Inflammation Research

, Volume 68, Issue 5, pp 415–421 | Cite as

Fructose 1,6-bisphosphate inhibits osteoclastogenesis by attenuating RANKL-induced NF-κB/NFATc-1

  • L. Wilches-Buitrago
  • P. R. Viacava
  • F. Q. Cunha
  • J. C. Alves-Filho
  • S. Y. FukadaEmail author
Original Research Paper



Although some glycolytic intermediates have been shown to modulate several cell type formation and activation, the functional role of fructose 1,6-bisphosphate (FBP) on osteoclastogenesis is still unknown.


Osteoclastogenesis was evaluated on bone marrow preosteoclasts cultured with M-CSF − 30 ng/ml, RANKL − 10 ng/ml, and two concentrations of FBP (100 and 300 µM). TRAP-positive stained cells were counted, and osteoclastogenic marker genes expression were evaluated by qPCR. Osteoclasts resorption capacity was evaluated by the expression of specific enzymes and capacity to resorb a mineralized matrix. The NF-κB activation was detected using RAW 264.7, stably expressing luciferase on the NF-κB responsive promoter.


We show that FBP, the product of the first stage of glycolysis, inhibited RANKL-induced osteoclasts differentiation and TRAP activity. The treatment of preosteoclasts with FBP attenuated osteoclast fusion and formation, without affecting cell viability. Moreover, the inhibition of several osteoclastogenic marker genes expression (TRAP, OSCAR, DC-STAMP, Integrin αv, NFATc1) by FBP correlates with a reduction of mineralized matrix resorption capacity. The mechanism underlying FBP-inhibition of osteoclastogenesis involves NF-κB/NFATc1 signaling pathway inhibition.


Altogether these data show a protective role of a natural glycolytic intermediate in bone homeostasis that may have therapeutic benefit for osteolytic diseases.


Bone remodeling Osteoclasts NFATc1 Fructose 1,6-bisphosphate 



The author’s thanks Mayara Santos Gomes for her technical assistance. This study was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP grant No. 2015/09034-0; 2013/08216-2) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-130230/2016-2 scholarship to LWB).

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.


  1. 1.
    Alford AI, Kozloff KM, Hankenson KD. Extracellular matrix networks in bone remodeling. Int J Biochem Cell Biol. 2015;65:20–31.CrossRefPubMedGoogle Scholar
  2. 2.
    Adamopoulos IE, Mellins ED. Alternative pathways of osteoclastogenesis in inflammatory arthritis. Nat Rev Rheumatol. 2015;11:189–94.CrossRefPubMedGoogle Scholar
  3. 3.
    Leidig-Bruckner G, Ziegler R. Diabetes mellitus a risk for osteoporosis? Exp Clin Endocrinol Diabetes. 2001;109(Suppl 2):493–514.CrossRefGoogle Scholar
  4. 4.
    Kim JH, Kim N. Regulation of NFATc1 in osteoclast differentiation. Journal of Bone Metabolism. 2014;21:233–41.CrossRefPubMedGoogle Scholar
  5. 5.
    Takayanagi H, Kim S, Koga T, et al. Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. Dev Cell. 2002;3:889–901.CrossRefPubMedGoogle Scholar
  6. 6.
    Asagiri M, Takayanagi H. The molecular understanding of osteoclast differentiation. Bone. 2007;40:251–64.CrossRefPubMedGoogle Scholar
  7. 7.
    Kim JM, Jeong D, Kang HK, Jung SY, et al. Osteoclast precursors display dynamic metabolic shifts toward accelerated glucose metabolism at an early stage of RANKL-stimulated osteoclast differentiation. Cell Physiol Biochem. 2007;20:935–46.CrossRefPubMedGoogle Scholar
  8. 8.
    Lee YS, Kim YS, Lee SY, Kim GH, Kim BJ, Lee SH, et al. AMP-kinase acts as a negative regulator of RANKL in the differentiation of osteoclasts. Bone. 2010;47:926–37.CrossRefPubMedGoogle Scholar
  9. 9.
    Fong JE, Le Nihouannen D, Tiedemann K, Sadvakassova G, Barralet JE, Komarova SV. Moderate excess of pyruvate augments osteoclastogenesis. Biol Open. 2013;22(2(4):387–95.CrossRefGoogle Scholar
  10. 10.
    Williams JP, Blair HC, McDonald JM, et al. Regulation of osteoclastic bone resorption by glucose. Biochem Biophys Res Commun. 1997;235:646–51.CrossRefPubMedGoogle Scholar
  11. 11.
    Ahn H, Lee K, Kim JM, Kwon SH, Lee SH, Lee SY, Jeong D. Accelerated lactate dehydrogenase activity potentiates osteoclastogenesis via NFATc1 signaling. PLoS One. 2016;11:e0153886.CrossRefPubMedGoogle Scholar
  12. 12.
    Indo Y, Takeshita S, Ishii KA, Hoshii T, Aburatani H, Hirao A, Ikeda K. Metabolic regulation of osteoclast differentiation and function. J Bone Miner Res. 2013;28:2392–9.CrossRefPubMedGoogle Scholar
  13. 13.
    Jones W, Bianchi K. Aerobic glycolysis: beyond proliferation. Front Immunol. 2015;15:6:227.Google Scholar
  14. 14.
    Cohen JE, Atluri P, Taylor MD, Grand TJ, Liao GP, Panlilio CM, et al. Fructose 1,6-diphosphate administration attenuates post-ischemic ventricular dysfunction. Heart Lung Circ. 2006;15:119–23.CrossRefPubMedGoogle Scholar
  15. 15.
    Ahn SM, Hwang JS, Lee SH. Fructose 1,6-diphosphate alleviates UV-induced oxidative skin damage in hairless mice. Biol Pharm Bull. 2007;30:692–7.CrossRefPubMedGoogle Scholar
  16. 16.
    Xu K, Stringer JL. Pharmacokinetics of fructose-1,6-diphosphate after intraperitoneal and oral administration to adult rats. Pharmacol Res. 2008;57:234–8.CrossRefPubMedGoogle Scholar
  17. 17.
    Veras FP, Peres RS, Saraiva AL, Pinto LG, Louzada-Junior P, Cunha TM, et al. Fructose 1,6-bisphosphate, a high-energy intermediate of glycolysis, attenuates experimental arthritis by activating anti-inflammatory adenosinergic pathway. Scientific Reports. 2015;5:15171.CrossRefPubMedGoogle Scholar
  18. 18.
    Ma B, Zhang Q, Wu D, Wang YL, Hu YY, Cheng YP, Yang ZD, Zheng YY, Ying HJ. Strontium fructose 1,6-diphosphate prevents bone loss in a rat model of postmenopausal osteoporosis via the OPG/RANKL/RANK pathway. Acta Pharmacol Sin. 2012;33:479–89.CrossRefPubMedGoogle Scholar
  19. 19.
    Bordignon Nunes F, Meier Graziottin C, Alves Filho JC, Lunardelli A, Caberlon E, Peres A, Rodrigues De Oliveira J. Immunomodulatory effect of fructose-1,6-bisphosphate on T-lymphocytes. Int Immunopharmacol. 2003;3:267–72.CrossRefPubMedGoogle Scholar
  20. 20.
    Shinohara M, Takayanagi H. Analysis of NFATc1-centered transcription factor regulatory networks in osteoclast formation. Methods Mol Biol. 2014;1164:171–6.CrossRefPubMedGoogle Scholar
  21. 21.
    Sundaram K, Nishimura R, Senn J, Youssef RF, London SD, Reddy SV. RANK ligand signaling modulates the matrix metalloproteinase-9 gene expression during osteoclast differentiation. Exp Cell Res. 2007;313:168–78.CrossRefPubMedGoogle Scholar
  22. 22.
    Blair HC, Teitelbaum SL, Grosso LE, et al. Extracellular-matrix degradation at acid pH. Avian osteoclast acid collagenase isolation and characterization. Biochem J. 1993;290(Pt 3):873–84.CrossRefPubMedGoogle Scholar
  23. 23.
    Boyce BF, Yamashita T, Yao Z, Zhang Q, Li F, Xing L. Roles for NF-kappaB and c-Fos in osteoclasts. J Bone Miner Metab. 2005;23:11–15.Google Scholar
  24. 24.
    Yu M, Qi X, Moreno JL, Farber DL, Keegan AD. NF-κB signaling participates in both RANKL- and IL-4-induced macrophage fusion: receptor cross-talk leads to alterations in NF-κB pathways. J Immunol. 2011;187:1797–806.CrossRefPubMedGoogle Scholar

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© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Ribeirao Preto Medical School, Department of PharmacologyUniversity of Sao PauloRibeirao PretoBrazil
  2. 2.School of Pharmaceutical Sciences of Ribeirao Preto, Department of Physics and ChemistryUniversity of Sao PauloRibeirao PretoBrazil

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