Curcumin Ameliorates Palmitic Acid-Induced Saos-2 Cell Apoptosis Via Inhibiting Oxidative Stress and Autophagy

Figure 6

Curcumin inhibited oxidative stress and autophagy in H₂O₂- induced Saos-2 cells apoptosis. Saos-2 cells were incubated with H₂O₂ or AY-22989 for 24 h with or without curcumin. The generation of ROS was measured by using the DCFH-DA. Saos-2 cells were stained with DCFH-DA and analyzed by Fluorescent microscopy (a). Statistical analysis of fluorescence intensity in Saos-2 cells by image J software (b). The relative levels of SOD were detected by a microplate reader (c). Western blot detected ATG5, P62, LC3I and LCII (d). Ratio of ATG5 and P62 to GAPDH, and ratio of LC3- II to LC3- I (e) (f), and (g). Fluorescent microscopy analysis of Saos-2 cells transfected with mCherry-GFP-LC3B adenovirus (h). Statistical analysis of fluorescent dots in Saos-2 cells (i). Yellow spots indicate autophagosomes, and red spots indicate autolysosomes after the pictures were merged. Ctrl: control; Cur: 10 µM curcumin; H₂O₂: 100 µM H₂O₂; AY: 10 µM AY-22989. Statistical analysis is shown on the bar graphs. Data are presented as the mean ± SEM of the three independent experiments. versus control, versus PA.

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