Skip to main content
Top
Published in: BMC Complementary Medicine and Therapies 1/2017

Open Access 01-12-2017 | Research article

Sophorae Flos extract inhibits RANKL-induced osteoclast differentiation by suppressing the NF-κB/NFATc1 pathway in mouse bone marrow cells

Authors: Jeong-Mi Kim, Jung-Han Lee, Guem-San Lee, Eun-mi Noh, Hyun-Kyung Song, Dong Ryun Gu, Seong-Cheol Kim, Seoung Hoon Lee, Kang-Beom Kwon, Young-Rae Lee

Published in: BMC Complementary Medicine and Therapies | Issue 1/2017

Login to get access

Abstract

Background

Sophorae Flos (SF) is a composite of flowers and buds of Styphnolobium japonicum (L.) Schott and has been used in traditional Korean and Chinese medicine for the treatment of hemostasis and inflammation. Previous studies reported that SF possesses anti-obesity properties, as well as anti-allergic, anti-proliferative, and anti-inflammatory activities. However, the effect of SF in bone resorption has not been studies. In this study, we examined the potential of SF extract (SFE) to inhibit receptor activator of NF-κB ligand (RANKL) -induced osteoclast differentiation in cultured mouse-derived bone marrow macrophages (BMMs).

Methods

BMMs, that act as osteoclast precursors, were cultured with M-CSF (50 ng/ml) and RANKL (100 ng/ml) for 4 days to generate osteoclasts. Osteoclast differentiation was measured by tartrate-resistant acidic phosphatase (TRAP) staining and the TRAP solution assay. Osteoclast differentiation marker genes were analyzed by the quantitative real-time polymerase chain reaction analysis. RANKLs signaling pathways were confirmed through western blotting.

Results

SFE significantly decreased osteoclast differentiation in a dose-dependent manner. SFE inhibited RANKL-induced osteoclastogenesis by suppressing NF-κB activation. By contrast, SFE did not affect phospholipase C gamma 2 or subsequent cAMP response element binding activation. SFE inhibited the RANKL-induced expression of nuclear factor of activated T cells c1 (NFATc1).

Conclusions

SFE attenuated the RANKL-mediated induction of NF-κB through inhibition of IκBα phosphorylation, which contributed to inhibiting of RANKL-induced osteoclast differentiation through downregulation of NFATc1.
Literature
1.
go back to reference Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423:337–42.CrossRefPubMed Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423:337–42.CrossRefPubMed
2.
3.
go back to reference Akiyama T, Dass CR, Choong PF. Novel therapeutic strategy for osteosarcoma targeting osteoclast differentiation, bone-resorbing activity, and apoptosis pathway. Mol Cancer Ther. 2008;7:3461–9.CrossRefPubMed Akiyama T, Dass CR, Choong PF. Novel therapeutic strategy for osteosarcoma targeting osteoclast differentiation, bone-resorbing activity, and apoptosis pathway. Mol Cancer Ther. 2008;7:3461–9.CrossRefPubMed
5.
go back to reference Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, Morony S, Oliveira-dos-Santos AJ, Van G, Itie A, Khoo W, Wakeham A, Dunstan CR, Lacey DL, Mak TW, Boyle WJ, Penninger JM. OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature. 1999;397:315–23.CrossRefPubMed Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, Morony S, Oliveira-dos-Santos AJ, Van G, Itie A, Khoo W, Wakeham A, Dunstan CR, Lacey DL, Mak TW, Boyle WJ, Penninger JM. OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature. 1999;397:315–23.CrossRefPubMed
6.
go back to reference Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A, Tsuda E, Morinaga T, Higashio K, Udagawa N, Takahashi N, Suda T. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci U S A. 1998;95:3597–602.CrossRefPubMedPubMedCentral Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A, Tsuda E, Morinaga T, Higashio K, Udagawa N, Takahashi N, Suda T. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci U S A. 1998;95:3597–602.CrossRefPubMedPubMedCentral
7.
go back to reference Darnay BG, Haridas V, Ni J, Moore PA, Aggarwal BB. Characterization of the intracellular domain of receptor activator of NF-kappaB (RANK). Interaction with tumor necrosis factor receptor-associated factors and activation of NF-kappab and c-Jun N-terminal kinase. J Biol Chem. 1998;273:20551–5.CrossRefPubMed Darnay BG, Haridas V, Ni J, Moore PA, Aggarwal BB. Characterization of the intracellular domain of receptor activator of NF-kappaB (RANK). Interaction with tumor necrosis factor receptor-associated factors and activation of NF-kappab and c-Jun N-terminal kinase. J Biol Chem. 1998;273:20551–5.CrossRefPubMed
8.
go back to reference Lee SY, Reichlin A, Santana A, Sokol KA, Nussenzweig MC, Choi Y. TRAF2 is essential for JNK but not NF-kappaB activation and regulates lymphocyte proliferation and survival. Immunity. 1997;7:703–13.CrossRefPubMed Lee SY, Reichlin A, Santana A, Sokol KA, Nussenzweig MC, Choi Y. TRAF2 is essential for JNK but not NF-kappaB activation and regulates lymphocyte proliferation and survival. Immunity. 1997;7:703–13.CrossRefPubMed
9.
go back to reference Lee ZH, Kim HH. Signal transduction by receptor activator of nuclear factor kappa B in osteoclasts. Biochem Biophys Res Commun. 2003;305:211–4.CrossRefPubMed Lee ZH, Kim HH. Signal transduction by receptor activator of nuclear factor kappa B in osteoclasts. Biochem Biophys Res Commun. 2003;305:211–4.CrossRefPubMed
10.
go back to reference Anderson DM, Maraskovsky E, Billingsley WL, Dougall WC, Tometsko ME, Roux ER, Teepe MC, DuBose RF, Cosman D, Galibert L. A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature. 1997;390:175–9.CrossRefPubMed Anderson DM, Maraskovsky E, Billingsley WL, Dougall WC, Tometsko ME, Roux ER, Teepe MC, DuBose RF, Cosman D, Galibert L. A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature. 1997;390:175–9.CrossRefPubMed
11.
go back to reference 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. Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. Dev Cell. 2002;3:889–901.CrossRefPubMed 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. Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. Dev Cell. 2002;3:889–901.CrossRefPubMed
12.
go back to reference Ishida H, Umino T, Tsuji K, Kosuge T. Studies on the antihemostatic substances in herbs classified as hemostatics in traditional Chinese medicine. I. On the antihemostatic principles in Sophora japonica L. Chem Pharm Bull (Tokyo). 1989;37:1616–8.CrossRef Ishida H, Umino T, Tsuji K, Kosuge T. Studies on the antihemostatic substances in herbs classified as hemostatics in traditional Chinese medicine. I. On the antihemostatic principles in Sophora japonica L. Chem Pharm Bull (Tokyo). 1989;37:1616–8.CrossRef
13.
go back to reference Tang YP, Li YF, Hu J, Lou FC. Isolation and identification of antioxidants from Sophora japonica. J Asian Nat Prod Res. 2002;4:123–8.CrossRefPubMed Tang YP, Li YF, Hu J, Lou FC. Isolation and identification of antioxidants from Sophora japonica. J Asian Nat Prod Res. 2002;4:123–8.CrossRefPubMed
14.
go back to reference Liu R, Qi Y, Sun A, Xie H. Isolation and purification of chemical constituents from the pericarp of Sophora japonica L. by chromatography on a 12% cross-linked agarose gel. J Sep Sci. 2007;30:1870–4.CrossRefPubMed Liu R, Qi Y, Sun A, Xie H. Isolation and purification of chemical constituents from the pericarp of Sophora japonica L. by chromatography on a 12% cross-linked agarose gel. J Sep Sci. 2007;30:1870–4.CrossRefPubMed
15.
go back to reference Abe M, Suzuki O, Tasaki K, Tominaga K, Wakasa H. Analysis of lectin binding properties on human Burkitt’s lymphoma cell lines that show high spontaneous metastasis to distant organs in SCID mice: the binding sites for soybean agglutinin lectin masked by sialylation are closely associated with metastatic lymphoma cells. Pathol Int. 1996;46:977–83.CrossRefPubMed Abe M, Suzuki O, Tasaki K, Tominaga K, Wakasa H. Analysis of lectin binding properties on human Burkitt’s lymphoma cell lines that show high spontaneous metastasis to distant organs in SCID mice: the binding sites for soybean agglutinin lectin masked by sialylation are closely associated with metastatic lymphoma cells. Pathol Int. 1996;46:977–83.CrossRefPubMed
16.
go back to reference Kim BH, Chung EY, Min BK, Lee SH, Kim MK, Min KR, Kim Y. Anti-inflammatory action of legume isoflavonoid sophoricoside through inhibition on cyclooxygenase-2 activity. Planta Med. 2003;69:474–6.CrossRefPubMed Kim BH, Chung EY, Min BK, Lee SH, Kim MK, Min KR, Kim Y. Anti-inflammatory action of legume isoflavonoid sophoricoside through inhibition on cyclooxygenase-2 activity. Planta Med. 2003;69:474–6.CrossRefPubMed
17.
go back to reference Kim JM, Yun-Choi HS. Anti-platelet effects of flavonoids and flavonoid-glycosides from Sophora japonica. Arch Pharm Res. 2008;31:886–90.CrossRefPubMed Kim JM, Yun-Choi HS. Anti-platelet effects of flavonoids and flavonoid-glycosides from Sophora japonica. Arch Pharm Res. 2008;31:886–90.CrossRefPubMed
18.
go back to reference Park KW, Lee JE, Park KM. Diets containing Sophora japonica L. prevent weight gain in high-fat diet-induced obese mice. Nutr Res. 2009;29:819–24.CrossRefPubMed Park KW, Lee JE, Park KM. Diets containing Sophora japonica L. prevent weight gain in high-fat diet-induced obese mice. Nutr Res. 2009;29:819–24.CrossRefPubMed
19.
go back to reference Walsh MC, Kim N, Kadono Y, Rho J, Lee SY, Lorenzo J, Choi Y. Osteoimmunology: interplay between the immune system and bone metabolism. Annu Rev Immunol. 2006;24:33–63.CrossRefPubMed Walsh MC, Kim N, Kadono Y, Rho J, Lee SY, Lorenzo J, Choi Y. Osteoimmunology: interplay between the immune system and bone metabolism. Annu Rev Immunol. 2006;24:33–63.CrossRefPubMed
21.
go back to reference Yoon HJ, Seo CR, Kim M, Kim YJ, Song NJ, Jang WS, Kim BJ, Lee J, Hong JW, Nho CW, Park KW. Dichloromethane extracts of Sophora japonica L. stimulate osteoblast differentiation in mesenchymal stem cells. Nutr Res. 2013;33:1053–62.CrossRefPubMed Yoon HJ, Seo CR, Kim M, Kim YJ, Song NJ, Jang WS, Kim BJ, Lee J, Hong JW, Nho CW, Park KW. Dichloromethane extracts of Sophora japonica L. stimulate osteoblast differentiation in mesenchymal stem cells. Nutr Res. 2013;33:1053–62.CrossRefPubMed
22.
go back to reference Lee J, Kim KW, Kim HK, Chae SW, Jung JC, Kwon SH, Rheu CH. The effect of Rexflavone (Sophorae fructus extract) on menopausal symptoms in postmenopausal women: a randomized double-blind placebo controlled clinical trial. Arch Pharm Res. 2010;33:523–30.CrossRefPubMed Lee J, Kim KW, Kim HK, Chae SW, Jung JC, Kwon SH, Rheu CH. The effect of Rexflavone (Sophorae fructus extract) on menopausal symptoms in postmenopausal women: a randomized double-blind placebo controlled clinical trial. Arch Pharm Res. 2010;33:523–30.CrossRefPubMed
23.
go back to reference Lee SH, Kim T, Jeong D, Kim N, Choi Y. The tec family tyrosine kinase Btk Regulates RANKL-induced osteoclast maturation. J Biol Chem. 2008;283:11526–34.CrossRefPubMedPubMedCentral Lee SH, Kim T, Jeong D, Kim N, Choi Y. The tec family tyrosine kinase Btk Regulates RANKL-induced osteoclast maturation. J Biol Chem. 2008;283:11526–34.CrossRefPubMedPubMedCentral
24.
go back to reference Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.CrossRefPubMed Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.CrossRefPubMed
25.
go back to reference Wada T, Nakashima T, Hiroshi N, Penninger JM. RANKL-RANK signaling in osteoclastogenesis and bone disease. Trends Mol Med. 2006;12:17–25.CrossRefPubMed Wada T, Nakashima T, Hiroshi N, Penninger JM. RANKL-RANK signaling in osteoclastogenesis and bone disease. Trends Mol Med. 2006;12:17–25.CrossRefPubMed
26.
go back to reference Kite GC, Veitch NC, Boalch ME, Lewis GP, Leon CJ, Simmonds MS. Flavonol tetraglycosides from fruits of Styphnolobium japonicum (Leguminosae) and the authentication of Fructus Sophorae and Flos Sophorae. Phytochemistry. 2009;70:785–94.CrossRefPubMed Kite GC, Veitch NC, Boalch ME, Lewis GP, Leon CJ, Simmonds MS. Flavonol tetraglycosides from fruits of Styphnolobium japonicum (Leguminosae) and the authentication of Fructus Sophorae and Flos Sophorae. Phytochemistry. 2009;70:785–94.CrossRefPubMed
27.
go back to reference Sims NA, Walsh NC. Intercellular cross-talk among bone cells: new factors and pathways. Curr Osteoporos Rep. 2012;10:109–17.CrossRefPubMed Sims NA, Walsh NC. Intercellular cross-talk among bone cells: new factors and pathways. Curr Osteoporos Rep. 2012;10:109–17.CrossRefPubMed
28.
go back to reference Gohda J, Akiyama T, Koga T, Takayanagi H, Tanaka S, Inoue J. RANK-mediated amplification of TRAF6 signaling leads to NFATc1 induction during osteoclastogenesis. EMBO J. 2005;24:790–9.CrossRefPubMedPubMedCentral Gohda J, Akiyama T, Koga T, Takayanagi H, Tanaka S, Inoue J. RANK-mediated amplification of TRAF6 signaling leads to NFATc1 induction during osteoclastogenesis. EMBO J. 2005;24:790–9.CrossRefPubMedPubMedCentral
30.
go back to reference Boyce BF, Xing L, Franzoso G, Siebenlist U. Required and nonessential functions of nuclear factor-kappa B in bone cells. Bone. 1999;25:137–9.CrossRefPubMed Boyce BF, Xing L, Franzoso G, Siebenlist U. Required and nonessential functions of nuclear factor-kappa B in bone cells. Bone. 1999;25:137–9.CrossRefPubMed
31.
go back to reference Franzoso G, Carlson L, Xing L, Poljak L, Shores EW, Brown KD, Leonardi A, Tran T, Boyce BF, Siebenlist U. Requirement for NF-kappaB in osteoclast and B-cell development. Genes Dev. 1997;11:3482–96.CrossRefPubMedPubMedCentral Franzoso G, Carlson L, Xing L, Poljak L, Shores EW, Brown KD, Leonardi A, Tran T, Boyce BF, Siebenlist U. Requirement for NF-kappaB in osteoclast and B-cell development. Genes Dev. 1997;11:3482–96.CrossRefPubMedPubMedCentral
32.
go back to reference Takatsuna H, Asagiri M, Kubota T, Oka K, Osada T, Sugiyama C, Saito H, Aoki K, Ohya K, Takayanagi H, Umezawa K. Inhibition of RANKL-induced osteoclastogenesis by (-)-DHMEQ, a novel NF-kappaB inhibitor. J Bone Miner Res. 2005;20:653–62.CrossRefPubMed Takatsuna H, Asagiri M, Kubota T, Oka K, Osada T, Sugiyama C, Saito H, Aoki K, Ohya K, Takayanagi H, Umezawa K. Inhibition of RANKL-induced osteoclastogenesis by (-)-DHMEQ, a novel NF-kappaB inhibitor. J Bone Miner Res. 2005;20:653–62.CrossRefPubMed
33.
go back to reference Asagiri M, Sato K, Usami T, Ochi S, Nishina H, Yoshida H, Morita I, Wagner EF, Mak TW, Serfling E, Takayanagi H. Autoamplification of NFATc1 expression determines its essential role in bone homeostasis. J Exp Med. 2005;202:1261–9.CrossRefPubMedPubMedCentral Asagiri M, Sato K, Usami T, Ochi S, Nishina H, Yoshida H, Morita I, Wagner EF, Mak TW, Serfling E, Takayanagi H. Autoamplification of NFATc1 expression determines its essential role in bone homeostasis. J Exp Med. 2005;202:1261–9.CrossRefPubMedPubMedCentral
34.
go back to reference Kim K, Lee SH, Ha Kim J, Choi Y, Kim N. NFATc1 induces osteoclast fusion via up-regulation of Atp6v0d2 and the dendritic cell-specific transmembrane protein (DC-STAMP). Mol Endocrinol. 2008;22:176–85.CrossRefPubMed Kim K, Lee SH, Ha Kim J, Choi Y, Kim N. NFATc1 induces osteoclast fusion via up-regulation of Atp6v0d2 and the dendritic cell-specific transmembrane protein (DC-STAMP). Mol Endocrinol. 2008;22:176–85.CrossRefPubMed
36.
go back to reference Koga T, Inui M, Inoue K, Kim S, Suematsu A, Kobayashi E, Iwata T, Ohnishi H, Matozaki T, Kodama T, Taniguchi T, Takayanagi H, Takai T. Costimulatory signals mediated by the ITAM motif cooperate with RANKL for bone homeostasis. Nature. 2004;428:758–63.CrossRefPubMed Koga T, Inui M, Inoue K, Kim S, Suematsu A, Kobayashi E, Iwata T, Ohnishi H, Matozaki T, Kodama T, Taniguchi T, Takayanagi H, Takai T. Costimulatory signals mediated by the ITAM motif cooperate with RANKL for bone homeostasis. Nature. 2004;428:758–63.CrossRefPubMed
37.
go back to reference Sato K, Suematsu A, Nakashima T, Takemoto-Kimura S, Aoki K, Morishita Y, Asahara H, Ohya K, Yamaguchi A, Takai T, Kodama T, Chatila TA, Bito H, Takayanagi H. Regulation of osteoclast differentiation and function by the CaMK-CREB pathway. Nat Med. 2006;12:1410–6.CrossRefPubMed Sato K, Suematsu A, Nakashima T, Takemoto-Kimura S, Aoki K, Morishita Y, Asahara H, Ohya K, Yamaguchi A, Takai T, Kodama T, Chatila TA, Bito H, Takayanagi H. Regulation of osteoclast differentiation and function by the CaMK-CREB pathway. Nat Med. 2006;12:1410–6.CrossRefPubMed
Metadata
Title
Sophorae Flos extract inhibits RANKL-induced osteoclast differentiation by suppressing the NF-κB/NFATc1 pathway in mouse bone marrow cells
Authors
Jeong-Mi Kim
Jung-Han Lee
Guem-San Lee
Eun-mi Noh
Hyun-Kyung Song
Dong Ryun Gu
Seong-Cheol Kim
Seoung Hoon Lee
Kang-Beom Kwon
Young-Rae Lee
Publication date
01-12-2017
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2017
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-016-1550-x

Other articles of this Issue 1/2017

BMC Complementary Medicine and Therapies 1/2017 Go to the issue