Figures index

From

Artepillin C Suppresses Angiogenesis by Inhibiting Tube-Formation and Inducing Apoptosis of Endothelial Cells

Mok-Ryeon Ahn, Sun Wook Kim, Shigenori Kumazawa, Toshiro Ohta

Journal of Food and Nutrition Research. 2013, 1(5), 92-96 doi:10.12691/jfnr-1-5-3
  • Figure 1. ARC inhibits tube formation and induces reduction of angiogenesis-related adhesion molecules in tube-forming HUVECs. A:HUVECs were sandwiched between two layers of collagen gel and induced to form blood vessel-like tubes. The cells were treated with various concentrations of ARC (3.13, 12.5, and 50 µg/mL). Tube-forming HUVECs in a 2-D culture model were observed at 6, 12, and 24 h. Each experiment was repeated at least three times and representative data are shown. Bar indicates 100 μm. B: ARC blocked the tube formation of HUVECs in a 2-D culture model. The tube-forming cells were treated with various concentrations of ARC for 24 h and then fixed. The fixed cells were stained with 0.1% toluidine blue. (a) 0 µg/mL, (b) 3.13 µg/mL, (c) 12.5 µg/mL, (d) 50 µg/mL. Representative photos are shown. Bar indicates 200 μm. Values are expressed as means ± SE (n = 3). *P < 0.05 vs. the control group.C: Cellular proteins were collected from tube-forming HUVECs that were treated with various concentrations of ARC(3.13, 12.5, and 50 µg/mL) for 12 and 36 h. Changes in VE-cadherin and PECAM-1 were analyzed by Western blotting. Each experiment was repeated at least three times and representative data are shown
  • Figure 2. ARC induces apoptosis in tube-forming HUVECs. A: The tube-forming cells were treated with various concentrations of ARC (3.13, 12.5, and 50 µg/mL) for 24 h, fixed and stained with DAPI to observe cell nuclei. (a) 0 µg/mL, (b) 3.13 µg/mL, (c) 12.5 µg/mL, (d) 50 µg/mL. Representative photos are shown. Bar indicates 50 μm. B: Rates of apoptosis (percentage of condensed and fragmented cell nuclei against total cell nuclei) were quantified. A total of more than 500 cells from six fields were counted for each data point. Values are expressed as means ± SE (n = 6). *P < 0.05, **P < 0.01vs. the control group
  • Figure 3.ARC inactivates ERK 1/2 and induces activation of caspase cascade. A, B: Cellular proteins were collected from tube-forming HUVECs that were treated with various concentrations of ARC(3.13, 12.5, and 50 µg/mL) for 6, 12, and 24 h. (A) Changes in phosphorylation state of ERK 1/2, p38, and Akt and (B) caspase-3, PARP, andlamin A/Cwere analyzed by Western blotting. Each experiment was repeated at least three times and representative data are shown