Comprehensive two-dimensional HepG2/cell membrane chromatography/monolithic column/time-of-flight mass spectrometry system for screening anti-tumor components from herbal medicines
Comprehensive two-dimensional HepG2/cell membrane chromatography/monolithic column/time-of-flight mass spectrometry system for screening anti-tumor components from herbal medicines
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DOI：
10.1016/j.chroma.2012.04.034
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发表时间：
2012-06-15
影响因子：
4.1
通讯作者：
Chai, Yifeng
中科院分区：
文献类型：
--
作者：
Chen, Xiaofei;Cao, Yan;Chai, Yifeng
细胞膜色谱是一种以特定细胞膜为固定相的生物亲和色谱方法。它已被证明是研究药物与膜受体之间结合相互作用的实用工具。本研究建立了一种新的全二维色谱法用于中药抗肿瘤活性成分的筛选。HepG 2/CMC模型首次被开发和应用作为第一维柱。采用带有两个进样环的自动十端口切换阀，将第一维组分引入到由整体柱和高分辨飞行时间质谱（TOFMS）组成的第二维组分中。基于HepG 2/CMC/整体柱/TOFMS系统的稳定性、选择性和适用性试验，筛选并鉴定了黄柏中的小檗碱（BBR）和四氢巴马汀（THP）、苦参中的氧化苦参碱和苦参碱为潜在活性成分。竞争置换实验表明，四种组分均能以与吉非替尼相似的方式作用于HepG 2细胞膜上的表皮生长因子受体区。BBR和THP对HepG 2细胞增殖的抑制作用呈浓度依赖性（p < 0.05）。结果表明，所构建的二维HepG 2/CMC/整体柱/飞行时间质谱系统具有较强的识别能力和快速分析能力，可用于复杂样品的药物筛选，也可应用于其他生物色谱模型。(C)2012爱思唯尔有限公司版权所有。
Cell membrane chromatography (CMC) is a biological affinity chromatographic method using specific cell membrane as stationary phase. It has been proved to be a practical tool for investigating binding interactions between drugs and membrane receptors. In this study, a novel comprehensive two-dimensional (2D) chromatography approach was established for screening anti-tumor components from herbal medicines (HMs). HepG2/CMC model was first developed and applied as the first dimensional column. Using an automatic ten-port switching valve equipped with two sample loops, the fractions of the first-dimension were introduced in the second-dimension consists of a monolithic column and a time-of-flight mass spectrometry (TOFMS) with high resolving ability. Based on the stability, selectivity and suitability assays of the HepG2/CMC/monolithic column/TOFMS system, berberine (BBR) and tetrahydropalmatine (THP) from Cortex phellodendri amurensis, oxymatrine and matrine from Radix sophorae flavescentis were screened and identified as potential active components. The competitive displacement assay suggested that the four components could act on epidermal growth factor receptor region on the HepG2 cell membrane in similar manner of gefitinib. Furthermore, their inhibiting effects on cell proliferation in vitro were also confirmed and, BBR and THP showed concentration dependently inhibitory ability on HepG2 cell proliferation (p < 0.05). The result demonstrated that the proposed comprehensive 2D HepG2/CMC/monolithic column/TOFMS system has the advantages of strong recognition and rapid analysis abilities for the total screening procedure, which will be selectable and practical in drug discovery from complex HM samples and can also be applied to other biochromatography models. (C) 2012 Elsevier B.V. All rights reserved.