Fig. 3
Effect of C3G on glutamate-induced ROS generation in HT22 cells and antioxidant activity. (a) The pretreatment of cells with C3G (0-1 μM) for 24 h, followed by 5 mM glutamate for 18 h. After that the cells were incubated with 10 μM DCFH2-DA for 45 min at 37 °C. The fluorescence intensity was measured by flow cytometry. (b) Data were expressed as a relative ROS level of non-treated control. (c) The free radical scavenging activity of various concentrations of C3G (0.05–1 μM) was evaluated using DPPH assay. (d) Cells were pretreated specific ROS inhibitors for 30 min and followed by 5 mM glutamate for 18 h, cell viability was detected by the MTT assay. Glu, glutamate; NAC, N-acetyl-L-cysteine; GSH, glutathione; CAT, catalase; MnTRAP, Mn (III) tetrakis (4-benzoic acid) porphyrin; DM, deferoxamine mesylate. Values are the mean ± SD (n = 4). #p < 0.05 versus non-treated control, *p < 0.05, **p < 0.01, ***p < 0.001 versus 5 mM glutamate-treated cells