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Cell-free and cell-based estrogenic assays both revealed that calycosin competitively bound with ERα and ERβ. In addition, calycosin also displayed selective potency and affinity to ERα and ERβ in reporter-gene assays. Clinical and animal studies have suggested multiple benefits of SERM, and several SERMs have already been clinically approved, including raloxifene and tamoxifen. Recent findings have demonstrated the beneficial effects of these two classical SERMs upon the vascular system , , , . Since raloxifene and tamoxifen share the same/similar antagonistic action with calycosin at ERβ, we compared the angiogenic effects of the three compounds in zebrafish embryos. Of the three, only calycosin promoted significant angiogenic development in the SIVs of zebrafish embryos.
A previous study investigated the activities of compounds demonstrated to be active in zebrafish embryo bioassays, in both zebrafish and mammalian cell lines . Interestingly, only half of the 14 compounds were shown to be active in both embryos and either one of the cell lines, revealing that they exerted direct action upon cells. In our results, calycosin not only promoted angiogenesis in zebrafish but also enhanced endothelial cells proliferation and tube formation in HUVECs in vitro, both of which are standard tests for angiogenesis. Although no study has been carried out to identify bioequivalent doses between cell cultures and zebrafish, our results suggest that calycosin, at least in part, exerts direct action upon endothelial cells. Thus, we can further investigate the mechanism of action of calycosin in cell culture. 1e1e36bf2d