Open Access

Zoledronic acid inhibits osteoclast differentiation and function through the regulation of NF-κB and JNK signalling pathways

  • Authors:
    • Xiao‑Lin Huang
    • Lie‑Yu Huang
    • Yu‑Ting Cheng
    • Fang Li
    • Qian Zhou
    • Chao Wu
    • Qian‑Hui Shi
    • Zhi‑Zhong Guan
    • Jian Liao
    • Wei Hong
  • View Affiliations

  • Published online on: May 23, 2019     https://doi.org/10.3892/ijmm.2019.4207
  • Pages: 582-592
  • Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

It is well known that extensive osteoclast formation plays a key role in osteoporosis in post‑menopausal women and the elderly. The suppression of extensive osteoclastogenesis and bone resorption may be an effective preventive strategy for osteoporosis. Zoledronic acid (ZOL) has been indicated to play an essential role in regulating bone mineral density and has already been used in large clinical trials. However, the effects of ZOL on osteoclastogenesis remain to be fully elucidated. Therefore, the present study aimed to determine the effects of ZOL on osteoclastogenesis, and to explore the corresponding signalling pathways. By using a cell viability assay, as well as in vitro osteoclastogenesis, immunofluorescence and resorption pit assays, we demonstrated that ZOL (0.1‑5 µM) suppressed receptor activator of nuclear factor‑κB ligand (RANKL)‑induced osteoclast differentiation and bone resorptive activity. Furthermore, western blot analysis and reverse transcription‑quantitative PCR indicated that ZOL inhibited the RANKL‑induced activation of NF‑κB and the phosphorylation of JNK in RAW264.7 cells, and subsequently decreased the expression of osteoclastogenesis‑associated genes, including calcitonin receptor, tartrate‑resistant acid phosphatase and dendritic cell‑specific transmembrane protein. ZOL inhibited osteoclast formation and resorption in vitro by specifically suppressing NF‑κB and JNK signalling. On the whole, the findings of this study indicate that ZOL may serve as a potential agent for the treatment of osteoclast‑associated diseases, including osteoporosis.

References

1 

Kim HS, Suh KS, Sul D, Kim BJ, Lee SK and Jung WW: The inhibitory effect and the molecular mechanism of glabridin on RANKL-induced osteoclastogenesis in RAW264.7 cells. Int J Mol Med. 29:169–177. 2012.

2 

Villa A, Guerrini MM, Cassani B, Pangrazio A and Sobacchi C: Infantile malignant, autosomal recessive osteopetrosis: The rich and the poor. Calcif Tissue Int. 84:1–12. 2009. View Article : Google Scholar

3 

Boyle WJ, Simonet WS and Lacey DL: Osteoclast differentiation and activation. Nature. 423:337–342. 2003. View Article : Google Scholar : PubMed/NCBI

4 

Teitelbaum SL: Bone resorption by osteoclasts. Science. 289:1504–1508. 2000. View Article : Google Scholar : PubMed/NCBI

5 

Nijweide PJ, Burger EH and Feyen JH: Cells of bone: Proliferation, differentiation, and hormonal regulation. Physiol Rev. 66:855–886. 1986. View Article : Google Scholar : PubMed/NCBI

6 

Soysa NS, Alles N, Aoki K and Ohya K: Osteoclast formation and differentiation: An overview. J Med Dent Sci. 59:65–74. 2012.PubMed/NCBI

7 

Zeng Z, Zhang C and Chen J: Lentivirus-mediated RNA interference of DC-STAMP expression inhibits the fusion and resorptive activity of human osteoclasts. J Bone Miner Metab. 31:409–416. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Zeng XZ, He LG, Wang S, Wang K, Zhang YY, Tao L, Li XJ and Liu SW: Aconine inhibits RANKL-induced osteoclast differentiation in RAW264.7 cells by suppressing NF-κB and NFATc1 activation and DC-STAMP expression. Acta Pharmacol Sin. 37:255–263. 2016. View Article : Google Scholar

9 

Oikawa T, Kuroda Y and Matsuo K: Regulation of osteoclasts by membrane-derived lipid mediators. Cell Mol Life Sci. 70:3341–3353. 2013. View Article : Google Scholar : PubMed/NCBI

10 

Liou YM, Chan CL, Huang R and Wang CA: Effect of l-caldesmon on osteoclastogenesis in RANKL-induced RAW264.7 cells. J Cell Physiol. 233:6888–6901. 2018. View Article : Google Scholar : PubMed/NCBI

11 

Islam R, Bae HS, Yoon WJ, Woo KM, Baek JH, Kim HH, Uchida T and Ryoo HM: Pin1 regulates osteoclast fusion through suppression of the master regulator of cell fusion DC-STAMP. J Cell Physiol. 229:2166–2174. 2014. View Article : Google Scholar : PubMed/NCBI

12 

Abe K, Yoshimura Y, Deyama Y, Kikuiri T, Hasegawa T, Tei K, Shinoda H, Suzuki K and Kitagawa Y: Effects of bisphosphonates on osteoclastogenesis in RAW264.7 cells. Int J Mol Med. 29:1007–1015. 2012.PubMed/NCBI

13 

Mediero A, Perez-Aso M and Cronstein BN: Activation of adenosine A(2A) receptor reduces osteoclast formation via PKA- and ERK1/2-mediated suppression of NFκB nuclear trans-location. Br J Pharmacol. 169:1372–1388. 2013. View Article : Google Scholar : PubMed/NCBI

14 

Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, Elliott R, Colombero A, Elliott G, Scully S, et al: Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 93:165–176. 1998. View Article : Google Scholar : PubMed/NCBI

15 

Yuan M, Chen J and Zeng Z: Knockdown of macrophage inhibitory cytokine-1 in RPMI-8226 human multiple myeloma cells inhibits osteoclastic differentiation through inhibiting the RANKL-Erk1/2 signaling pathway. Mol Med Rep. 14:5199–5204. 2016. View Article : Google Scholar : PubMed/NCBI

16 

Wada T, Nakashima T, Hiroshi N and Penninger JM: RANKL-RANK signaling in osteoclastogenesis and bone disease. Trends Mol Med. 12:17–25. 2006. View Article : Google Scholar

17 

Huang P, Han J and Hui L: MAPK signaling in inflammation- associated cancer development. Protein Cell. 1:218–226. 2010. View Article : Google Scholar

18 

Ihn HJ, Lee D, Lee T, Shin HI, Bae YC, Kim SH and Park EK: The 1,2,3-triazole derivative KP-A021 suppresses osteoclast differentiation and function by inhibiting RANKL-mediated MEK-ERK signaling pathway. Exp Biol Med (Maywood). 240:1690–1697. 2015. View Article : Google Scholar

19 

Negishi-Koga T and Takayanagi H: Ca2+-NFATc1 signaling is an essential axis of osteoclast differentiation. Immunol Rev. 231:241–256. 2009. View Article : Google Scholar : PubMed/NCBI

20 

Zhang Y, Wang Z, Xie X, Wang J, Wang Y, Peng QS, Zhang M, Wu D, Liu N, Wang HB and Sun WC: Tatarinan N inhibits osteoclast differentiation through attenuating NF-κB, MAPKs and Ca2+-dependent signaling. Int Immunopharmacol. 65:199–211. 2018. View Article : Google Scholar : PubMed/NCBI

21 

Nakashima T and Takayanagi H: Osteoimmunology: Crosstalk between the immune and bone systems. J Clin Immunol. 29:555–567. 2009. View Article : Google Scholar : PubMed/NCBI

22 

Lambrinoudaki I, Vlachou S, Galapi F, Papadimitriou D and Papadias K: Once-yearly zoledronic acid in the prevention of osteoporotic bone fractures in postmenopausal women. Clin Interv Aging. 3:445–451. 2008. View Article : Google Scholar : PubMed/NCBI

23 

Dalle Carbonare L, Zanatta M, Gasparetto A and Valenti MT: Safety and tolerability of zoledronic acid and other bisphosphonates in osteoporosis management. Drug Healthc Patient Saf. 2:121–137. 2010. View Article : Google Scholar : PubMed/NCBI

24 

Benford HL, McGowan NW, Helfrich MH, Nuttall ME and Rogers MJ: Visualization of bisphosphonate-induced caspase-3 activity in apoptotic osteoclasts in vitro. Bone. 28:465–473. 2001. View Article : Google Scholar : PubMed/NCBI

25 

Yasen M, Li X, Jiang L, Yuan W, Che W and Dong J: Effect of zoledronic acid on spinal fusion outcomes in an ovariectomized rat model of osteoporosis. J Orthop Res. 33:1297–1304. 2015. View Article : Google Scholar : PubMed/NCBI

26 

Kimachi K, Kajiya H, Nakayama S, Ikebe T and Okabe K: Zoledronic acid inhibits RANK expression and migration of osteoclast precursors during osteoclastogenesis. Naunyn Schmiedebergs Arch Pharmacol. 383:297–308. 2011. View Article : Google Scholar : PubMed/NCBI

27 

Tsubaki M, Komai M, Itoh T, Imano M, Sakamoto K, Shimaoka H, Takeda T, Ogawa N, Mashimo K, Fujiwara D, et al: Nitrogen-containing bisphosphonates inhibit RANKL- and M-CSF-induced osteoclast formation through the inhibition of ERK1/2 and Akt activation. J Biomed Sci. 21:102014. View Article : Google Scholar : PubMed/NCBI

28 

Tai TW, Su FC, Chen CY, Jou IM and Lin CF: Activation of p38 MAPK-regulated Bcl-xL signaling increases survival against zoledronic acid-induced apoptosis in osteoclast precursors. Bone. 67:166–174. 2014. View Article : Google Scholar : PubMed/NCBI

29 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar

30 

Teitelbaum SL: Osteoclasts: What do they do and how do they do it? Am J Pathol. 170:427–435. 2007. View Article : Google Scholar : PubMed/NCBI

31 

Jurdic P, Saltel F, Chabadel A and Destaing O: Podosome and sealing zone: Specificity of the osteoclast model. Eur J Cell Biol. 85:195–202. 2006. View Article : Google Scholar : PubMed/NCBI

32 

Stevenson DA, Schwarz EL, Carey JC, Viskochil DH, Hanson H, Bauer S, Weng HY, Greene T, Reinker K, Swensen J, et al: Bone resorption in syndromes of the Ras/MAPK pathway. Clin Genet. 80:566–573. 2011. View Article : Google Scholar : PubMed/NCBI

33 

Suda T, Kobayashi K, Jimi E, Udagawa N and Takahashi N: The molecular basis of osteoclast differentiation and activation. Novartis Found Symp. 232:235–247; discussion 247-250. 2001.PubMed/NCBI

34 

Li DZ, Zhang QX, Dong XX, Li HD and Ma X: Treatment with hydrogen molecules prevents RANKL-induced osteoclast differentiation associated with inhibition of ROS formation and inactivation of MAPK, AKT and NF-kappa B pathways in murine RAW264.7 cells. J Bone Miner Metab. 32:494–504. 2014. View Article : Google Scholar

35 

Ikeda F, Nishimura R, Matsubara T, Tanaka S, Inoue J, Reddy SV, Hata K, Yamashita K, Hiraga T, Watanabe T, et al: Critical roles of c-Jun signaling in regulation of NFAT family and RANKL-regulated osteoclast differentiation. J Clin Invest. 114:475–484. 2004. View Article : Google Scholar : PubMed/NCBI

36 

Ikeda F, Matsubara T, Tsurukai T, Hata K, Nishimura R and Yoneda T: JNK/c-Jun signaling mediates an anti-apoptotic effect of RANKL in osteoclasts. J Bone Miner Res. 23:907–914. 2008. View Article : Google Scholar : PubMed/NCBI

37 

Siddiqi MH, Siddiqi MZ, Kang S, Noh HY, Ahn S, Simu SY, Aziz MA, Sathishkumar N, Jiménez Pérez ZE and Yang DC: Inhibition of osteoclast differentiation by ginsenoside rg3 in RAW264.7 cells via RANKL, JNK and p38 MAPK pathways through a modulation of cathepsin k: An in silico and in vitro study. Phytother Res. 29:1286–1294. 2015. View Article : Google Scholar : PubMed/NCBI

38 

Feng X and McDonald JM: Disorders of bone remodeling. Annu Rev Pathol. 6:121–145. 2011. View Article : Google Scholar

39 

Wu K, Lin TH, Liou HC, Lu DH, Chen YR, Fu WM and Yang RS: Dextromethorphan inhibits osteoclast differentiation by suppressing RANKL-induced nuclear factor-κB activation. Osteoporos Int. 24:2201–2214. 2013. View Article : Google Scholar : PubMed/NCBI

40 

Kang MR, Jo SA, Yoon YD, Park KH, Oh SJ, Yun J, Lee CW, Nam KH, Kim Y, Han SB, et al: Agelasine D suppresses RANKL-induced osteoclastogenesis via down-regulation of c-Fos, NFATc1 and NF-κB. Mar Drugs. 12:5643–5656. 2014. View Article : Google Scholar : PubMed/NCBI

41 

Lee CC, Liu FL, Chen CL, Chen TC, Chang DM and Huang HS: Discovery of 5-(2′,4′-difluorophenyl)-salicylanilides as new inhibitors of receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis. Eur J Med Chem. 98:115–126. 2015. View Article : Google Scholar : PubMed/NCBI

42 

Liu W and Zhang X: Receptor activator of nuclear factor-κB ligand (RANKL)/RANK/osteoprotegerin system in bone and other tissues (review). Mol Med Rep. 11:3212–3218. 2015. View Article : Google Scholar : PubMed/NCBI

43 

Zhao XL, Chen JJ, Si SY, Chen LF and Wang Z: T63 inhibits osteoclast differentiation through regulating MAPKs and Akt signaling pathways. Eur J Pharmacol. 834:30–35. 2018. View Article : Google Scholar : PubMed/NCBI

44 

Kim HJ, Yoon KA, Lee MK, Kim SH, Lee IK and Kim SY: A novel small molecule, NecroX-7, inhibits osteoclast differentiation by suppressing NF-κB activity and c-Fos expression. Life Sci. 91:928–934. 2012. View Article : Google Scholar : PubMed/NCBI

45 

Leotoing L, Wauquier F, Guicheux J, Miot-Noirault E, Wittrant Y and Coxam V: The polyphenol fisetin protects bone by repressing NF-κB and MKP-1-dependent signaling pathways in osteoclasts. PLoS One. 8:e683882013. View Article : Google Scholar

46 

Kong X, Wu W, Yang Y, Wan H, Li X, Zhong M, Zhao H, Su X, Jia S, Ju D and Lin N: Total saponin from Anemone flaccida Fr. Schmidt abrogates osteoclast differentiation and bone resorption via the inhibition of RANKL-induced NF-κB, JNK and p38 MAPKs activation. J Transl Med. 13:912015. View Article : Google Scholar

47 

Xu X, Liu N, Wang Y, Pan LC, Wu D, Peng Q, Zhang M, Wang HB and Sun WC: Tatarinan O, a lignin-like compound from the roots of Acorus tatarinowii Schott inhibits osteoclast differentiation through suppressing the expression of c-Fos and NFATc1. Int Immunopharmacol. 34:212–219. 2016. View Article : Google Scholar : PubMed/NCBI

48 

Fliefel R, Troltzsch M, Kuhnisch J, Ehrenfeld M and Otto S: Treatment strategies and outcomes of bisphosphonate-related osteonecrosis of the jaw (BRONJ) with characterization of patients: A systematic review. Int J Oral Maxillofac Surg. 44:568–585. 2015. View Article : Google Scholar : PubMed/NCBI

49 

Sakaguchi O, Kokuryo S, Tsurushima H, Tanaka J, Habu M, Uehara M, Nishihara T and Tominaga K: Lipopolysaccharide aggravates bisphosphonate-induced osteonecrosis in rats. Int J Oral Maxillofac Surg. 44:528–534. 2015. View Article : Google Scholar

50 

Wachi T, Shuto T, Shinohara Y, Matono Y and Makihira S: Release of titanium ions from an implant surface and their effect on cytokine production related to alveolar bone resorption. Toxicology. 327:1–9. 2015. View Article : Google Scholar

51 

Baek JM, Kim JY, Lee CH, Yoon KH and Lee MS: Methyl gallate inhibits osteoclast formation and function by suppressing Akt and Btk-PLCgamma2-Ca(2+) signaling and prevents lipo-polysaccharide-induced bone loss. Int J Mol Sci. 18:E5812017. View Article : Google Scholar

52 

Sun X, Wei J, Lyu J, Bian T, Liu Z, Huang J, Pi F, Li C and Zhong Z: Bone-targeting drug delivery system of biomineral-binding liposomes loaded with icariin enhances the treatment for osteoporosis. J Nanobiotechnology. 17:102019. View Article : Google Scholar : PubMed/NCBI

53 

Yasuda H: The mechanism of anti-RANKL antibody in the treatment of metabolic bone diseases including osteoporosis-possible applications of anti-RANKL antibody to the treatment of cancer patients. Nihon Yakurigaku Zasshi. 153:11–15. 2019.In Japanese. View Article : Google Scholar

54 

Moen MD and Keam SJ: Denosumab: A review of its use in the treatment of postmenopausal osteoporosis. Drugs Aging. 28:63–82. 2011. View Article : Google Scholar

55 

Sidlauskas KM, Sutton EE and Biddle MA: Osteoporosis in men: Epidemiology and treatment with denosumab. Clin Interv Aging. 9:593–601. 2014.PubMed/NCBI

56 

Tai TW, Chen CY, Su FC, Tu YK, Tsai TT, Lin CF and Jou IM: Reactive oxygen species are required for zoledronic acid-induced apoptosis in osteoclast precursors and mature osteoclast-like cells. Sci Re. 7:442452017.

57 

Lyles KW, Colon-Emeric CS, Magaziner JS, Adachi JD, Pieper CF, Mautalen C, Hyldstrup L, Recknor C, Nordsletten L, Moore KA, et al: Zoledronic acid and clinical fractures and mortality after hip fracture. N Engl J Med. 357:1799–1809. 2007. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
APA
Huang, X., Huang, L., Cheng, Y., Li, F., Zhou, Q., Wu, C. ... Hong, W. (2019). Zoledronic acid inhibits osteoclast differentiation and function through the regulation of NF-κB and JNK signalling pathways. International Journal of Molecular Medicine, 44, 582-592. https://doi.org/10.3892/ijmm.2019.4207
MLA
Huang, X., Huang, L., Cheng, Y., Li, F., Zhou, Q., Wu, C., Shi, Q., Guan, Z., Liao, J., Hong, W."Zoledronic acid inhibits osteoclast differentiation and function through the regulation of NF-κB and JNK signalling pathways". International Journal of Molecular Medicine 44.2 (2019): 582-592.
Chicago
Huang, X., Huang, L., Cheng, Y., Li, F., Zhou, Q., Wu, C., Shi, Q., Guan, Z., Liao, J., Hong, W."Zoledronic acid inhibits osteoclast differentiation and function through the regulation of NF-κB and JNK signalling pathways". International Journal of Molecular Medicine 44, no. 2 (2019): 582-592. https://doi.org/10.3892/ijmm.2019.4207