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

Figure 4

Curcumin reduces oxidative stress and oxidative stress-induced autophagy in PA-induced Saos-2 cell. Saos-2 cells were incubated with 200 μM PA for 24 h with or without curcumin, NAC, or 3-MA. 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 LC3II (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; PA, 200 mM palmitic acid; NAC, 2 mM N-acetylcysteine; 3-MA/3-MA, 5 mM 3-methyladenine. 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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