1,25-Dihydroxyvitamin D3 attenuates disease severity and induces synoviocyte apoptosis in a concentration-dependent manner in rats with adjuvant-induced arthritis by inactivating the NF-κB signaling pathway

  • Hua-Qiang Sun
  • Dong Yan
  • Qin-Nan Wang
  • Hong-Zheng Meng
  • Ye-Yong Zhang
  • Lu-Xu Yin
  • Xin-Feng YanEmail author
  • Shu-Feng LiEmail author
Original Article


An aggressive proliferation of synoviocytes is the hallmark of rheumatoid arthritis (RA). Emerging evidence shows that inhibiting the NF-κB signaling pathway with 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] may be a therapeutic approach for controlling inflammatory diseases. In this study, we demonstrated the protective effects of three different 1,25(OH)2D3 concentration on adjuvant-induced arthritis (AA) rats through the NF-κB signaling pathway and their pro-apoptotic roles in cultured adjuvant-induced arthritis synoviocytes (AIASs). AA rats were prepared by injecting complete Freund’s adjuvant and independently given daily intraperitoneal injection of 1,25(OH)2D3 at concentrations of 50, 100, and 300 ng/day/kg. Subsequently, AIASs were isolated from the inflamed joints of AA rats to test the effects of 1,25(OH)2D3 on AIASs in vitro. Intraperitoneal injection of 1,25-(OH)2D3 was found to induce a concentration- and time-dependent improvement in relieving the symptoms of AA. We found an increased paw withdrawal thermal latency (PWTL) in the affected paw of AA rats as the concentration of 1,25-(OH)2D3 increased. 1,25-(OH)2D3 treatment reduced levels of inflammatory factors in synovial tissues of AA rats. In the case of cultured AIASs, 1,25-(OH)2D3 was shown to inhibit cell proliferation and induce cell apoptosis in a concentration-dependent manner. Additionally, 1,25-(OH)2D3 inhibited the activation of the NF-κB signaling pathway. In conclusion, our study provides evidence emphasizing that 1,25(OH)2D3 has the potential to attenuate disease severity in RA potentially due to its contributory role in synoviocyte proliferation and apoptosis. The protective role of 1,25(OH)2D3 against RA depends on the NF-κB signaling pathway.


1,25(OH)2D3 NF-κB signaling pathway Rheumatoid arthritis Synoviocyte Adjuvant-induced arthritis 



This study was supported by Key Research and Development Program of Shandong Province (no. 2016GSF201232) and the Natural Science Foundation of Shandong Province (no. ZR2017LH028). We would like to give our sincere appreciation to the reviewers for their helpful comments on this article.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

The study was conducted with the approval of the Animal Ethics Committee of Shandong Provincial Qianfoshan Hospital, Shandong University. All experimental procedures were performed on laboratory animals in accordance with institutional guidelines for the care and use of laboratory animals.


  1. 1.
    Mellado M, Martinez-Munoz L, Cascio G, Lucas P, Pablos JL, Rodriguez-Frade JM (2015) T cell migration in rheumatoid arthritis. Front Immunol 6:384CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Burkhardt J, Blume M, Petit-Teixeira E, Hugo Teixeira V, Steiner A et al (2014) Cellular adhesion gene SELP is associated with rheumatoid arthritis and displays differential allelic expression. PLoS One 9:e103872CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Zerbini CA, Lomonte AB (2012) Tofacitinib for the treatment of rheumatoid arthritis. Expert Rev Clin Immunol 8:319–331CrossRefPubMedGoogle Scholar
  4. 4.
    Smolen JS, Aletaha D, Bijlsma JW, Breedveld FC, Boumpas D et al (2010) Treating rheumatoid arthritis to target: recommendations of an international task force. Ann Rheum Dis 69:631–637CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Smolen JS, Landewe R, Breedveld FC, Buch M, Burmester G et al (2014) EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs: 2013 update. Ann Rheum Dis 73:492–509CrossRefPubMedGoogle Scholar
  6. 6.
    Smolen JS, Breedveld FC, Burmester GR, Bykerk V, Dougados M et al (2016) Treating rheumatoid arthritis to target: 2014 update of the recommendations of an international task force. Ann Rheum Dis 75:3–15CrossRefPubMedGoogle Scholar
  7. 7.
    Feely MG (2010) New and emerging therapies for the treatment of rheumatoid arthritis. Open Access Rheumatol 2:35–43CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Park SY, Lee SW, Shin HK, Chung WT, Lee WS, Rhim BY, Hong KW, Kim CD (2010) Cilostazol enhances apoptosis of synovial cells from rheumatoid arthritis patients with inhibition of cytokine formation via Nrf2-linked heme oxygenase 1 induction. Arthritis Rheum 62:732–741CrossRefPubMedGoogle Scholar
  9. 9.
    Noh EM, Kim JS, Hur H, Park BH, Song EK, Han MK, Kwon KB, Yoo WH, Shim IK, Lee SJ, Youn HJ, Lee YR (2009) Cordycepin inhibits IL-1beta-induced MMP-1 and MMP-3 expression in rheumatoid arthritis synovial fibroblasts. Rheumatology (Oxford) 48:45–48CrossRefGoogle Scholar
  10. 10.
    Susick L, Senanayake T, Veluthakal R, Woster PM, Kowluru A (2009) A novel histone deacetylase inhibitor prevents IL-1beta induced metabolic dysfunction in pancreatic beta-cells. J Cell Mol Med 13:1877–1885CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Yin G, Wang Y, Cen XM, Yang M, Liang Y, Xie QB (2015) Lipid peroxidation-mediated inflammation promotes cell apoptosis through activation of NF-kappaB pathway in rheumatoid arthritis synovial cells. Mediat Inflamm 2015:460310Google Scholar
  12. 12.
    Gonzalez-Pardo V, D’Elia N, Verstuyf A, Boland R, Russo de Boland A (2012) NFkappaB pathway is down-regulated by 1alpha,25(OH)(2)-vitamin D(3) in endothelial cells transformed by Kaposi sarcoma-associated herpes virus G protein coupled receptor. Steroids 77:1025–1032CrossRefPubMedGoogle Scholar
  13. 13.
    He XJ, Ding Y, Xiang W, Dang XQ (2016) Roles of 1,25(OH)2D3 and vitamin D receptor in the pathogenesis of rheumatoid arthritis and systemic lupus erythematosus by regulating the activation of CD4+ T cells and the PKCdelta/ERK signaling pathway. Cell Physiol Biochem 40:743–756CrossRefPubMedGoogle Scholar
  14. 14.
    Waksman BH (2002) Immune regulation in adjuvant disease and other arthritis models: relevance to pathogenesis of chronic arthritis. Scand J Immunol 56:12–34CrossRefPubMedGoogle Scholar
  15. 15.
    Boini S, Guillemin F (2001) Radiographic scoring methods as outcome measures in rheumatoid arthritis: properties and advantages. Ann Rheum Dis 60:817–827PubMedPubMedCentralGoogle Scholar
  16. 16.
    Wojciechowska M, Watroba M, Ciuzynska G, Bral M, Szukiewicz D (2013) Ischaemic heart preconditioning in rats with adjuvant-induced arthritis. Kardiol Pol 71:839–844CrossRefPubMedGoogle Scholar
  17. 17.
    Shah AB, DiMartino SJ, Trujillo G, Kew RR (2006) Selective inhibition of the C5a chemotactic cofactor function of the vitamin D binding protein by 1,25(OH)2 vitamin D3. Mol Immunol 43:1109–1115CrossRefPubMedGoogle Scholar
  18. 18.
    Kim TH, Lee B, Kwon E, Choi SJ, Lee YH, Song GG, Sohn J, Ji JD (2013) Regulation of TREM-1 expression by 1,25-dihydroxyvitamin D3 in human monocytes/macrophages. Immunol Lett 154:80–85CrossRefPubMedGoogle Scholar
  19. 19.
    Scanzello CR, Albert AS, DiCarlo E, Rajan KB, Kanda V, Asomugha EU, Swaim BH, Katz JN, Goldring SR, Richmond JC, McKeon B (2013) The influence of synovial inflammation and hyperplasia on symptomatic outcomes up to 2 years post-operatively in patients undergoing partial meniscectomy. Osteoarthritis Cartil 21:1392–1399CrossRefGoogle Scholar
  20. 20.
    Yurekli I, Gokalp O, Kiray M, Bademci M, Yetkin U, Ergunes K, Yilmaz O, Bayrak S, Gurbuz A (2013) Effect of 1alpha-25-dihydroxyvitamin D3 on intimal hyperplasia developing in vascular anastomoses: a rabbit model. Arch Med Sci 9:404–408CrossRefPubMedGoogle Scholar
  21. 21.
    van Hamburg JP, Asmawidjaja PS, Davelaar N, Mus AM, Cornelissen F, van Leeuwen JP, Hazes JM, Dolhain RJ, Bakx PA, Colin EM, Lubberts E (2012) TNF blockade requires 1,25(OH)2D3 to control human Th17-mediated synovial inflammation. Ann Rheum Dis 71:606–612CrossRefPubMedGoogle Scholar
  22. 22.
    Bartok B, Firestein GS (2010) Fibroblast-like synoviocytes: key effector cells in rheumatoid arthritis. Immunol Rev 233:233–255CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Bottini N, Firestein GS (2013) Duality of fibroblast-like synoviocytes in RA: passive responders and imprinted aggressors. Nat Rev Rheumatol 9:24–33CrossRefPubMedGoogle Scholar
  24. 24.
    Mor A, Abramson SB, Pillinger MH (2005) The fibroblast-like synovial cell in rheumatoid arthritis: a key player in inflammation and joint destruction. Clin Immunol 115:118–128CrossRefPubMedGoogle Scholar
  25. 25.
    Beinke S, Ley SC (2004) Functions of NF-kappaB1 and NF-kappaB2 in immune cell biology. Biochem J 382:393–409CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Vallabhapurapu S, Karin M (2009) Regulation and function of NF-kappaB transcription factors in the immune system. Annu Rev Immunol 27:693–733CrossRefPubMedGoogle Scholar
  27. 27.
    Makarov SS (2001) NF-kappa B in rheumatoid arthritis: a pivotal regulator of inflammation, hyperplasia, and tissue destruction. Arthritis Res 3:200–206CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Fu J, Shi Q, Song X, Xia X, Su C, Liu Z, Song E, Song Y (2016) Tetrachlorobenzoquinone exhibits neurotoxicity by inducing inflammatory responses through ROS-mediated IKK/IkappaB/NF-kappaB signaling. Environ Toxicol Pharmacol 41:241–250CrossRefPubMedGoogle Scholar
  29. 29.
    Cohen-Lahav M, Shany S, Tobvin D, Chaimovitz C, Douvdevani A (2006) Vitamin D decreases NFkappaB activity by increasing IkappaBalpha levels. Nephrol Dial Transplant 21:889–897CrossRefPubMedGoogle Scholar

Copyright information

© The Japanese Society for Bone and Mineral Research and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Hua-Qiang Sun
    • 1
  • Dong Yan
    • 1
  • Qin-Nan Wang
    • 2
  • Hong-Zheng Meng
    • 1
  • Ye-Yong Zhang
    • 1
  • Lu-Xu Yin
    • 1
  • Xin-Feng Yan
    • 1
    Email author
  • Shu-Feng Li
    • 1
    Email author
  1. 1.Department of Bone and Joint Surgery, Shandong Provincial Qianfoshan HospitalShandong UniversityJinanPeople’s Republic of China
  2. 2.Department of OrthopedicsWeihai Hospital of Traditional Chinese MedicineWeihaiPeople’s Republic of China

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