On-line comprehensive two-dimensional HepG2 cell membrane chromatographic analysis system for charactering anti-hepatoma components from rat serum after oral administration of Radix scutellariae: A strategy for rapid screening active compounds in vivo
On-line comprehensive two-dimensional HepG2 cell membrane chromatographic analysis system for charactering anti-hepatoma components from rat serum after oral administration of Radix scutellariae: A strategy for rapid screening active compounds in vivo
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在线综合二维HepG2细胞膜色谱分析系统表征口服黄芩后大鼠血清抗肝癌成分：体内活性化合物快速筛选策略。
DOI：
10.1016/j.jpba.2015.10.013
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发表时间：
2016-01-25
影响因子：
3.4
通讯作者：
Zhu, Zhenyu
中科院分区：
文献类型：
--
作者：
Jia, Dan;Chen, Xiaofei;Zhu, Zhenyu
细胞膜色谱(CMC)是一种表征药物与膜受体相互作用的生物亲和层析技术，已被广泛应用于从中草药等复杂样品中筛选活性成分。然而，由于其相对较高的检测下限(LOD)和基质干扰，它从未在体内应用过。本研究建立了一种新型的在线综合二维HepG2/CMC/富集柱/高效液相色谱/飞行时间质谱仪系统，用于快速筛选大鼠灌胃给药后含药血清中潜在的抗肝癌活性成分。利用MatLab自编程序，建立了一种消除血清中内源性物质干扰的基质干扰分析方法。在HepG2/CMC柱上，黄芩素、汉黄素、白杨素、冬青素A、新黄芩素和白藜芦素被显著保留。首次从含药血清中筛选出汉黄藤素、新黄芩素A和新黄芩素3个潜在活性成分。Kit-8细胞计数实验表明，汉黄藤素、白杨素A和白杨素在12.5~200 mU M浓度范围内对HepG2细胞有较强的抑制作用，且呈剂量依赖关系(p
Cell membrane chromatography (CMC) is a bioaffinity chromatography technique for characterizing interactions between drugs and membrane receptors and has been widely used to screen active components from complex samples such as herbal medicines (HMs). However, it has never been applied in vivo due to its relatively high limit of detection (LOD) and the matrix interferences. In this study, a novel on-line comprehensive two-dimensional HepG2/CMC/enrich columns/high performance liquid chromatography/time-of-flight mass spectrometry system was developed to rapidly screen potential anti-hepatoma components from drug-containing serum of rats after oral administration of Radix scutellariae. A matrix interference deduction method with a home-written program in MATLAB was developed, which could successfully eliminate the interference of endogenous substances in serum. Baicalein, wogonin, chrysin, oroxylin A, neobaicalein and rivularin from Radix scutellariae extraction were significantly retained in the HepG2/CMC column. Three potential active components, wogonin, oroxylin A and neobaicalein were firstly screened from the drug-containing serum as well. The cell counting kit-8 assay demonstrated that wogonin, oroxylin A and chrysin showed high inhibitory activities in a dosedependent manner on HepG2 cells at the concentration of 12.5-200 mu M (p