Journal
TALANTA
Volume 179, Issue -, Pages 490-500Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2017.11.018
Keywords
Buyang Huanwu decoction; Solid-phase extraction; Biospecific live cell-based isolation; Angiogenesis; Oxygen-glucose deprivation and reperfusion; Traditional Chinese medicines
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Funding
- National Natural Science Foundation of China [81373973]
- Natural Science Foundation of China [81573872]
- Guangdong Provincial Science and Technology projects [2013B032500015]
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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-beta-D-glucoside, galloylpaeoniflorin, formononetin-7-O-beta-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.
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