title: Puerarin promotes osteogenesis and inhibits adipogenesis in vitro.
abstract: BACKGROUND
  Puerarin (daidzein 8-C-glucoside) has potential on preventing osteoporosis. This study aims to investigate the effects of puerarin on osteogenesis and adipogenesis in vitro.
  
  
  METHODS
  CCK-8 assay, alkaline phosphatase (ALP) activity and Alizarin Red S were used to measure the effects of puerarin on proliferation, osteoblastic differentiation, and mineralization in osteoblast-like MC3T3-E1 cells. The effects of puerarin on adipogenesis were measured by Oil Red O staining and intracellular triglyceride level in preadipocyte 3T3-L1 cells. The mRNA and protein levels of osteogenesis- and adiopogenesis-related factors were detected by qRT-PCR and western blot, respectively. Further, the secreted osteocalcin levels and nuclear translocation of β-catenin were detected by ELISA and immunofluorescence assay, respectively.
  
  
  RESULTS
  As to osteogenesis, puerarin could stimulate proliferation (1 μM, P = 0.012; 10 μM, P = 0.015; 20 μM, P = 0.050), ALP activity (20 μM, P = 0.008) and calcium nodule formation (20 μM, P = 0.011) in a dose-dependent manner. Puerarin (20 μM) promoted osteocalcin secretion (P = 0.004) and the protein expression of both osteopontin (P = 0.001) and osteoprotegerin (P = 0.003). As to adipogenesis, puerarin suppressed adipocytes formation and intracellular triglyceride level (P = 0.001). In addition, puerarin (20 μM) decreased the mRNA and protein levels of CCAAT/enhancer binding protein α (P = 0.001, P = 0.002), proliferator-activated receptor γ (P = 0.005, P = 0.003), and adipocyte lipid-binding protein 4 (P = 0.001, P = 0.001). Moreover, phosphorylation of AKT1-Ser437 (10 μM, P = 0.003; 20 μM, P = 0.007) and GSK-Ser9 (10 μM, P = 0.005; 20 μM, P = 0.003), and the nuclear translocation of β-catenin (10 μM, P = 0.006; 10 μM, P = 0.002) were increased in 3T3-L1 cells treated by puerarin.
  
  
  CONCLUSIONS
  Puerarin promoted osteogenesis and inhibited adipogenesis in vivo, and Akt/GSK-3β/β-catenin signaling pathway was involved in the suppression of adipogenesis.
authors: Translational Medicine R&D Center, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China. xl.wang@siat.ac.cn.
authors: Nan
authors: N
authors: Wang
authors: Xinluan
authors: X
authors: Wang
authors: Wenxiang
authors: W
authors: Cheng
authors: Huijuan
authors: H
authors: Cao
authors: Peng
authors: P
authors: Zhang
authors: Ling
authors: L
authors: Qin
references: J Clin Invest. 1992 Jan;89(1):74-8
references: 1729283
references: J Ethnopharmacol. 2011 Dec 8;138(3):737-40
references: 22027447
references: Annu Rev Cell Dev Biol. 1998;14:59-88
references: 9891778
references: Osteoarthritis Cartilage. 2007 Aug;15(8):894-9
references: 17379543
references: Biochem Biophys Res Commun. 2013 Apr 26;434(1):117-23
references: 23537654
references: Proc Natl Acad Sci U S A. 2005 Mar 1;102(9):3324-9
references: 15728361
references: J Biol Chem. 2012 Oct 19;287(43):36168-78
references: 22936801
references: J Ethnopharmacol. 2005 Jan 4;96(1-2):133-8
references: 15588661
references: Cytokine Growth Factor Rev. 1999 Sep-Dec;10(3-4):255-65
references: 10647780
references: Endocrinology. 1996 Aug;137(8):3590-3
references: 8754791
references: Cell. 1997 May 30;89(5):765-71
references: 9182764
references: Bone. 2000 Aug;27(2):177-84
references: 10913909
references: Clin Chem. 1993 Apr;39(4):648-52
references: 8472360
references: J Biol Chem. 2005 Sep 30;280(39):33132-40
references: 16043491
references: Science. 2000 Aug 11;289(5481):950-3
references: 10937998
references: J Nutr. 2000 Jul;130(7):1675-81
references: 10867035
references: Crit Rev Biochem Mol Biol. 2005 Jul-Aug;40(4):229-42
references: 16126487
references: Planta Med. 1998 Oct;64(7):620-7
references: 17253303
references: Endocr Rev. 1994 Jun;15(3):275-300
references: 8076582
references: Proc Nutr Soc. 2003 Nov;62(4):877-87
references: 15018488
references: Toxicol Appl Pharmacol. 2011 Nov 15;257(1):48-58
references: 21884717
references: Science. 1997 Apr 4;276(5309):71-4
references: 9082988
references: Acta Pharmacol Sin. 2005 Sep;26(9):1081-6
references: 16115375
references: Int J Biochem Cell Biol. 2010 Nov;42(11):1802-7
references: 20691279
references: Proc Natl Acad Sci U S A. 1990 Mar;87(6):2172-6
references: 2315310
references: Life Sci. 2006 Jun 20;79(4):324-30
references: 16472823
references: Nat Rev Endocrinol. 2010 Nov;6(11):629-36
references: 20820194
references: J Bone Miner Res. 2004 May;19(5):853-61
references: 15068509
references: J Bone Miner Res. 2000 Nov;15(11):2084-94
references: 11092391
references: J Nutr. 2000 Dec;130(12):3122S-3126S
references: 11110885
references: Eur J Pharmacol. 2010 Dec 15;649(1-3):398-402
references: 20869961
references: Chin J Integr Med. 2012 Apr;18(4):293-8
references: 22457141
references: J Bone Miner Metab. 2003;21(5):268-75
references: 12928827
references: Phytomedicine. 2012 Oct 15;19(13):1147-55
references: 22951392
references: Science. 1999 Apr 2;284(5411):143-7
references: 10102814
references: Cochrane Database Syst Rev. 2006 Jul 19;(3):CD004196
references: 16856037
references: Bone. 1999 Aug;25(2):211-5
references: 10456387
references: Bone. 2012 Feb;50(2):477-89
references: 21872687
references: Genes Dev. 2003 Jun 1;17(11):1352-65
references: 12782654
references: Genes Dev. 1997 Dec 15;11(24):3286-305
references: 9407023
references: Steroids. 2010 Aug-Sep;75(8-9):565-9
references: 19778547
references: J Clin Invest. 2005 Dec;115(12):3318-25
references: 16322775
references: Oncogene. 1999 Dec 20;18(55):7860-72
references: 10630639
references: Rev Endocr Metab Disord. 2006 Jun;7(1-2):33-9
references: 16960757
references: J Osteoporos. 2011;2011:293808
references: 21876833
references: Chin J Integr Med. 2012 Apr;18(4):276-82
references: 22457138
references: Arch Intern Med. 2001 May 14;161(9):1161-72
references: 11343439
references: J Clin Pathol. 2002 Sep;55(9):693-8
references: 12195001
references: Proc Natl Acad Sci U S A. 2005 Apr 26;102(17):6207-12
references: 15833818
references: Bone. 1998 Mar;22(3):201-9
references: 9514212
references: J Biol Chem. 2002 Aug 23;277(34):30998-1004
references: 12055200
references: Alcohol Clin Exp Res. 1996 Apr;20(2):221-7
references: 8730211