralysosomal iron was measured by SSM ((a) the exposure time was 50 min in the dark, ) and quantitated by flame atomic absorption spectrophotometry (b). The cells were incubated with a combination of LysoTracker (to detect lysosomes) and H₂DCFDA (to detect ROS production) for 30 min and immediately observed by confocal microscopy with sequential recording in green (oxidized product of H₂DCF) and red (LysoTracker). All photomicrographs were taken at ×200 (c, ). C: control; E: ethanol; EQ: ethanol plus quercetin; ED: ethanol plus DFO; Q: quercetin; D: DFO. A: versus the control; B: versus the ethanol group.