3.6.3. Pro-Angiogenesis Assay

Vascular insufficiency in zebrafish was modeled by VEGFR tyrosine kinase inhibitor PTK787 to evaluate the effects of compounds on pro-angiogenesis according to previous report [8,26]. The healthy zebrafish larvae were separated into 24-well plates (ten embryos per well) in a 2 mL final volume of culture water at 24 h post fertilization (hpf). 0.2 μg/mL PTK787 was co-treated with each test compound (30, 70, 120 μg/mL) as test group. The control group was fresh culture water, the model group was 0.2 μg/mL PTK787, the positive drug group was 0.2 μg/mL PTK787 and 120 μg/mL ginsenoside Rg1. After 24 h incubation in a light-operated incubator at 28.0 ◦C ± 0.5 ◦C, the number of intersegmental blood vessels (ISVs) were captured by a fluorescent microscope (Olympus, SZX2-ILLTQ, Tokyo, Japan). Intact and defective vessels were counted separately and ISVs index was defined as follows: ISV index = number of intact vessels × 1 + number of defective vessels × 0.5 [27]. The zebrafish larvae without PTK787 in test group was used to evaluate the effects of compounds on anti-angiogenesis under the same conditions above described. All treatments were performed in triplicate.
