目的:构建人脐静脉细胞膜色谱(HUVEC/CMC)模型,并将其应用于莱菔子中抗血管性疾病活性成分的快速筛选.方法:采用HUVEC/CMC模型二维在线联用HPLC-ESI IT TOF MS系统对莱菔子活性成分进行分离、筛选和鉴定,并将筛选的活性成分进一步作用于氧化低密度脂蛋白(ox-LDL)诱导的HU-VEC,验证其保护作用.结果:利用所建立的方法从莱菔子中共筛选出2个保留组分,通过与对照品比对,鉴定出1个成分为芥子酸.与模型组比较,芥子酸低、中、高预处理组细胞存活率显著增加,细胞中ICAM-1和VCAM-1的含量下降,并呈剂量依赖性;Bcl-2蛋白表达水平下降、Bax蛋白表达升高,具有统计学意义(P＜0.05).结论:构建的HUVEC/CMC在线联用HPLC-ESI IT TOF MS系统可用于快速筛选复杂中药体系中活性成分,为细胞膜色谱的应用和莱菔子的开发提供了参考.
Screening and validation of anti-vascular disease active components from Semen raphani based on HUVEC cell membrane chromatography
Objective:To establish a method for determination of potential anti-vascular disease active compo-nents of Semen raphani based on cell membrane chromatographic(CMC)model of human umbilical vein cell(HUVEC).Methods:The HUVEC cell membrane chromatography coupled with HPLC-ESI IT TOF MS system were used to screen the active components of Semen raphani.The selected active components were further applied to ox-LDL induced HUVEC to verify their protective effects.Results:Two retained components were selected from Semen raphani by this method.One component was identified as erucinic acid by comparing with the refer-ence material.Compared with the model group,the cell survival rates of the erucinic acid pretreatment groups increased significantly.The amount of ICAM-1 and VCAM-1 decreased in a dose-dependent manner.Bcl-2 pro-tein levels decreased and Bax protein levels increased,with statistical significance(P＜0.05).Conclusion:The meth-od can rapidly obtain active ingredients from complex traditional Chinese medicines.It provides a reference for the application of cell membrane chromatography and the development of Semen raphani.
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