Identification of mundoserone by zebrafish in vivo screening as a natural product with anti-angiogenic activity

The present study aimed to screen natural products with anti-angiogenic potential from the Natural Products Collection of MicroSource. The anti-angiogenic activity of 240 natural products was assessed using the zebrafish line Tg(fli1a: EGFP) . At 24 h post-fertilization, the embryos were treated with the library compounds for 24 h and, the morphology of the intersegmental vessels (ISVs) was then assessed using a fluorescence microscope, followed by counting of ISVs and calculation of the inhibition ratio. The expression of angiogenesis-associated genes was determined by quantitative polymerase chain reaction. The results indicated that mundoserone inhibited ISV formation in zebrafish embryos in a dose-dependent manner, with a significant anti-angiogenic activity observed at a concentration of 10 µM, leading to an ISV inhibition ratio of 73.6±1.3%. Mundoserone significantly reduced the expression of slit guidance ligand 3 (SLIT3), roundabout guidance receptor 1 (ROBO1) and -2, fibroblast growth factor receptor (FGFR)2 and -3, as well as protein tyrosine phosphatase, receptor type B (PTP-RB), but increased the expression of NOTCH1A. Accordingly, mundoserone may be an effective angiogenic inhibitor, which acts via downregulation of SLIT/ROBO1 and FGFR/PTP-RB, and upregulation of NOTCH1A signaling.