Ganoderma Triterpenoids Exert Antiatherogenic Effects in Mice by Alleviating Disturbed Flow-Induced Oxidative Stress and Inflammation

Figure 7

GTs ameliorated H₂O₂-induced oxidative stress in the ECs. (a) HUVECs were preincubated with GTs (500 μg/ml) or vehicle (Veh) control DMSO for 1 h before treated with or without H₂O₂ (400 μM, 30 min). Cells were loaded with the fluorescent dye DCFDA for measuring the levels of reactive oxygen species (ROS) using flow cytometry. The upper panel shows a representative histogram overlay of the ROS fluorescent intensities. Quantitative fluorescence intensities are means ± SEM of triplicate experiments. (b) Cells were preincubated with GTs (500 μg/ml) or Veh (DMSO) for 1 h before treated with or without H₂O₂ (400 μM) for 24 h. Cells were then stained with annexin V/PI and analyzed for apoptosis. The percentage of the apoptotic cells includes the numbers shown in the lower right Q3 (early apoptosis) and in the upper right Q2 (late apoptosis) quadrants. Data are means ± SEM of triplicate experiments. (c) HUVECs pretreated with GTs (500 μg/ml, 1 h) or Veh (DMSO) were treated with or without H₂O₂ (400 μM) for 5 h, and followed by immunostaining with anti-ET-1 (red) or anti-MCP-1 (red) antibodies as indicated, and counterstaining with DAPI. The fluorescence intensity was quantified using the NIH ImageJ program. Data are means ± SEM of triplicate experiments. Bar: 25 μm. Statistical significance was calculated using two-way ANOVA and post hoc Tukey’s tests, .

(a)

(b)

(c)