title: Dioscin promotes osteoblastic proliferation and differentiation via Lrp5 and ER pathway in mouse and human osteoblast-like cell lines.
abstract: BACKGROUND
  Dioscin, a typical steroid saponin, is isolated from Dioscorea nipponica Makino and Dioscorea zingiberensis Wright. It has estrogenic activity and many studies have also reported that dioscorea plants have an effect in preventing and treating osteoporosis. However, the molecular mechanisms underlying their effect on osteoporosis treatment are poorly understood. Therefore, the present study aims to investigate the mechanism (s) by which dioscin promotes osteoblastic proliferation and differentiation in mouse pre-osteoblast like MC3T3-E1 cells and human osteoblast-like MG-63 cells.
  
  
  RESULTS
  We found that dioscin (0.25 μg/ml, 0.5 μg/ml, and 1.0 μg/ml) promoted MC3T3-E1 cells and MG-63 cells proliferation and differentiation dose dependently. Western blot analysis results showed that estrogen receptor α (ER-α), estrogen receptor β (ER-β), β-catenin and Bcl-2 protein expression increased after MC3T3-E1 cells were treated with dioscin. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that dioscin could increase the ratio of osteoprotegerin (OPG)/receptor activator of NF-κB ligand (RANKL) and up-regulate the level of Lrp5 and β-catenin. And by RNA interference analysis, we proved that the effect of dioscin increasing the ratio of OPG/RANKL was dependent on Lrp5 pathway. In addition, we also found that these effects of dioscin were abolished by ICI 182, 780 (100 nM), an antagonist of ER, indicating that an ER signaling pathway was also involved. We also found that dioscin (0.25 μg/ml, 0.5 μg/ml, and 1.0 μg/ml) induced MG-63 cells proliferation and differentiation in a dose-dependent manner. Western blot analysis results indicated that ER-α, ER-β and β-catenin protein expression increased after MG-63 cells were treated with dioscin.
  
  
  CONCLUSIONS
  The current study is the first to reveal that dioscin can promote osteoblasts proliferation and differentiation via Lrp5 and ER pathway.
authors.forename: Chunfang
authors.initials: C
authors.lastname: Zhang
authors.forename: Jinyong
authors.initials: J
authors.lastname: Peng
authors.forename: Shan
authors.initials: S
authors.lastname: Wu
authors.forename: Yue
authors.initials: Y
authors.lastname: Jin
authors.forename: Fan
authors.initials: F
authors.lastname: Xia
authors.forename: Changyuan
authors.initials: C
authors.lastname: Wang
authors.forename: Kexin
authors.initials: K
authors.lastname: Liu
authors.affiliation: Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, Dalian, China. sunhuijun@dlmedu.edu.cn.
authors.forename: Huijun
authors.initials: H
authors.lastname: Sun
authors.forename: Mozhen
authors.initials: M
authors.lastname: Liu
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D004144:Diosgenin / Q000008:administration & dosage / Q000031:analogs & derivatives*; D047628:Estrogen Receptor alpha / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D060492:Low Density Lipoprotein Receptor-Related Protein-5 / Q000096:biosynthesis* / Q000378:metabolism; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects; D010024:Osteoporosis / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D053246:Receptor Activator of Nuclear Factor-kappa B / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects
chemical_list: C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D060492:Low Density Lipoprotein Receptor-Related Protein-5; C555483:Lrp5 protein, mouse; D053246:Receptor Activator of Nuclear Factor-kappa B; C019357:dioscin; D004144:Diosgenin
references.citation: BMC Med Genet. 2012 Apr 10;13:26
references.pmid: 22487062
references.citation: Cell. 1997 Apr 18;89(2):309-19
references.pmid: 9108485
references.citation: Cell Physiol Biochem. 2002;12(5-6):359-64
references.pmid: 12438772
references.citation: Biol Pharm Bull. 2002 Feb;25(2):193-6
references.pmid: 11853164
references.citation: Cancer. 2000 Aug 15;89(4):817-25
references.pmid: 10951345
references.citation: Eur J Pharmacol. 2012 Feb 15;676(1-3):26-33
references.pmid: 22192930
references.citation: Fitoterapia. 2012 Dec;83(8):1443-50
references.pmid: 22951288
references.citation: Horm Metab Res. 2005 Oct;37(10):589-92
references.pmid: 16278780
references.citation: J Bone Miner Metab. 2011 Jul;29(4):396-403
references.pmid: 21063740
references.citation: J Cell Biochem. 2006 Aug 1;98(5):1123-9
references.pmid: 16479591
references.citation: Biosci Rep. 2006 Feb;26(1):39-43
references.pmid: 16779666
references.citation: J Endocrinol. 2008 May;197(2):R1-6
references.pmid: 18434348
references.citation: Br J Pharmacol. 2010 Feb;159(4):939-49
references.pmid: 20128811
references.citation: PLoS One. 2012;7(9):e43215
references.pmid: 22984413
references.citation: J Bone Miner Metab. 2013 Jan;31(1):96-101
references.pmid: 22948905
references.citation: Nat Med. 2011 Jun;17(6):684-91
references.pmid: 21602802
references.citation: Int Immunopharmacol. 2011 Jun;11(6):712-8
references.pmid: 21296696
references.citation: Mol Cell Endocrinol. 2012 Feb 26;349(2):180-8
references.pmid: 22040599
references.citation: Dev Cell. 2005 May;8(5):739-50
references.pmid: 15866164
references.citation: Biochem Biophys Res Commun. 2014 Jan 10;443(2):658-65
references.pmid: 24333429
references.citation: Bone. 2012 Jun;50(6):1223-33
references.pmid: 22484100
references.citation: Bone. 2000 May;26(5):423-7
references.pmid: 10773580
references.citation: Evid Based Complement Alternat Med. 2012;2012:938574
references.pmid: 22973408
references.citation: Biol Trace Elem Res. 2011 Dec;144(1-3):1437-48
references.pmid: 21739163
references.citation: Dev Cell. 2002 Apr;2(4):389-406
references.pmid: 11970890
references.citation: Steroids. 2013 Jul;78(7):717-25
references.pmid: 23583603
references.citation: J Surg Res. 2012 May 15;174(2):278-83
references.pmid: 21324392
references.citation: J Cell Biochem. 2009 Nov 1;108(4):896-905
references.pmid: 19795382
references.citation: Exp Cell Res. 2009 Oct 15;315(17):2953-62
references.pmid: 19665018
references.citation: J Cell Biol. 2002 Apr 15;157(2):303-14
references.pmid: 11956231
references.citation: J Bone Miner Metab. 2008;26(3):213-7
references.pmid: 18470660
references.citation: Acta Pharmacol Sin. 2012 Oct;33(10):1277-84
references.pmid: 22902986
references.citation: Environ Toxicol Pharmacol. 2012 Sep;34(2):127-135
references.pmid: 22516057
references.citation: Arch Med Res. 2012 May;43(4):274-82
references.pmid: 22704852
references.citation: J Biol Chem. 2006 Oct 20;281(42):31720-8
references.pmid: 16908522
references.citation: Int J Mol Sci. 2012;13(1):56-70
references.pmid: 22312238
references.citation: Biomark Cancer. 2013 Jul 04;5:13-29
references.pmid: 24179395
references.citation: Phytomedicine. 2012 Apr 15;19(6):551-61
references.pmid: 22397994
references.citation: Fitoterapia. 2010 Dec;81(8):1147-56
references.pmid: 20659537
references.citation: Gene. 2004 Oct 27;341:19-39
references.pmid: 15474285
references.citation: J Biol Chem. 2012 Oct 19;287(43):36168-78
references.pmid: 22936801
references.citation: J Neurosci. 2011 Jun 29;31(26):9620-9
references.pmid: 21715627