Figures index : In vitro Anti-angiogenic Effects of Tea Saponin and Tea Aglucone on Human Umbilical Vein Endothelial Cells : Science and Education Publishing

Figures index

Journal of Food and Nutrition Research. 2015, 3(3), 206-212 doi:10.12691/jfnr-3-3-13

Figure 2. Effect of TS on HUVEC morphology (A) and VEGF-induced proliferation (B). (A) After treated with varying concentrations of TS for 24 h, cell morphology was observed under inverted light microscopy. (B) VEGF-induced HUVEC proliferation was obtained by treatment with 5 μg/ml VEGF-A. After treated with varying concentrations of TS or TA for 24 h with presence of 5 μg/ml VEGF-A, cell viability was quantified using CCK-8 kit. Scale bar, 10 μm

Figure 3. Graphical representation of cell cycle analysis of HUVECs. After treated with varying concentrations of TS for 24 h, cell cycle analysis was performed using PI staining and flow cytometry. The cell population is expressed as percentage of the total cells analyzed. One representative experiment of three is shown

Figure 4. Flow cytometric evaluation of HUVEC apoptosis. Histograms derived from flow cytometry comparing apoptotic cells between vehicle and indicated concentrations of TS treated cells. After treatment for 24 h, the induction of apoptosis was determined using flow cytometric analysis of Annexin V-FITC and PI-stained HUVECs. Cells in the lower right quadrant indicate Annexin-positive, early apoptotic cells, and cells in the upper right quadrant indicate Annexin-positive/PI positive, late apoptotic cells. Data were the mean result from three parallel experiments

Figure 5. Transmission electron micrographs of HUVECs treated with vehicle (A) or 3 μM (B–E) of TS for 24 h. Control cells treated with vehicle exhibited a normal undifferentiated phenotype, showing absence of apoptotic bodies and nuclear condensation, whereas in the TS-treated cells, various kinds of autophagic vesicles (white arrowheads in B) are observed within the cytoplasm. These vesicles include the typical autophagosomes (Au in C), early autolysosomes (D) in which lysosomes (Ly) are still present, and late autolysosomes (E) with degraded amorphous substances. Scale bars represent 5 μm in A and B and 0.5 μm in C–E

Figure 6. Effect of TS (A) and TA (B) on HUVEC migration in wound healing assay. Monolayer HUVECs were wounded by scratching with pipette tips and treated with ethanol vehicle or different concentrations of TS or TA in serum-free EGM-2 medium. After 16 h of incubation, the migrated cells were quantified by manual counting. The percentage of inhibition was expressed using vehicle treated cells at 100%. The results are expressed as mean±SD (n = 5). Columns, mean; bars, SD. **, P<0.01; ***, P<0.001 versus vehicle-treated group. Scale bar, 10 μm

Figure 7. Effect of TS (A) and TA (B) on HUVEC invasion in Transwell assay. A total of 2 ×10⁴ HUVECs were seeded in the top chamber and treated with ethanol vehicle or varying concentrations of TS or TA. After 24 h, HUVECs that invaded through the membrane were stained with crystal violet and quantified. The percentage of inhibition was expressed using vehicle treated cells at 100%. The results are expressed as mean ± SD (n = 5). Columns, mean; bars, SD. *, P<0.05; ***, P<0.001 versus vehicle-treated group. Scale bar, 10 μm

Figure 8. Effect of TS (A) and TA (B) on HUVEC tube formation. After treated with vehicle or varying concentrations of TS or TA for 6 h, tubular structure in each group was measured using Image-pro Plus 6.0 System. The percentage of inhibition was expressed using vehicle treated cells at 100%. The results are expressed as mean ± SD (n = 3). Columns, mean; bars, SD. *, P<0.05; **, P<0.01, ***, P<0.001 versus vehicle-treated group. Scale bar, 10 μm