基于细胞膜色谱法的新疆准噶尔乌头中抗EGFR活性成分筛选  

Screening of Anti-EGFR Components from Aconitum Soongaricum Stapf. in Xinjiang Province Based on Cell Membrane Chromatography

作　　者：木卡代斯.斯依提 林园园 张江[1] 赵翡翠[3] Mukadaisi Siyiti;Lin Yuanyuan;Zhang Jiang;Zhao Feicui(College of Traditional Chinese Medicine, Xinjiang Medical University, Urumqi 830000, China;School of Pharmacy, Xi＇ an Jiaotong University;Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University)

机构地区：[1]新疆医科大学中医学院,乌鲁木齐830000 [2]西安交通大学药学院 [3]新疆医科大学附属中医医院药学部

出　　处：《中国药师》2018年第5期766-771,共6页China Pharmacist

基　　金：国家自然科学基金项目(编号:81460603)

摘　　要：目的:从准噶尔乌头中筛选抗EGFR作用的活性成分,并考察筛选成分对HEK293细胞和EGFR/HEK293高表达细胞株的抑制活性。方法:用高表达表皮生长因子受体细胞膜色谱(EGFR/CMC)通过十通切换阀与高效液相色谱(HPLC)构成二维在线联用色谱系统,将CMC上保留的组分进行接液,用离线MS来鉴定准噶尔乌头中抗EGFR活性成分;利用分子模拟对接实验分析活性成分与受体的相互作用方式;体外用MTT实验证实活性组分对HEK293细胞和EGFR/HEK293高表达细胞株的抑制作用。结果:乌头碱、12-表-欧乌碱、准噶尔乌头碱为作用于EGFR的有效成分,并具有与对照药物吉非替尼类似的色谱保留特性;分子模拟对接实验表明这些成分与EGFR的相互作用方式与阳性药吉非替尼相似;MTT实验证实在浓度为0.4~50μmol·L^(-1)范围内乌头碱,12-表-欧乌碱,准噶尔乌头碱对HEK293细胞增殖与EGFR高表达的体外抑制作用具有剂量依赖性。结论:筛选的结果与生物效应相一致;在线EGFR/CMC-HPLC-MS二维色谱系统在中药复杂体系中快速筛选发现活性物和药物先导物方面具有良好的应用前景,同时充分展现了细胞膜色谱法操作简便,快速,高效灵敏的优势。Objective： To screen anti-EGFR components from Aconitum soongaricum Stapf. and investigate their inhibitory effect on HEK293 ceils and EGFR/HEK293 high expression cell lines. Methods： A two-dimensional online system （EGFR/CMC-HPLC-MS） was built to screen anti-EGFR active compounds from Aconitum soongaricum Stapf. ; molecular docking assay was performed to determine the binding affinity of the components to EGFR ; MTT was used to confirm the inhibition effect of the screened compounds on HEK293 cells and EGFR/HEK293 high expression cell lines. Results： The screening results showed that aconitine, 12-epi-napellin and songorine were the active components acting on EGFR like gefitinib; molecular docking assay showed the targeting components acting on EGFR in the similar manner with gefitinib used as a positive control drug; MTT assay confirmed the screened components had inhibitory effects on HEK293 cells and EGFR/HEK293 high expression cell lines in a dose-dependent manner within the dose range of 0.4-50.0 μmol · L-1. Conclusion： The screening results are apparently consistent with the pharmacological effect. The online EGFR/CMC-HPLC-MS method has a good application prospect in the rapid screening of the active compounds in the complex system of traditional Chinese medicines, and fully demonstrates such advantages of cell membrane chromatography as simple and rapid with high efficiency.

关 键 词：细胞膜色谱 二维色谱 新疆准噶尔乌头 抗EGFR活性 

分 类 号：R284[医药卫生—中药学]