title: Inhibitory effect of Ecliptae herba extract and its component wedelolactone on pre-osteoclastic proliferation and differentiation.
abstract: BACKGROUND
  Ecliptae herba, also known as "Mo-Han-Lian", has long been used in China to nourish Kidney and thereafter strengthen bones. Accumulating evidence indicates that extracts of Ecliptae herba have antiosteoporotic effect. However, the effective compounds and cellular mode of action are still unclear. To investigate the effect of ethyl acetate extract of Ecliptae herba (EAE) and its component wedelolactone on proliferation and differentiation of preosteoclastic RAW264.7 cells as well as proliferation of bone marrow stromal cells (BMSC).
  
  
  METHODS
  RAW264.7 and BMSC were examined for proliferation by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method. Tartrate-resistant acid phosphatase (TRAP) activity of RAW264.7 was measured by using p-nitrophenyl sodium phosphate (pNPP) assay after the cells were treated with 30ng/ml receptor activator for nuclear factor-κ B ligand (RANKL) plus various concentrations of EAE, wedelolactone or alendronate. The formation of multinucleated TRAP-positive RAW264.7 cells was observed by using a TRAP-staining kit.
  
  
  RESULTS
  Treatment of RAW264.7 cells with EAE at high doses (20µg/ml and 40µg/ml) or wedelolactone at 10µg/ml resulted in a decrease in proliferation of RAW264.7 cells. Low doses of EAE (5, 10µg/ml) and wedelolactone (2.5µg/ml) inhibited RANKL-induced TRAP activity by 20.3%, 37.9%, and 48.3%. The inhibitory effect of wedelolactone is more potent than that of alendronate, an anti-resorptive drug. Morphological changes revealed that 5µg/ml EAE and 2.5µg/ml wedelolactone reduced the number of multinucleated osteoclast-like cells. At the high doses, EAE (20µg/ml) and wedelolactone (10µg/ml) inhibited the growth of BMSC.
  
  
  CONCLUSIONS
  EAE and its component wedelolactone inhibited osteoclast RAW264.7 proliferation and differentiation at the low doses, but at the high doses, showed cytotoxic effect on BMSC. These results indicated that EAE and wedelolatone might be potential alternative therapy for osteoporosis.
authors.affiliation: College (Institute) of Integrative Medicine, Dalian Medical University, Dalian 116044, China; Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Yan-Qiu
authors.initials: YQ
authors.lastname: Liu
authors.affiliation: The Second Affiliated Hospital, Dalian Medical University, Dalian 116023, Liaoning Province, China; College (Institute) of Integrative Medicine, Dalian Medical University, Dalian 116044, China.
authors.forename: Li-Bin
authors.initials: LB
authors.lastname: Zhan
authors.affiliation: Beijing University of Chinese Medicine, Beijing 100029, China.
authors.forename: Tiegang
authors.initials: T
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Meng-Chun
authors.initials: MC
authors.lastname: Cheng
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. Electronic address: hbxiao@dicp.ac.cn.
authors.forename: Hong-Bin
authors.initials: HB
authors.lastname: Xiao
mesh_terms: D019386:Alendronate / Q000494:pharmacology; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D003374:Coumarins / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D036601:Eclipta / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology
chemical_list: D003374:Coumarins; D010936:Plant Extracts; C051122:wedelolactone; D019386:Alendronate
keywords: Acetonitrile (PubChem CID: 6342); Alendronate (PubChem CID: 2088); DMSO (PubChem CID: 679); Differentiation; Ecliptae herba; Formic acid (PubChem CID: 284); Naphthol AS-MX phosphate (PubChem CID: 74136); Osteoclasts; Penicillin (PubChem CID: 5904); Streptomycin (PubChem CID: 19649); Wedelolactone; Wedelolactone (PubChem CID: 5281813); l-glutamine (PubChem CID: 5961); p-nitrophenyl sodium phosphate (PubChem CID: 378)