title: Coordinate regulatory osteogenesis effects of icariin, timosaponin B II and ferulic acid from traditional Chinese medicine formulas on UMR-106 osteoblastic cells and osteoblasts in neonatal rat calvaria cultures.
abstract: BACKGROUND
  Icariin (I), ferulic acid (F) and timosaponin B II (T) derived respectively from the leaf of Epimedium brevicornu Maxim (EBM, Berberidaceae), rhizome of Anemarrhena asphodeloides Bunge (AAB, Liliaceae) and root of Angelica sinensis (Oliv.) Diels (ASD, Umbelliferae) are included in several traditional Chinese medicine (TCM) formulas for the treatment of osteoporosis. In addition, the medicinal materials and chemical constituents in many traditional Chinese formulas have been shown to have potential synergistic, additive and antagonistic effects.
  
  
  OBJECTIVE
  To explore the action mechanism and interactions between I, T and F as bone anabolic ingredients on osteoblasts, and fully understand their action mechanism and rationality of the formula design.
  
  
  METHODS
  An osteoporotic model was established in bilaterally ovariectomized mice. Bone mineral density (BMD), bone mineral content (BMC) and serum biochemical parameters including alkaline phosphatase (ALP), tartrate resistant acid phosphatase (TRAP), osteoprotegerin (OPG) and deoxypyridinoline cross-links (DPD) were measured to evaluate the effects of I, T or F alone and their combinations on osteoporotic mice. UMR-106 osteoblastic cells and primary osteoblasts in neonatal rat calvarias were used to evaluate the osteogenesis effect. The immunohistochemical method and Western-blot analysis were used to detect the expression of critical proteins in the process of proliferation and differentiation of osteoblasts.
  
  
  RESULTS
  IFT combinations enhanced the therapeutic effect without increasing the adverse effects on osteoporotic mice, synergistically increased the osteoblast proliferation, ALP activity and mineralized nodule formation, and promoted the expression of bone matrix by regulating BMP and Wnt/β-catenin signaling pathways in osteoblasts.
  
  
  CONCLUSIONS
  IFT combinations reinforced the therapeutic effect on osteoporosis by modulating multi-signaling pathways and action targets.
authors.affiliation: School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China; XiaMen ZhongKe New Biotechnology Co., Ltd., Xiamen 361000, China. Electronic address: Lm0836@163.com.
authors.forename: Mei
authors.initials: M
authors.lastname: Li
authors.affiliation: School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China. Electronic address: daniell_znd@163.com.
authors.forename: Nai-Dan
authors.initials: ND
authors.lastname: Zhang
authors.affiliation: Department of Pharmaceuticals, PLA 102 Hospital, Changzhou, 213003 China. Electronic address: wangsbox@sina.com.
authors.forename: Yin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China. Electronic address: than927@163.com.
authors.forename: Ting
authors.initials: T
authors.lastname: Han
authors.affiliation: School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China. Electronic address: msjyp@163.com.
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Jiang
authors.affiliation: School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, UK. Electronic address: K.Rahman@ljmu.ac.uk.
authors.forename: Khalid
authors.initials: K
authors.lastname: Rahman
authors.affiliation: School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China. Electronic address: qinluping@126.com.
authors.forename: Lu-Ping
authors.initials: LP
authors.lastname: Qin
authors.affiliation: School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China. Electronic address: zqy1965@163.com.
authors.forename: Hai-Liang
authors.initials: HL
authors.lastname: Xin
authors.affiliation: School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China. Electronic address: hailiangxin@163.com.
authors.forename: Qiao-Yan
authors.initials: QY
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003373:Coumaric Acids / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D005260:Female; D005419:Flavonoids / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D005786:Gene Expression Regulation; D051379:Mice; D015394:Molecular Structure; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects*; D010024:Osteoporosis / Q000188:drug therapy; D051381:Rats; D012503:Saponins / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D013256:Steroids / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D003373:Coumaric Acids; D004365:Drugs, Chinese Herbal; D005419:Flavonoids; D012503:Saponins; D013256:Steroids; C523027:timosaponin B-II; C004999:ferulic acid; D002118:Calcium; C056599:icariin
keywords: BMP signaling pathway; Ferulic acid; Ferulic acid (Pubchem CID: 445858); Iacriin (Pubchem CID: 5318997); Icariin; Osteogenic differentiation; Osteoporosis; Timosaponin B II; Timosaponin B II (Pubchem CID: 44575945); Traditional Chinese medicine formula; Wnt/β-Catenin signaling pathway