SOD2 Mediates Amifostine-Induced Protection against Glutamate in PC12 Cells

Figure 6

SOD2-siRNA abolished amifostine-induced effects on mitochondrial superoxide production and intracellular ROS, GSH, and CAT levels. The PC12 cells were divided into six groups: control: cells cultured in drug-free medium; Ami: cells exposed to 500 μM amifostine for 24 h; Glu: cells treated with 15 mM glutamate for 24 h; Ami + Glu: cells exposed to 500 μM amifostine plus 15 mM glutamate for 24 h; SOD2-siRNA + Ami + Glu: cells incubated with SOD2-siRNA transfection complexes for 24 h and then exposed to 500 μM amifostine plus 15 mM glutamate for 24 h; SC-siRNA + Ami + Glu: cells incubated with scrambled- (SC-) siRNA transfection complexes for 24 h and then exposed to 500 μM amifostine plus 15 mM glutamate for 24 h. (a) Mitochondrial superoxide production (red) was measured by MitoSOX Red staining followed by image pro-plus software analysis, and nuclei were counterstained with DAPI (blue). Intracellular ROS (b), GSH (c), and CAT (d) levels were evaluated by the corresponding reagent kits, and the photographs of ROS fluorescence staining were recorded by a confocal microscope. Results are means ± SD, , NS: no significance, and bar = 10 μm.

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