用高表达EGFR细胞膜色谱-HPLC/MS联用快速筛选独活中抗EGFR活性成分

被引:30
作者
王嗣岑 [1 ,2 ]
孙萌 [1 ,2 ]
张彦民 [1 ,2 ]
张杰 [1 ,2 ]
贺浪冲 [1 ,2 ]
机构
[1] 环境与疾病相关基因教育部重点实验室
[2] 西安交通大学医学院
基金
国家自然科学基金重点项目;
关键词
EGFR/细胞膜色谱二维色谱; 快速筛选; 独活; 抗EGFR活性;
D O I
暂无
中图分类号
R284.1 [化学分析与鉴定];
学科分类号
100804 [中药化学];
摘要
报道了用高表达表皮生长因子受体细胞膜色谱与高效液相色谱/质谱在线联用方法(EGFR/CMC-online-HPLC/MS)快速筛选发现中药独活中的活性成分.实验中,采用高表达EGFR的细胞膜制备色谱固定相,建立EGFR/细胞膜色谱(EGFR/CMC)模型,利用柱切换和固相萃取技术,将EGFR/CMC模型与高效液相色谱/质谱(HPLC/MS)在线联用,构成一种新的可同时"识别-鉴定"目标成分的二维色谱系统,并应用于快速筛选独活中具有抗EGFR活性的目标成分.结果发现独活中的蛇床子素具有与对照药物达沙替尼类似的色谱保留特性,能够作用于EGFR;同时MTT及Elisa分析实验证实蛇床子素对HEK293EGFR细胞增殖及EGFR表达均有抑制作用.本文建立的EGFR/CMC-online-HPLC/MS二维色谱方法,可以选择性地从中药复杂体系中快速"识别-鉴定"目标组分,且筛选结果与特定生物效应显著相关.
引用
收藏
页码:750 / 756
页数:7
相关论文
共 18 条
[1]
细胞培养.[M].司徒镇强;吴军正主编;.世界图书出版公司西安分公司.1996,
[2]
Screening anti-inflammatory components from Chinese traditional medicines using a peritoneal macrophage/cell membrane chromatography-offline-GC/MS method [J].
Wang, Changhe ;
He, Langchong ;
Wang, Nan ;
Liu, Fang .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2009, 877 (27) :3019-3024
[3]
A vascular smooth muscle/cell membrane chromatography–offline-gas chromatography/mass spectrometry method for recognition; separation and identification of active components from traditional Chinese medicines.[J].Xiaofang Hou;Mingzhe Zhou;Qiao Jiang;Sicen Wang;Langchong He.Journal of Chromatography A.2009, 42
[4]
Fluorenone alkaloid from Caulophyllum robustum Maxim. with anti-myocardial ischemia activity [J].
Wang, Sicen ;
Wen, Binyu ;
Wang, Na ;
Liu, Juntian ;
He, Langchong .
ARCHIVES OF PHARMACAL RESEARCH, 2009, 32 (04) :521-526
[5]
Single-molecule Imaging and fluorescence lifetime imaging microscopy show different structures for high-and low-affinity epidermal growth factor receptors in A431 cells [J].
Webb, Stephen E. D. ;
Roberts, Selene K. ;
Needham, Sarah R. ;
Tynan, Christopher J. ;
Rolfe, Daniel J. ;
Winn, Martyn D. ;
Clarke, David T. ;
Barraclough, Roger ;
Martin-Fernandez, Marisa L. .
BIOPHYSICAL JOURNAL, 2008, 94 (03) :803-819
[6]
Screening for the anti-inflammatory activity of fractions and compounds from Atractylodes macrocephala koidz [J].
Li, Cui-Qin ;
He, Lang-Chong ;
Dong, Hai-Yan ;
Jin, Ju-Qing .
JOURNAL OF ETHNOPHARMACOLOGY, 2007, 114 (02) :212-217
[7]
Comparison of alpha1-adrenergic receptor cell-membrane stationary phases prepared from expressed cell line and from rabbit hepatocytes [J].
Wang, Yu ;
Yuan, Bingxiang ;
Deng, Xiuling ;
He, Langchong ;
Wang, Sicen ;
Zhang, Youyi ;
Han, Qide .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2006, 386 (7-8) :2003-2011
[8]
Screening, analysis and in vitro vasodilatation of effective components from Ligusticum Chuanxiong [J].
Liang, MJ ;
He, LC ;
Yang, GD .
LIFE SCIENCES, 2005, 78 (02) :128-133
[9]
The preparation of HEK293 α1A or HEK293 α1B cell membrane stationary phase and the chromatographic affinity study of ligands of α1 adrenoceptor [J].
Wang, Y ;
Yuan, BX ;
Deng, XL ;
He, LC ;
Zhang, YY ;
Han, QD .
ANALYTICAL BIOCHEMISTRY, 2005, 339 (02) :198-205
[10]
Comparison of determination of drug-muscarinic receptor affinity by cell-membrane chromatography and by radioligand-binding assay with the cerebrum membrane of the rat [J].
Yuan, BX ;
Hou, J ;
Yang, GD ;
Zhao, LM ;
He, LC .
CHROMATOGRAPHIA, 2005, 61 (7-8) :381-384