Fig. 3

Alkaloid extract of YHS limited angiogenesis in vivo (a) Antiangiogenic effects of alkaloid extract of YHS in mice. C57BL/6 Mice were subcutaneously injected with 500 μl of a Matrigel mixture containing 50 ng VEGF. The mice treated with various concentrations of alkaloid extract of YHS (a: 0 mg/kg; b: 15 mg/kg; c: 30 mg/kg; d: 60 mg/kg) were euthanized 14 days later and Matrigel plugs were harvested. 4 μm paraffin sections were stained with CD31 (brown) and a representative image is shown, scale bar: 200 μm. b Quantification of relative vessel area (CD31 positive staining). Relative CD31 positive staining treated with 0 mg/kg alkaloid extract of YHS was set as 100%. Data are presented as mean ± SEM. ***, P < 0.001; *, P < 0.05, n = 4, paired t-test. c Representative images of microvessels formation on in vivo CAM model after the treatment of different concentrations of alkaloid extract of YHS (a: 0 μg/ml; b: 10 μg/ml; c: 50 μg/ml; d: 100 μg/ml). d Quantification of number of blood vessels in CAM model. The index was defined as the mean number of visible microvessel with the defined area of drug-containing pellets. Relative blood vessel number treated with 0 μg/ml alkaloid extract of YHS (control) was set as 100%. Data are presented as mean ± SEM. ***, P < 0.001; *, P < 0.05, n = 4, paired t-test