Anti-angiogenic effects of Qingdu granule on breast cancer through inhibiting NFAT signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Qingdu granule (QDG), a traditional Chinese herbal prescription, had anti-tumor effect on breast cancer. However the underlying mechanism of QDG was unclear. THE AIM OF THIS STUDY: The present study aimed to investigate whether QDG could inhibit angiogenesis of breast cancer via acting on nuclear factor of activated T-cells (NFAT) signaling pathway. This was implicated in human umbilical vein endothelial cells (HUVECs) in vitro and breast cancer xenograft model in vivo. MATERIALS AND METHODS: The VEGF165 (15.58 ng/mL) induced human umbilical vein endothelial cells (HUVECs) were treated with serum samples containing tamoxifen (TAM), tacrolimus (FK506), or QDG with three dosages. The migration and canalization capacities of HUVECs were evaluated by transwell migration and tube formation assay. In 72 h-cultured HUVECs, The gene expression, protein amount, and nuclear translocation of NFATc3 were measured. The anti-tumor and anti-angiogenic effects of QDG in vivo were investigated in breast cancer xenograft model. The serum VEGF levels, microvessel density, and protein expressions (immunohistochemistry and western blot) of VEGF, VEGFR2 and NFATc3 were detected. RESULTS: The results showed that, QDG significantly inhibited HUVEC migration and tube formation. It downregulated NFATc3 gene expression, decreased NFATc3 protein amount, and reduced the ratio of NFATc3 nuclear translocation in HUVECs. In breast cancer xenograft model, QDG treatment significantly suppressed tumor growth, inhibited VEGF release, and decreased microvessel density. QDG reduced protein expressions of VEGF, VEGFR2 and NFATc3. CONCLUSION: The results suggested that QDG showed anti-angiogenic effects of breast cancer both in vitro and in vivo. The mechanism might be partially associated with inhibiting NFAT signaling pathway.

Depletion of high-abundance flavonoids by metal complexation and identification of low-abundance flavonoids in Scutellaria baicalensis Georgi.

The complexation of metal cations and flavonoids with 5-hydroxyl or ortho-hydroxyl groups was successfully used for high-abundance flavone depletion from a botanical extract in this study. Due to their structural differences, five of the most highly abundant constituents, baicalin, wogonoside, baicalein, wogonin and oroxylin A, were successfully depleted from the ethanol extract of Radix Scutellariae. The depletion rates were approximately 99%, 85%, 99%, 70% and 76%, respectively. The recoveries of low-abundance constituents were very strong (approximately 70-100%). The efficiency of the low-abundance compounds' identification by high performance liquid chromatography electrospray tandem mass spectrometry (HPLC ESI MS/MS) was remarkable after the high-abundance constituents were removed. The number of compounds identified from the HPLC MS/MS data was 250% greater than the number of compounds identified in the untreated total extract. One hundred seventeen flavonoids were identified in the ethanol extract of Radix Scutellariae using this method, which was much greater than the number identified in previous studies without high-abundance constituent depletion. Among them, 13 sulphated flavonoids were identified. These low-abundance sulphated flavonoids can barely be detected in untreated total extracts. To the best of our knowledge, this is the first reported evidence that sulphated flavonoids have been identified from Radix Scutellariae. This method will facilitate the removal of high-abundance flavonoids and the identification of low-abundance compounds in botanical extracts.