title: A novel semisynthetic molecule icaritin stimulates osteogenic differentiation and inhibits adipogenesis of mesenchymal stem cells.
abstract: BACKGROUND
  We previously reported that the constitutional flavonoid glycosides derived from herb Epimedium (EF, composed of seven flavonoid compounds with common nuclear stem) exerted beneficial effects on the bone, including promoting bone formation and inhibiting bone marrow fat deposition. Recent in vivo study showed that Icaritin was a common metabolite of these constitutional flavonoid glycosides, indicating that Icaritin is a bioactive compound. The present study was designed to investigate whether Icaritin could promote osteogenic differentiation and suppress adipogenic differentiation of marrow mesenchymal stem cells (MSCs).
  
  
  METHODS
  Primary MSCs were harvested from adult mice and exposed to Icaritin to evaluate whether it could promote osteogenesis and suppress adipogenesis using the following assays: determination of alkaline phosphatase (ALP) activity and mineralization; mRNA expression of osteogenic differentiation marker Runx2; osteocalcin and bone sialoprotein (BSP) by RT-PCR; quantification of adipocyte-like cells by Oil Red O staining assay and mRNA expression for adipogenic differentiation markers peroxisome proliferator-activated receptor gamma (PPARγ); adipocyte fatty acid binding protein (aP2) and lipoprotein lipase (LPL) by RT-PCR. For the underlying mechanism, glycogen synthase kinase-3beta (GSK3β) and β-catenin were also explored by western blotting.
  
  
  RESULTS
  Icaritin promoted osteogenic differentiation and maturation of MSCs as indicated by increased mRNA expression for Runx2, osteocalcin and BSP, and enhanced ALP activity and mineralization; Icaritin inhibited adipogenic differentiation, as indicated by decreased mRNA expression for PPARγ, LPL, aP2, and suppressed formation of adipocyte-like cells; Icaritin inactivated GSK3β and suppressed PPARγ expression when promoting osteogenesis and suppressing adipogenesis of MSCs.
  
  
  CONCLUSIONS
  This was the first study demonstrating that the novel semisynthetic molecule Icaritin could stimulate osteogenic differentiation and inhibit adipogenesis of MSCs, which was associated with the suppression of GSK3β and PPARγ.
authors: Department of Endocrinology and Metabolism, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China. shenghui@tongji.edu.cn
authors: Hui
authors: H
authors: Sheng
authors: Xue-fei
authors: XF
authors: Rui
authors: Chun-Jun
authors: CJ
authors: Sheng
authors: Wen-Jun
authors: WJ
authors: Li
authors: Xiao-Yun
authors: XY
authors: Cheng
authors: Navina Priya
authors: NP
authors: Jhummon
authors: Yong-Chun
authors: YC
authors: Yu
authors: Shen
authors: S
authors: Qu
authors: Ge
authors: G
authors: Zhang
authors: Ling
authors: L
authors: Qin
mesh_terms: D050156:Adipogenesis / Q000187:drug effects; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D005419:Flavonoids / Q000008:administration & dosage* / Q000138:chemical synthesis; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D010012:Osteogenesis / Q000187:drug effects*; D047495:PPAR gamma / Q000378:metabolism
chemical_list: D005419:Flavonoids; D047495:PPAR gamma; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D038362:Glycogen Synthase Kinase 3; C499403:icaritin
keywords: Adipogenesis; GSK3β; Icaritin; Mesenchymal stem cells; Osteogenesis; Osteoporosis.
references: Tissue Eng. 2006 Jul;12(7):1753-61
references: 16889506
references: PLoS One. 2007 Sep 05;2(9):e837
references: 17786208
references: Bone. 2004 Mar;34(3):393-401
references: 15003787
references: Radiology. 2006 Dec;241(3):831-8
references: 17053202
references: Bone. 2006 Jun;38(6):818-25
references: 16413840
references: Clin Orthop Relat Res. 2005 Sep;438:260-5
references: 16131900
references: J Orthop Surg Res. 2007 Oct 15;2:15
references: 17937789
references: Blood Cells Mol Dis. 2001 May-Jun;27(3):677-90
references: 11482883
references: Diabetes. 2000 Jun;49(6):904-11
references: 10866041
references: Curr Opin Pharmacol. 2004 Jun;4(3):290-4
references: 15140422
references: J Clin Invest. 2008 Feb;118(2):491-504
references: 18219387
references: Gene. 2005 Mar 28;348:157-65
references: 15777692
references: Mol Cell. 1999 Oct;4(4):611-7
references: 10549292
references: J Bone Miner Res. 2002 Mar;17(3):394-405
references: 11878304
references: J Cell Biochem. 2008 Mar 1;103(4):1054-65
references: 17973271
references: Arthritis Rheum. 2009 Oct;60(10):2966-77
references: 19790047
references: Curr Osteoporos Rep. 2007 Jun;5(2):73-80
references: 17521509
references: Life Sci. 2007 Aug 16;81(10):832-40
references: 17764702
references: PLoS One. 2009 Jul 29;4(7):e6419
references: 19641620
references: Bone. 2007 Mar;40(3):685-92
references: 17188589
references: J Endocrinol. 1999 Apr;161(1):89-97
references: 10194532
references: J Cell Biochem. 2006 May 15;98(2):251-66
references: 16479589
references: Stem Cells. 2007 Feb;25(2):364-70
references: 17068184
references: J Bone Miner Res. 2007 Jul;22(7):1072-9
references: 17419678
references: Exp Cell Res. 2004 May 1;295(2):395-406
references: 15093739
references: J Cell Biochem. 2007 Feb 1;100(2):464-73
references: 16927271
references: Cell. 1997 May 30;89(5):755-64
references: 9182763
references: J Reprod Dev. 2007 Jun;53(3):563-72
references: 17272924