title: HepG2 cells biospecific extraction and HPLC-ESI-MS analysis for screening potential antiatherosclerotic active components in Bupeuri radix.
abstract: 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.
authors.affiliation: 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.
authors.forename: Shuqiang
authors.initials: S
authors.lastname: Liu
authors.affiliation: School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
authors.forename: Zhibin
authors.initials: Z
authors.lastname: Tan
authors.affiliation: School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
authors.forename: Pingting
authors.initials: P
authors.lastname: Li
authors.affiliation: School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Gao
authors.affiliation: School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
authors.forename: Yuaner
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: School of Chinese Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China. Electronic address: jingelwang@gzucm.edu.cn.
authors.forename: Shuling
authors.initials: S
authors.lastname: Wang
mesh_terms: D050197:Atherosclerosis / Q000188:drug therapy*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000494:pharmacology*; D056945:Hep G2 Cells; D006801:Humans; D008516:Medicine, Chinese Traditional; D009828:Oleanolic Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D018517:Plant Roots / Q000737:chemistry*; D012503:Saponins / Q000737:chemistry / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D004365:Drugs, Chinese Herbal; D012503:Saponins; D009828:Oleanolic Acid; C025759:saikosaponin D
keywords: Antiatherosclerotic active components; Bupeuri radix; Cellular uptake of DiI-HDL; HPLC-ESI-MS; HepG2 cells biospecific extraction