Regulation of osteoclastogenesis by Simon extracts composed of caffeic acid and related compounds: successful suppression of bone destruction accompanied with adjuvant-induced arthritis in rats


Simon extracts are vitamin K1-rich food materials extracted from the leaves of the Simon sweet potato. Although vitamin K is known to stimulate bone formation, we postulated that Simon extracts also contain unknown biological compounds having the ability to regulate bone resorption. Here we prepared the vitamin K-free fraction from the Simon extracts and investigated the ability of this fraction on the differentiation of osteoclasts. A remarkable inhibitory effect of osteoclastogenesis was observed when osteoclast precursors were treated with this fraction in rat bone marrow culture systems as well as in a pure differentiation system using murine osteoclast precursor cell line. The vitamin K-free Simon extracts markedly suppressed severe bone destruction mediated by abundant osteoclasts associated with adjuvant-induced arthritis in rats. High performance liquid chromatography (HPLC) analysis revealed that the vitamin K-free Simon extracts contained three types of low molecular weight inhibitors for osteoclastogenesis; caffeic acid, chlorogenic acids and isochlorogenic acids. Among these substances, caffeic acid showed the most powerful inhibitory effects on osteoclastogenesis. Caffeic acid significantly suppressed expression of NFATc1, a key transcription factor for the induction of osteoclastogenesis. Our current study enlightened a high utility of the Simon extracts and their chemical components as effective regulators for bone resorption accompanied with inflammation and metabolic bone diseases.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Abramson SB, Amin A (2002) Blocking the effects of IL-1 in rheumatoid arthritis protects bone and cartilage. Rheumatology 41:972–980

  2. Andreakos ET, Foxwell BM, Brennan FM, Maini RN, Feldman M (2002) Cytokines and anti-cytokine biologicals in autoimmunity: present and future. Cytokine Growth Factor Rev 13:299–313

  3. Borrelli F, Maffia P, Pinto L, Ianaro A, Russo A, Capasso F, Ialenti A (2002) Phytochemical compounds involved in the anti-inflammatory effect of propolis extract. Fitoterapia 73(suppl. 1):S53–S63

  4. Chen YJ, Shiao MS, Wang SY (2001) The antioxidant caffeic acid phenethyl ester induces apoptosis associated with selective scavenging of hydrogen peroxide in human leukemic HL-60 cell. Anticancer Drugs 12:143–149

  5. Cimaz R (2002) Osteoporosis in childhood rheumatic diseases: prevention and therapy. Best Pract Res Clin Rheumatol 16:397–409

  6. Fitzpatrick LR, Wang J, Le T (2001) Caffeic acid phenethyl ester, an inhibitor of nuclear factor-κB, attenuates bacterial peptidoglycan polysaccharide-induced colitis in rats. J Pharmacol Exp Ther 299:915–920

  7. Gonthier MP, Verny MA, Besson C, Remesy C, Scalbert A (2003) Chlorogenic acid bioavailability largely depends on its metabolism by the gut microflora in rats. J Nutr 133:1853–1859

  8. Grigoriadis AE, Wang ZQ, Cecchini MG, Hofstetter W, Felix R, Fleisch HA, Wagner EF (1994) c-Fos: a key regulator of osteoclast-macrophage lineage determination and bone remodeling. Science 266:443–448

  9. Heer M (2002) Nutritional interventions related to bone turnover in European space mission and simulation models. J Nutr 18:853–856

  10. Hirayama T, Danks L, Sabokbar A, Athanasou NA (2002) Osteoclast formation and activity in the pathogenesis of osteoporosis in rheumatoid arthritis. Rheumatology 41:1232–1239

  11. Ikeda F, Nishimura R, Matsubara T, Tanaka S, Inoue J, Reddy SV, Hata K, Yamashita K, Hiraga T, Watanabe T, Kukita T, Yoshioka K, Rao A, Yoneda T (2004) Critical roles of c-Jun signaling in regulation of NFAT family and RANKL-regulated osteoclast differentiation. J Clin Invest 114:475–484

  12. Iotsova V, Caamano J, Loy J, Yang Y, Lewin A, Bravo R (1997) Osteopetrosis in mice lacking NF-kappaB1 and NF-kappaB2. Nat Med 3:1285–1289

  13. Ishida N, Hayashi K, Hoshijima M, Ogawa T, Koga S, Miyatake Y, Kumegawa M, Kimura T, Takeya T (2002) Large scale gene expression analysis of osteoclastogenesis in vitro and elucidation of NFAT2 as a key regulator. J Biol Chem 277:41147–41156

  14. Ishihara K, Hirano T (2002) IL-6 in autoimmune disease and chronic inflammatory proliferative disease. Cytokine Growth Factor Rev 13:357–368

  15. Jimi E, Nakamura I, Ikebe T, Akiyama S, Takahashi N, Suda T (1998) Activation of NF-κB is involved in the survival of osteoclasts promoted by interleukin-1. J Biol Chem 273:8799–8805

  16. Jue DM, Jeon KI, Jeong JY (1999) Nuclear factor κB (NF-κB) pathway as a therapeutic target in rheumatoid arthritis. J Korean Med Sci 14:231–238

  17. Khosla S and Kleerekoper M (1999) Biochemical markers of bone turnover. In: Favus MJ (ed) Primer on the metabolic bone diseases and disorders of mineral metabolism, 4th edn. Lippincott Williams & Wilkins, Philadelphia, pp 128–133

  18. Krol W, Scheller S, Czuba Z, Matsuno T, Zydowicz G, Shani J, Mos M (1996) Inhibition of neutrophils’ chemiluminescence by ethanol extract of propolis (EEP) and its phenolic components. J Ethnopharma 55:19–25

  19. Kukita A, Kukita T, Hata K, Kurisu K, Kohashi O (1993a) Heat-treated osteoblastic cell (ROS17/2.8)-conditioned medium induces the formation of osteoclast-like cells. Bone Miner 23:113–127

  20. Kukita A, Kukita T, Shin JH, Kohashi O (1993b) Induction of mononuclear precursor cells with osteoclastic phenotypes in a rat bone marrow culture system depleted of stromal cells. Biochem Biophys Res Commun 196:1383–1389

  21. Kukita A, Kukita T, Ouchida M, Maeda H, Yatsuki H, Kohashi O (1999) Osteoclast-derived zinc finger (OCZF) protein with POZ domain, a possible transcriptional repressor, is involved in osteoclastogenesis. Blood 94:1987–1997

  22. Kuratani T, Nagata K, Kukita T, Hotokebuchi T, Nakashima A, Iijima T (1998) Induction of abundant osteoclast-like multinucleated giant cells in adjuvant arthritic rats with accompanying disordered high bone turnover. Histol Histopathol 13:751–759

  23. Makarov SS (2001) NF-κB in rheumatoid arthritis: a pivotal regulator of inflammation, hyperplasia, and tissue destruction. Arthritis Res 3:200–206

  24. Moller HJ (1998) Connective tissue markers of rheumatoid arthritis. Scand J Clin Lab Invest 58:269–278

  25. Natarajan K, Singh S, Burke TR Jr, Grunberger D, Aggareal BB (1996) Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF-κB. Proc Natl Acad Sci USA 93: 9090–9095

  26. Orban Z, Mitsiades N, Burke TR Jr, Tsokos M, Chrousos GP (2000) Caffeic acid phenethyl ester induces leukocyte apoptosis, modulates nuclear factor-kappa B and suppresses acute inflammation. Neuroimmunomodulation 7:99–105

  27. Rodan GA, Martin TJ (2000) Therapeutic approaches to bone diseases. Science 289:1508–1514

  28. Romas E, Gillespie MT, Martin TJ (2002) Involvement of receptor activator of NF-κB ligand and tumor necrosis factor-α in bone destruction in rheumatoid arthritis. Bone 30:340–346

  29. Shankar VS, Pazianas M, Huang CL, Simon B, Adebanjo OA, Zaidi M (1995) Caffeine modulates Ca2+ receptor activation in isolated rat osteoclasts and induces intracellular Ca2+ release. Am J Physiol Renal Physiol 268: 447–454

  30. Tabb MM, Sun A, Zhou C, Grun F, Errandi J, Romero K, Pham H, Inoue S, Mallick S, Lin M (2003) Vitamin K2 regulation of bone homeostasis is mediated by the steroid and xenobiotic receptor SXR. J Biol Chem 278:43919–43927

  31. Takayanagi H, Kim S, Koga T, Nishina H, Isshiki M, Yoshida H, Saiura A, Isobe M, Yokochi T, Inoue J, Wagner EF, Mak TW, Kodama T, Taniguchi T (2002) Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. Dev Cell 3:889–901

  32. Teitelbaum SL, Ross FP (2003) Genetic regulation of osteoclast development and function. Nat Rev Genet 4:638–649

  33. Tezuka K, Tezuka Y, Maejima A, Sato T, Nemoto K, Kamioka H, Hakeda Y, Kumegawa M (1994) Molecular cloning of a possible cysteine proteinase predominantly express in osteoclasts. J Biol Chem 269:1106–1109

  34. Tondravi MM, McKercher SR, Anderson K, Erdmann JM, Quiroz M, Maki R, Teitelbaum SL (1997) Osteopetrosis in mice lacking haematopoietic transcription factor PU. 1. Nature 386:81–84

  35. Toraldo G, Roggia C, Qian WP, Pacifici R, Weitzmann M.N (2003) IL-7 induce bone loss in vivo by induction of receptor activator of nuclear factorκ B ligand and tumor necrosis factor αfrom T cells. Proc Natl Acad Sci USA 100:125–130

  36. Watanabe T, Kukita T, Kukita A, Wata N, Toh K, Nagata K, Nomiyama H, Iijima T (2004) Direct stimulation of osteoclastogenesis by MIP-1α: evidence obtained from studies using RAW 264 cell clone highly responsive to RANKL. J Endocrinol 180:193–201

  37. Zaidi M, Moonga BS, Adebanjo OA (1999) Novel mechanism of calcium handling by osteoclast: a review-hypothesis. Proc Assoc Am Physicians 111:319–327

  38. Zaidi M, Blair HC, Moonga BS, Abe E, Huang CL (2003) Osteoclastogenesis, bone resorption, and osteoclast-based therapeutics. J Bone Miner Res 18:599–609

  39. Zhao H P, Shuto T, Hirata G, Iwamoto Y (2000) Aminobisphosphonate (YM175) inhibits bone destruction in rat adjuvant arthritis. J Orthop Sci 5:397–403

Download references


We thank Dr. Go Hirata of Department of Orthopedic Surgery, Faculty of Medical Science, Kyushu University for kind suggestions. This work was supported in part by a Grant for Scientific Research from the Japanese Ministry of Education, Science, and Culture (project 14571738). The authors have no conflicting financial interests.

Author information

Correspondence to Toshio Kukita.

Additional information

Quan Yong Tang is on leave from Hebei Medical University of China

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tang, Q.Y., Kukita, T., Ushijima, Y. et al. Regulation of osteoclastogenesis by Simon extracts composed of caffeic acid and related compounds: successful suppression of bone destruction accompanied with adjuvant-induced arthritis in rats. Histochem Cell Biol 125, 215–225 (2006).

Download citation


  • Simon extracts
  • Osteoclastogenesis
  • Adjuvant-induced arthritis
  • Caffeic acid
  • Cathepsin K
  • Food materials
  • NFATc1