- Title
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Marine Cyclotripeptide X-13 Promotes Angiogenesis in Zebrafish and Human Endothelial Cells via PI3K/Akt/eNOS Signaling Pathways
- Authors
- Lu, X.L., Xu, Z.L., Yao, X.L., Su, F.J., Ye, C.H., Li, J., Lin, Y.C., Wang, G.L., Zeng, J.S., Huang, R.X., Ou, J.S., Sun, H.S., Wang, L.P., Pang, J.Y., and Pei, Z.
- Source
- Full text @ Mar. Drugs
Effects of X-13 on angiogenesis in TG (Fli-1:EGFP) zebrafish embryos. (A) Representative brightfield images of zebrafish larvae from 48 to 96 hpf. Treatment with X-13 for up to 96 h did not adversely affect the normal development of zebrafish larvae; (B) Representative fluorescence microscopy images; and (C) The bar chart shows quantitative data. X-13 (10, 50, 100 μM) dose-dependently induced angiogenesis in zebrafish embryos with EGFP expressing SIV. The angiogenic activity of X-13 at 50 μM and beyond was comparable to that of VEGF at 100 ng/mL. Data are expressed as the mean ± SD. Results were obtained from six independent experiments (* p < 0.05 vs. control vehicle group). |
The effects of X-13 on Human Umbilical Vein Endothelial Cell (HUVEC) invasion, migration and tube formation. HUVEC cultures were incubated with X-13 in the presence or absence of inhibitors for different periods in different assays and VEGF-treated cell cultures served as a positive control. Representative images show that X-13 induced HUVEC migration (A), invasion (B) and tube formation (C) in HUVEC cultures. L-NAME and LY294002 significantly inhibited X-13-induced cell invasion (A), migration (B) and tube formation (C). The bar chart shows quantitative data. Data are expressed as the mean ± SD. Results were obtained from four independent experiments (* X-13 vs. vehicle control group, p < 0.05; # LY294002 plus X-13 vs. X-13, p < 0.05; ☆ L-NAME plus X-13 vs. X-13, p < 0.05). |
Activation of Akt/PI3K pathways and nitric oxide (NO) production in X-13-treated HUVECs. Representative blots show that X-13 increased Akt and eNOS phosphorylation (A,B) while PI3K inhibitor LY294002 attenuated X-13-induced Akt and eNOS phosphorylation (C). Representative image shows that X-13 induced NO release while PI3K inhibitor LY294002, NOS inhibitor LAME, but not erk1/2 inhibitor PD98059 reduced X-13-induced NO release (D). The bar charts show quantitative data. Data are expressed as the mean ± SD. Results were obtained from three independent experiments (* X-13 vs. vehicle control group, p < 0.05; # LY294002 plus X-13 vs. X-13, p < 0.05; ☆ L-NAME plus X-13 vs. X-13, p < 0.05). |