FIGURE SUMMARY
Title

Kaempferol Identified by Zebrafish Assay and Fine Fractionations Strategy from Dysosma versipellis Inhibits Angiogenesis through VEGF and FGF Pathways

Authors
Liang, F., Han, Y., Gao, H., Xin, S., Chen, S., Wang, N., Qin, W., Zhong, H., Lin, S., Yao, X., Li, S.
Source
Full text @ Sci. Rep.

Fraction DYVE-D3 and kaempferol inhibited the outgrowth of ISVs.

Embryos at 48 hpf anterior is to left and dorsal is to top. Right column is enlargement of left column. (A,B), control group, ISV fully developed. (C,D) DYVE group, embryonic trunk formation was impaired and the head was small, while the ISVs were still present. (E,F) DYVE-D group, the embryonic trunk was very short, and ISVs developed. (G,H) DYVE-D3 group, the embryo developed slower than the control, but did not have strong defects. Outgrowth of ISV was inhibited. (I,J) kaempferol group, embryos developed slower than control, but were mostly normal. Outgrowth of ISVs was inhibited. (K,L) DYVE-B group, the embryos were slightly abnormal on the morphology, but ISVs appeared normal. White arrows point to ISV. Phenotype ratio is showed in the lower right corner. The working concentration of DYVE, DYVE-D, DYVE-D3 and DYVE-B was based on Supplementary Table S1, and kaempferol group was 40 µM.

Kaempferol inhibits VEGF-induced migration and tube formation of HUVECs

(A) upper row, kaempferol inhibited VEGF-induced HUVECs migration in wound healing assay. HUVECs were allowed to grow into full confluence in 24–well plates. Cells were wounded with a pipette and treated with or without 40 ng/mL VEGF and different concentrations of kaempferol in ECM supplemented with 0.5% (v/v) FBS. Compared to control group, 40 µM kaempferol blocked induction by VEGF. Bottom row, kaempferol inhibited VEGF-induced HUVEC tube formation. (B,C) quantification of the results in (A). Image J was applied for statistical branching point numbers and the distance of remained gap. Scale bar, 100 µm. *P < 0.05 versus control group.

Kaempferol blocked PI3K/Akt and p38 MAPK in vitro and had a synergistic effect with PTK787/SU5402 on VEGF and FGF pathways in vivo.

(A) different concentrations of kaempferol were added to HUVECs for 24 hours or 48 hours and analysed for Erk1/2, p38, Akt phosphorylation and VEGFR2 expression by Western blotting. CTL-control. (B) a synergistic effect of kaempferol with VEGF inhibitor PTK787 or FGF inhibitor SU5402 was observed. Compounds were added at 6 hpf and pictured at 52 hpf, anterior is to left and dorsal is to top. Affected ratio is showed in the lower right corner. Full-length blots are presented in Supplementary Fig. S2.

Acknowledgments
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