基于HUVEC细胞膜色谱的莱菔子中抗血管性疾病活性成分筛选及验证  

Screening and validation of anti-vascular disease active components from Semen raphani based on HUVEC cell membrane chromatography

作　　者：李华妮 刘长河[1,2] 菅单单 陈胜虎 葛文静 张雪侠 LI Hua-ni;LIU Chang-he;JIAN Dan-dan;CHEN Sheng-hu;GE Wen-jing;ZHANG Xue-xia(Henan Integrative Medicine Hospital/Henan Provincial Academy of Traditional Chinese Medicine and Pharmacy,Zhengzhou 450004,China;Henan Engineering Research Center of Traditional Chinese Medicine Preparation,Zhengzhou 450004,China;Pharmaceutical College of Henan University,Kaifeng 475004,China)

机构地区：[1]河南省中西医结合医院河南省中医药研究院,郑州450004 [2]河南省中药制剂工程技术研究中心,郑州450004 [3]河南大学药学院,开封475004

出　　处：《药物分析杂志》2024年第6期929-937,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：河南省自然科学基金(232300421318);河南省中医药科学研究专项课题(20-21ZY2262);河南省中医药科学研究专项课题(2022ZY1148);河南省基本科研业务费(2304574)。

摘　　要：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HUVEC,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。Objective:To establish a method for determination of potential anti-vascular disease active components 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 reference 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 protein levels decreased and Bax protein levels increased,with statistical significance(P<0.05).Conclusion:The method 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.

关 键 词：莱菔子 细胞膜色谱 芥子酸 人脐静脉内皮细胞 

分 类 号：R917[医药卫生—药物分析学]