HepG2 cells biospecific extraction and HPLC-ESI-MS analysis for screening potential antiatherosclerotic active components in Bupeuri radix

• In this study, the potential antiatherosclerotic active components in the Radix Bupeuri extract were studied using HepG2Cells biospecific extraction and high performance liquid chromatography-electron spray ionization-mass spectrometry (HPLC-ESI-MS).
• Five compounds in the saikosaponins of Radix Bupeuri (SSRB) were detected as the components selectively combined with HepG2 cells, among these compounds, two potential bioactive compounds, saikosaponin b1 (SSb1) and saikosaponin b2 (SSb2) were identified by comparing with the chromatography of the standard sample and analysis of the structure clearage characterization of mass spectrometry.
• To test whether SSb2 enhanced the selective uptake of lipids from HDL by fluorescence-labeled DiI-HDL uptake after 4 h incubation with HepG2 cells was measured in the presence of indicated concentration of SSb2 or vehicle (0.1% DMSO).
• In conclusion, the application of HepG2 biospecific extraction coupled with HPLC-ESI-MS analysis is a rapid, convenient and reliable method for screening potent active components in TCM and SSb2 may be of value as a novel drug agent for the atherosclerosis.

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.

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