title: Combining human periodontal ligament cell membrane chromatography with online HPLC/MS for screening osteoplastic active compounds from Coptidis Rhizoma.
abstract: We have developed an online analytical method that combines human periodontal ligament cell membrane chromatography (hPDLC/CMC) with high-performance liquid chromatography and mass spectrometry (LC/MS) for recognizing and identifying osteoplastic active components from Coptidis Rhizoma. Retention fractions on hPDLC/CMC were enriched onto an enrichment column and the components were directly analyzed by combining a 10-port column switcher with an LC/MS system for separation and preliminary identification. Using simvastatin (SIM) as a positive control, berberine from Coptidis Rhizoma was identified as the active component which could act on the hPDLC. The MTT colorimetric assay, alkaline phosphatase (ALP) activity, and staining tests revealed that berberine could promote hPDLC growth, increase the secretion of ALP in the culture medium, and enhance the formation of mineralized nodule, thus it is a potential osteoplastic ingredient. This hPDLC/CMC-online-LC/MS method can be applied for screening active components acting on hPDLC from traditional Chinese medicines exemplified by Coptidis Rhizoma and will be of great utility in drug discovery using natural medicinal herbs as a source of leading compounds.
authors.affiliation: Stomatological Hospital, Xi'an Jiaotong University, Xi'an 710 004, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Liu
authors.forename: Jin
authors.initials: J
authors.lastname: Yang
authors.forename: Sicen
authors.initials: S
authors.lastname: Wang
authors.forename: Junyi
authors.initials: J
authors.lastname: Sun
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Shi
authors.forename: Guozhou
authors.initials: G
authors.lastname: Rao
authors.forename: Ang
authors.initials: A
authors.lastname: Li
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Gou
mesh_terms: D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D000704:Analysis of Variance; D001599:Berberine / Q000302:isolation & purification / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D039323:Coptis / Q000737:chemistry; D000089042:Coptis chinensis; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D010513:Periodontal Ligament / Q000737:chemistry / Q000166:cytology*; D019821:Simvastatin / Q000494:pharmacology
chemical_list: D004365:Drugs, Chinese Herbal; D001599:Berberine; D019821:Simvastatin; C107393:Coptidis rhizoma extract; D000469:Alkaline Phosphatase