AA-PMe and AA treatment results in gross defects in zebrafish vascular development. (A) Live fluorescence microscopy images depicting gross morphology of 72 hpf Tg(fli:EGFP) zebrafish embryos treated with DMSO alone or the indicated concentrations of AA-PMe and AA. ISV and SIV are clearly defined by fluorescence, the latter appearing as a smooth basket-like structure with five to six arcades. Red and yellow bars indicate the lumina of the DA and PCV, respectively. Scale bar, 50 μm. (B) Length of the ISV; (C) the number of SIV branch points; (D) SIV length in 72 hpf embryos treated with DMSO or the indicated concentrations of AA-PMe and AA were determined as described. (E) DA lumina diameters. (F) PCV lumen diameter. Data are expressed as mean ± SEM from three independent experiments. *P<0.05, **P<0.01.
Abbreviations: AA, asiatic acid; AA-PMe, N-(2α,3β,23-acetoxyurs-12-en-28-oyl)-L-proline methyl ester; HUVEC, human umbilical vein endothelial cell; DMSO, dimethyl sulfoxide; ISV, intersegmental blood vessel; SIV, subintestinal vessel; DA, dorsal aorta; PCV, posterior cardinal vein.