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首页> 外文期刊>Journal of separation science. >Screening vasoconstriction inhibitors from traditional Chinese medicines using a vascular smooth muscle/cell membrane chromatography-offline-liquid chromatography-mass spectrometry
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Screening vasoconstriction inhibitors from traditional Chinese medicines using a vascular smooth muscle/cell membrane chromatography-offline-liquid chromatography-mass spectrometry

机译：血管平滑肌/细胞膜色谱-离线-液相色谱-质谱法从中药中筛选血管收缩抑制剂

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摘要

We developed an analytical method for screening vasoconstriction inhibitors from traditional Chinese medicines (TCMs) by combining vascular smooth muscle/cell membrane chromatography (VSM/CMC) with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Primary cultured VSM cells from rat thoracic aortas were used for preparation of the stationary phase of the VSM/CMC column. Retention fractions from the VSM/CMC column were collected and then analyzed by LC-MS/MS under the optimized conditions offline. The suitability and reliability of the VSM/CMC-offline-LC-MS/MS method was assessed using nitrendipine and nifedipine as positive controls, and this method was then applied to screen vasodilator components from the extracts of Fructus Schisandrae Chinensis (FSC) and Fructus Schisandrae Sphenantherae (FSS). The major components from both species retained by VSM/CMC were identified as deoxyschizandrin (DSD) and schisantherin A (STA) by LC-MS/MS. Competition experiments indicated that DSD and nifedipine bound competitively to membrane receptors, while DSD and STA had partly overlapping binding sites on VSM-cell membranes. In vitro pharmacological trials confirmed that STA and DSD could dose-dependently relax the rat thoracic aortas pre-contracted by KCl. Our VSM/CMC-offline-LC-MS/MS method can be applied for screening vasoconstriction inhibitors from TCMs collected from FSC and FSS, and may be useful in the development of vasodilators from natural products.
机译：通过将血管平滑肌/细胞膜色谱法（VSM / CMC）与液相色谱-串联质谱法（LC-MS / MS）相结合，我们开发了一种从中药（TCM）中筛选血管收缩抑制剂的分析方法。将来自大鼠胸主动脉的原代培养的VSM细胞用于制备VSM / CMC色谱柱的固定相。收集来自VSM / CMC色谱柱的保留级分，然后在最佳条件下离线通过LC-MS / MS分析。以硝苯地平和硝苯地平为阳性对照评估了VSM / CMC-offline-LC-MS / MS方法的适用性和可靠性，然后将该方法用于从五味子和五味子提取物中筛选血管扩张剂成分。五味子五味子（FSS）。 VSM / CMC保留的两种物种的主要成分通过LC-MS / MS鉴定为脱氧五味子素（DSD）和五味子素A（STA）。竞争实验表明，DSD和硝苯地平与膜受体竞争性结合，而DSD和STA在VSM细胞膜上具有部分重叠的结合位点。体外药理试验证实，STA和DSD可以剂量依赖性地松弛由KCl预收缩的大鼠胸主动脉。我们的VSM / CMC-offline-LC-MS / MS方法可用于从FSC和FSS收集的中药中筛选血管收缩抑制剂，并可用于开发天然产物的血管扩张剂。

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