Liu Shuqiang, Tan Zhibin, Li Pingting, Gao Xiaoling, Zeng Yuaner, Wang Shuling
School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China; International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
J Pharm Biomed Anal. 2016 Mar 20;121:56-62. doi: 10.1016/j.jpba.2016.01.010. Epub 2016 Jan 8.
HepG2 cells biospecific extraction method and high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) analysis was proposed for screening of potential antiatherosclerotic active components in Bupeuri radix, a well-known Traditional Chinese Medicine (TCM). The hypothesis suggested that when cells are incubated together with the extracts of TCM, the potential bioactive components in the TCM should selectively combine with the receptor or channel of HepG2 cells, then the eluate which contained biospecific component binding to HepG2 cells was identified using HPLC-ESI-MS analysis. The potential bioactive components of Bupeuri radix were investigated using the proposed approach. Five compounds in the saikosaponins of Bupeuri radix were detected as these components selectively combined with HepG2 cells, among these compounds, two potentially bioactive compounds namely saikosaponin b1 and saikosaponin b2 (SSb2) were identified by comparing with the chromatography of the standard sample and analysis of the structural clearance characterization of MS. Then SSb2 was used to assess the uptake of DiI-high density lipoprotein (HDL) in HepG2 cells for antiatherosclerotic activity. The results have showed that SSb2, with indicated concentrations (5, 15, 25, and 40 μM) could remarkably uptake dioctadecylindocarbocyanine labeled- (DiI) -HDL in HepG2 cells (Vs control group, *P<0.01). In conclusion, the application of HepG2 biospecific extraction coupled with HPLC-ESI-MS analysis is a rapid, convenient, and reliable method for screening potential bioactive components in TCM and SSb2 may be a valuable novel drug agent for the treatment of atherosclerosis.
提出了一种利用人肝癌细胞系HepG2细胞生物特异性提取方法和高效液相色谱-电喷雾电离-质谱联用（HPLC-ESI-MS）分析法，筛选著名中药柴胡中潜在抗动脉粥样硬化活性成分的研究。该假说认为，当细胞与中药提取物共同孵育时，中药中的潜在生物活性成分应能选择性地与HepG2细胞的受体或通道结合，然后利用HPLC-ESI-MS分析法鉴定含有与HepG2细胞结合的生物特异性成分的洗脱液。采用该方法对柴胡的潜在生物活性成分进行了研究。检测到柴胡皂苷中的5种化合物能选择性地与HepG2细胞结合，通过与标准样品色谱图对比及质谱结构解析，鉴定出两种潜在生物活性化合物，即柴胡皂苷b1和柴胡皂苷b2（SSb2）。随后，利用SSb2评估其对HepG2细胞摄取DiI标记的高密度脂蛋白（HDL）的影响，以研究其抗动脉粥样硬化活性。结果表明，不同浓度（5、15、25和40 μM）的SSb2均可显著促进HepG2细胞摄取二辛基碳菁染料标记的HDL（与对照组相比，*P<0.01）。综上所述，HepG2细胞生物特异性提取结合HPLC-ESI-MS分析方法是一种快速、便捷、可靠的筛选中药潜在生物活性成分的方法，SSb2可能是一种有价值的治疗动脉粥样硬化的新型药物。
