Figure 4.
Effects of 3,5,7-trimethoxyflavone (6) on intracellular ROS accumulation in TNF-α stimulated normal human dermal fibroblasts (NHDFs). (A) Bands of protein expressions. (B) Bar graphs of the relative expression. We seeded NHDFs at 1 × 104 cells/well in 96-well black plates incubated them for 24 h, and replaced the medium with a serum-free medium to create starvation conditions. After 24 h, cells were treated with 50 and 100 μM of 6 for 1h, and the cells were exposed to 20 ng/mL TNF-α for 15 min. The cell was stained with dichlorofluorescin diacetate (DCFDA) for 15 min, and photographs was observed with a microscope IX51. The measurement of fluorescence was conducted using a SPARK 10M. Results of intracellular ROS were presented as a percent of the vehicle control. The results were obtained through three replicate experiments, and the graphs are represented as mean ± SEM. # p < 0.05 vs. vehicle control. * p < 0.05 and ** p < 0.01 vs. TNF-α stimulated control.