Sirtuin 1 inhibits TNF-α-mediated osteoclastogenesis of bone marrow-derived macrophages through both ROS generation and TRPV1 activation
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Sirtuin 1 (SIRT1), also known as NAD-dependent deacetylase, has been reported to increase in vivo osteoclast-mediated bone resorption. However, its effects on osteoclastogenesis or bone loss in vitro have not been widely examined. Therefore, the effects and underlying mechanism of SIRT1 on osteoclast differentiation in mice in vitro were studied. During RANKL-induced osteoclastogenesis in differentiated bone marrow-derived macrophages (BMMs), SIRT1 downregulation was observed. The use of resveratrol (SIRT1 activator) and SIRT1 overexpression was found to inhibit osteoclastogenesis, which was confirmed by TRAP staining and activity loss, reduced expression of osteoclast markers and related genes, and a decrease in the number of multinuclear cells. In contrast, treatment with EX-527 (SIRT1 inhibitor) as well as SIRT1 silencing promoted osteoclastogenesis. Furthermore, the tumor necrosis factor (TNF)-α level was reduced by resveratrol treatment and SIRT1 overexpression but increased following EX-527 incubation and SIRT1 depletion. TNF-α silencing blocked the osteoclastogenesis of BMMs promoted by SIRT1 depletion. Moreover, transient receptor potential vanilloid 1 (TRPV1) channel activation and reactive oxygen species (ROS) production, which are associated with osteoclastogenesis, were impaired by TNF-α silencing. These data demonstrate that SIRT1 directly inhibits osteoclastogenesis by inhibiting ROS generation and TRPV1 channel activation under mediation of TNF-α.
KeywordsOsteoclastogenesis Bone marrow-derived macrophages Sirtuin 1 Tumor necrosis factor α Reactive oxygen species
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
- 6.McHugh KP, Hodivala-Dilke K, Zheng MH, Namba N, Lam J, Novack D, Feng X, Ross FP, Hynes RO, Teitelbaum SL (2000) Mice lacking beta3 integrins are osteosclerotic because of dysfunctional osteoclasts. J Clin Invest 105(4):433–440. https://doi.org/10.1172/JCI8905 CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Gravallese EM, Manning C, Tsay A, Naito A, Pan C, Amento E, Goldring SR (2000) Synovial tissue in rheumatoid arthritis is a source of osteoclast differentiation factor. Arthritis Rheum 43 (2):250–258. https://doi.org/10.1002/1529-0131(200002)43:2%3C250::Aid-anr3%3E3.0.Co;2-p CrossRefPubMedGoogle Scholar
- 11.Takayanagi H, Iizuka H, Juji T, Nakagawa T, Yamamoto A, Miyazaki T, Koshihara Y, Oda H, Nakamura K, Tanaka S (2000) Involvement of receptor activator of nuclear factor kappaB ligand/osteoclast differentiation factor in osteoclastogenesis from synoviocytes in rheumatoid arthritis. Arthritis Rheum 43 (2):259–269. https://doi.org/10.1002/1529-0131(200002)43:2%3C259::Aid-anr4%3E3.0.Co;2-w CrossRefPubMedGoogle Scholar
- 12.Harris WH (1995) The problem is osteolysis. Clin Orthop Relat Res 311:46–53Google Scholar
- 19.Cohen-Kfir E, Artsi H, Levin A, Abramowitz E, Bajayo A, Gurt I, Zhong L, D’Urso A, Toiber D, Mostoslavsky R, Dresner-Pollak R (2011) Sirt1 is a regulator of bone mass and a repressor of Sost encoding for sclerostin, a bone formation inhibitor. Endocrinology 152(12):4514–4524. https://doi.org/10.1210/en.2011-1128 CrossRefPubMedGoogle Scholar
- 20.Simic P, Zainabadi K, Bell E, Sykes DB, Saez B, Lotinun S, Baron R, Scadden D, Schipani E, Guarente L (2013) SIRT1 regulates differentiation of mesenchymal stem cells by deacetylating beta-catenin. EMBO Mol Med 5(3):430–440. https://doi.org/10.1002/emmm.201201606 CrossRefPubMedPubMedCentralGoogle Scholar
- 21.Iyer S, Han L, Bartell SM, Kim HN, Gubrij I, de Cabo R, O’Brien CA, Manolagas SC, Almeida M (2014) Sirtuin1 (Sirt1) promotes cortical bone formation by preventing beta-catenin sequestration by FoxO transcription factors in osteoblast progenitors. J Biol Chem 289(35):24069–24078. https://doi.org/10.1074/jbc.M114.561803 CrossRefPubMedPubMedCentralGoogle Scholar
- 22.Edwards JR, Perrien DS, Fleming N, Nyman JS, Ono K, Connelly L, Moore MM, Lwin ST, Yull FE, Mundy GR, Elefteriou F (2013) Silent information regulator (Sir)T1 inhibits NF-kappaB signaling to maintain normal skeletal remodeling. J Bone Miner Res 28(4):960–969. https://doi.org/10.1002/jbmr.1824 CrossRefPubMedGoogle Scholar
- 23.Gurt I, Artsi H, Cohen-Kfir E, Hamdani G, Ben-Shalom G, Feinstein B, El-Haj M, Dresner-Pollak R (2015) The Sirt1 activators SRT2183 and SRT3025 inhibit RANKL-induced osteoclastogenesis in bone marrow-derived macrophages and down-regulate Sirt3 in Sirt1 null cells. PLoS ONE 10(7):e0134391. https://doi.org/10.1371/journal.pone.0134391 CrossRefPubMedPubMedCentralGoogle Scholar
- 24.He X, Andersson G, Lindgren U, Li Y (2010) Resveratrol prevents RANKL-induced osteoclast differentiation of murine osteoclast progenitor RAW 264.7 cells through inhibition of ROS production. Biochem Biophys Res Commun 401(3):356–362. https://doi.org/10.1016/j.bbrc.2010.09.053 CrossRefPubMedGoogle Scholar
- 31.Shakibaei M, Buhrmann C, Mobasheri A (2011) Resveratrol-mediated SIRT-1 interactions with p300 modulate receptor activator of NF-kappaB ligand (RANKL) activation of NF-kappaB signaling and inhibit osteoclastogenesis in bone-derived cells. J Biol Chem 286(13):11492–11505. https://doi.org/10.1074/jbc.M110.198713 CrossRefPubMedPubMedCentralGoogle Scholar
- 32.Nakamura T, Imai Y, Matsumoto T, Sato S, Takeuchi K, Igarashi K, Harada Y, Azuma Y, Krust A, Yamamoto Y, Nishina H, Takeda S, Takayanagi H, Metzger D, Kanno J, Takaoka K, Martin TJ, Chambon P, Kato S (2007) Estrogen prevents bone loss via estrogen receptor alpha and induction of Fas ligand in osteoclasts. Cell 130(5):811–823. https://doi.org/10.1016/j.cell.2007.07.025 CrossRefGoogle Scholar
- 34.Gertz M, Fischer F, Nguyen GTT, Lakshminarasimhan M, Schutkowski M, Weyand M, Steegborn C (2013) Ex-527 inhibits Sirtuins by exploiting their unique NAD+-dependent deacetylation mechanism. Proc Natl Acad Sci USA 110(30):E2772–E2781. https://doi.org/10.1073/pnas.1303628110 CrossRefPubMedGoogle Scholar
- 36.Kim MS, Yang YM, Son A, Tian YS, Lee SI, Kang SW, Muallem S, Shin DM (2010) RANKL-mediated reactive oxygen species pathway that induces long lasting Ca2 + oscillations essential for osteoclastogenesis. J Biol Chem 285(10):6913–6921. https://doi.org/10.1074/jbc.M109.051557 CrossRefPubMedPubMedCentralGoogle Scholar
- 38.Wang Y, Gao Y, Tian Q, Deng Q, Wang Y, Zhou T, Liu Q, Mei K, Wang Y, Liu H, Ma R, Ding Y, Rong W, Cheng J, Yao J, Xu TL, Zhu MX, Li Y (2018) TRPV1 SUMOylation regulates nociceptive signaling in models of inflammatory pain. Nat Commun 9(1):1529. https://doi.org/10.1038/s41467-018-03974-7 CrossRefPubMedPubMedCentralGoogle Scholar