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Biospecific isolation and characterization of angiogenesis-promoting ingredients in Buyang Huanwu decoction using affinity chromatography on rat brain microvascular endothelial cells combined with solid-phase extraction, and HPLC-MS/MS
Talanta ( IF 6.1 ) Pub Date : 2017-11-15 , DOI: 10.1016/j.talanta.2017.11.018
Fengyun Liao , Yingjiao Meng , Huazhu Zheng , Dongmei He , Xiao Shen , Jianye Yu , Yinai Wu , Lisheng Wang

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.



中文翻译:

固相萃取-HPLC-MS / MS对大鼠脑微血管内皮细胞进行亲和层析,对补阳还五汤中促血管新生成分进行生物特异性分离和表征

据报道,补阳还五汤具有促进血管生成的作用,但其有效成分尚不清楚。在这项研究中,我们开发了一种筛选BHD中潜在促血管生成化合物的方法,该方法涉及使用活大鼠脑微血管内皮细胞(rBMEC)进行生物特异性分离,并使用固相萃取(SPE)和高效液相色谱-串联质谱进行表征光谱分析(HPLC-MS / MS)。六种化合物显示出与rBMEC的结合亲和力,并进一步被鉴定为6-羟基山fer酚-二-O-葡萄糖苷,pa药苷,calycosin-7-O-β-D-葡萄糖苷,没药苷(galoylpaeoniflorin),甲霉菌素-7-O-β-D-葡萄糖苷, (3R)-7,2′-羟基-3′,4′-二甲氧基-异黄烷。结果表明,除6-羟基山emp酚-二-O-葡萄糖苷外,其中五种对rBMECs具有抑制氧葡萄糖剥夺/再灌注损伤的作用,并上调了血管内皮生长因子和碱性成纤维细胞的分泌,提示其潜在机制血管生成活性。我们的研究结果表明,基于生物特异性活细胞的分离结合SPE和HPLC-MS / MS是筛选中药中潜在生物活性成分的有效方法。

更新日期:2017-11-15
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