calycosin-7-o-beta-d-glucopyranoside and galloylpaeoniflorin

Buyang Huanwu decoction (BHD) was reported to exert angiogenesis-promoting effects, but its active ingredients remain unknown. In this study, we developed a method to screen potential angiogenesis-promoting compounds in BHD, which involved biospecific isolation using live rat brain microvascular endothelial cells (rBMECs) and characterization using solid-phase extraction (SPE) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Six compounds showed binding affinity to rBMECs and were further identified as 6-hydroxykaempferol-di-O-glucoside, paeoniflorin, calycosin-7-O-β-D-glucoside, galloylpaeoniflorin, formononetin-7-O-β-D-glucoside, and (3R)-7,2'-hydroxy-3',4'-dimethoxy-isoflavan. The results indicated that five of them except 6-hydroxykaempferol-di-O-glucoside showed a protective effect against oxygen glucose deprivation/reperfusion injury in rBMECs and upregulated the secretion of vascular endothelial growth factor and basic fibroblast growth factor, suggesting a mechanism underlying their angiogenic activity. Our findings suggest that biospecific live cell-based isolation combined with SPE and HPLC-MS/MS is an effective method for screening potential bioactive components in traditional Chinese medicines.

Topics: Angiogenesis Inducing Agents; Animals; Animals, Newborn; Brain; Bridged Bicyclo Compounds, Heterocyclic; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Endothelial Cells; Glucosides; Isoflavones; Monoterpenes; Primary Cell Culture; Rats; Rats, Sprague-Dawley; Solid Phase Extraction; Tandem Mass Spectrometry