Figure 3.

Acetylshikonin attenuated H₂O₂-induced ROS generation and mitochondrial membrane potential loss in both SH-SY5Y and PC12 cells with dose-dependent manner. The cells were treated with desired concentration of acetylshikonin for 12 hours and then stimulated with H₂O₂ (500 μM) for 2 hours, and the ROS generation and mitochondrial membrane potential loss were detected by H₂DCF-DA and Rhodamine 123 staining, respectively. Representative photograph of ROS generation in SH-SY5Y (a) or PC12 (b) cells was taken by fluorescence microscope; the quantitative analysis of ROS generation in SH-SY5Y (c) or PC12 (d) cells was measured by fluorescence spectrophotometer. Representative photograph of mitochondrial membrane potential loss in SH-SY5Y (e) or PC12 (f) cells was taken by fluorescence microscope; the quantitative analysis of mitochondrial membrane potential loss in SH-SY5Y (g) or PC12 (h) cells was measured by fluorescence spectrophotometer. Data shown are means ± SEM of results from independent experiments in triplicate. *P < 0.05 compared with control cells; ^# P < 0.05 compared with H₂O₂-stimulated cells.