Abstract
The interleukin-1β-mitogen-activated protein kinase (MAPK) and NF-κB signaling pathways are involved in the pathogenesis of rheumatoid arthritis. Ebosin, a novel exopolysaccharide (EPS), exhibits anti-inflammatory activity in rat collagen-induced arthritis by suppressing the production of tumor necrosis factor-α, interleukin-6 and interleukin-1β. The aim of the present study was to assess the effects of ebosin on NF-κB and MAPK signaling pathways mediated through interleukin-1β in rat fibroblast-like synoviocytes (FLSs). Western blotting showed decreased production of phosphorylated p38, JNK1, JNK2, IKKα, IKKβ and IκB in the cytoplasm and NF-κB in the nucleus upon ebosin treatment. The DNA-binding activity of NF-κB in the cell nucleus was also inhibited by ebosin treatment, as demonstrated using an electrophoresis mobility gel shift assay. Analysis of the results of the immunofluorescence assay also showed a reduced amount of NF-κB in the nucleus of cells affected by ebosin. These results provided evidence for the effects of ebosin on both interleukin-1β-mediated MAPK and NF-κB signaling pathways in rat FLSs. In addition, enzyme-linked immunosorbent assay demonstrated that ebosin reduces the levels of matrix metalloproteinases MMP-1 and MMP-3 and the chemokines, interleukin-8 and RANTES. Thus, the results of the present study provide further evidence for understanding the medicinal activity of ebosin at a molecular level, therefore nominating this EPS as a potential therapeutic candidate for the treatment of rheumatic arthritis.
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fcmi.2015.36/MediaObjects/41423_2016_Article_BFcmi201536_Fig1_HTML.jpg)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fcmi.2015.36/MediaObjects/41423_2016_Article_BFcmi201536_Fig2_HTML.jpg)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fcmi.2015.36/MediaObjects/41423_2016_Article_BFcmi201536_Fig3_HTML.jpg)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fcmi.2015.36/MediaObjects/41423_2016_Article_BFcmi201536_Fig4_HTML.jpg)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fcmi.2015.36/MediaObjects/41423_2016_Article_BFcmi201536_Fig5_HTML.jpg)
Similar content being viewed by others
References
Feldmann M, Brennan FM, Maini RN . Rheumatoid arthritis. Cell 1996; 85: 307–310.
Burger D, Dayer JM, Palmer G, Gabay C . Is IL-1 a good therapeutic target in the treatment of arthritis? Best Pract Res Clin Rheumatol 2006; 20: 879–896.
Piecyk M, Anderson P . Signal transduction in rheumatoid arthritis. Best Pract Res Clin Rheumatol 2001; 15: 789–803.
Kaminska B . MAPK signalling pathways as molecular targets for anti-inflammatory therapy. Biochim Biophys Acta 2005; 1754: 253–262.
Okamoto H, Cujec TP, Yamanaka H, Kamatani N . Molecular aspects of rheumatoid arthritis: role of transcription factors. FEBS J 2008; 275: 4463–4470.
Okamoto H, Yoshio T, Kaneko H, Yamanaka H . Inhibition of NF-κB signaling by Fasudil as a potential therapeutic strategy for rheumatoid arthritis. Arthritis Rheum 2010; 62: 82–92.
Lee YR, Lee JH, Noh EM, Kim EK, Song MY, Jung WS . Guggulsterone blocks IL-1β-mediated inflammatory responses by suppressing NF-kappaB activation in fibroblast-like synoviocytes. Life Sci 2008; 82: 1203–1209.
Turner-Brannen E, Choi KY, Lippert DN, Cortens JP, Hancock RE, El-Gabalawy H . Modulation of interleukin-1β-induced inflammatory responses by a synthetic cationic innate defence regulator peptide, IDR-1002, in synovial fibroblasts. Arthritis Res Ther 2011; 13: R129.
**g C, Jianbo W, Yuan L, Rong J, Baoyi L . A new IL-1 receptor inhibitor 139A: fermentation, isolation, physico-chemical properties and structure. J Antibiot (Tokyo) 2003; 56: 87–90.
Zhang Y, Wang LF, Bai JY, Guan MJ, Jiang R, Li Y . Anti-inflammatory effect of Ebosin on rat collagen-induced arthritis through suppression production of interferon-1β, interferon-6 and tumor necrosis factor-α. Eur J Inflamm 2013; 11: 677–688.
Cuzzocrea S, Mazzon E, Bevilaqua C, Costantino G, Britti D, Mazzulo G . Cloricromene, a coumarine derivative, protects against collagen-induced arthritis in Lewis rats. Br J Pharmacol 2000; 131: 1399–1407.
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: 248–254.
Yang JP, Merin JP, Nakano T, Kato T, Kitade Y, Okamoto T . Inhibition of the DNA-binding activity of NF-κB by gold compounds in vitro. FEBS Lett 1995; 361: 89–96.
Turner-Brannen E, Choi KY, Lippert DN, Cortens JP, Hancock RE, El-Gabalawy H . Modulation of interleukin-1α-induced inflammatory responses by a synthetic cationic innate defence regulator peptide, IDR-1002, in synovial fibroblasts. Arthritis Res Ther 2011; 13: R129.
Kyriakis JM, Avruch J . Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 2001; 81: 807–869.
Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K . Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions. Endocr Rev 2001; 22: 153–183.
van Loo G, Beyaert R . Negative regulation of NF-κB and its involvement in rheumatoid arthritis. Arthritis Res Ther 2011; 13: 221–231.
DiDonato JA, Mercurio F, Karin M . NF-κB and the link between inflammation and cancer. Immunol Rev 2012; 246: 379–400.
Vincenti MP, Coon CI, Brinckerhoff CE . Nuclear factor κB/p50 activates an element in the distal matrix metalloproteinase 1 promoter in interleukin-1β stimulated synovial fibroblasts. Arthritis Rheum 1998; 41: 1987–1994.
Konttinen YT, Ainola M, Valleala H, Ma J, Ida H, Mandelin J . Analysis of 16 different matrix metalloproteinases (MMP-1 to MMP-20) in the synovial membrane: different profiles in trauma and rheumatoid arthritis. Ann Rheum Dis 1999; 58: 691–697.
Ahmed S, Pakozdi A, Koch AE . Regulation of interleukin-1β-induced chemokine production and matrix metalloproteinase 2 activation by epigallocatechin-3-gallate in rheumatoid arthritis synovial fibroblasts. Arthritis Rheum 2006; 54: 2393–2401.
Monaco C, Andreakos E, Kiriakidis S, Feldmann M, Paleolog E . T-cell-mediated signalling in immune, inflammatory and angiogenic processes: the cascade of events leading to inflammatory diseases. Curr Drug Targets Inflamm Allergy 2004; 3: 35–42.
Schwartsburd PM . Chronic inflammation as inductor of procancer microenvironment: pathogenesis of dysregulated feedback control. Cancer Metastasis Rev 2003; 22: 95–102.
Heller RA, Schena M, Chai A, Shalon D, Bedilion T, Gilmore J . Discovery and analysis of inflammatory disease-related genes using cDNA microarrays. Proc Natl Acad Sci U S A 1997; 94: 2150–2155.
Feldmann M, Maini RN . TNF defined as a therapeutic target for rheumatoid arthritis and other autoimmune diseases. Nat Med 2003; 9: 1245–1250.
Laws A, Gu Y, Marshall V . Biosynthesis, characterization and design of bacterial exopolysaccharides from lactic acid bacteria. Biotechnol Adv 2001; 19: 597–625.
Boels IC, Ramos A, Kleerebezem M, de Vos WM . Functional analysis of the Lactoccus lactis GalU and GalE genes and their impact on sugar nucleotide and exopolysaccharide biosynthesis. Appl Environ Microbiol 2001; 67: 3033–3040.
Boels IC, Kleerebezem M, de Vos WM . Engineering of carbon distribution between glycolysis and sugar nucleotide biosynthesis in Lactococcus lactis. Appl Environ Microbiol 2003; 69: 1129–1135.
Lima LF, Habu S, Gern JC, Nascimento BM, Parada JL, Noseda MD . Production and characterization of the exopolysaccharides produced by Agaricus brasiliensis in submerged fermentation. Appl Biochem Biotechnol 2008; 151: 283–294.
Fernandes MB, Habu S, de Lima MA, Thomaz-Soccol V, Soccol CR . Influence of drying methods over in vitro antitumoral effects of exopolysaccharides produced by Agaricus blazei LPB 03 on submerged fermentation. Bioprocess Biosyst Eng 2011; 34: 253–261.
** M, Lu Z, Huang M, Wang Y, Wang Y . Effects of Se-enriched polysaccharides produced by Enterobacter cloacae Z0206 on alloxan-induced diabetic mice. Int J Biol Macromol 2012; 50: 348–352.
Joo JH, Yun JW . Structure and molecular characterization of extracellular polysaccharides produced by Trichoderma erinaceum DG-312. J Mol Biotechnol 2005; 15: 1250–1257.
Nowak B, Ciszek-Lenda M, Sróttek M, Gamian A, Kontny E, Górska-Fraczek S . Lactobacillus rhamnosus exopolysaccharide ameliorates arthritis induced by the systemic injection of collagen and lipopolysaccharide in DBA/1 mice. Arch Immunol Ther Exp (Warsz) 2012; 60: 211–220.
Sweeney SE, Firestein GS . Signal transduction in rheumatoid arthritis. Curr Opin Rheumatol 2004; 16: 231–237.
Han Z, Boyle DL, Chang L, Bennett B, Karin M, Yang L . c-Jun N-terminal kinase is required for metalloproteinase expression and joint destruction in inflammatory arthritis. J Clin Invest 2001; 108: 73–81.
Inoue T, Hammaker D, Boyle DL, Firestein GS . Regulation of JNK by MKK-7 in fibroblast-like synoviocytes. Arthritis Rheum 2006; 54: 2127–2135.
Shin M, Yan C, Boyd D . An inhibitor of c-jun aminoterminal kinase (SP600125) represses c-Jun activation, DNA-binding and PMA-inducible 92-kDa type IV collagenase expression. Biochim Biophys Acta 2002; 1589: 311–316.
Triantaphyllopoulos K, Madden L, Rioja I, Essex D, Buckton J, Malhotra R . In vitro target validation and in vivo efficacy of p38 MAP kinase inhibition in established chronic collagen-induced arthritis model: a pre-clinical study. Clin Exp Heumatol 2010; 28: 176–185.
Kim EK, Choi EJ . Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta 2010; 1802: 396–405.
Beloueche-Babari M, Jackson LE, Al-Saffar NM, Workman P, Leach MO, Ronen SM . Magnetic resonance spectroscopy monitoring of mitogen-activated protein kinase signaling inhibition. Cancer Res 2005; 65: 3356–3563.
May MJ, Ghosh S . Signal transduction through NF-kappa B. Immunol Today 1998; 19: 80–88.
Sweeney SE, Firestein GS . Rheumatoid arthritis: regulation of synovial inflammation. Int J Biochem Cell Biol 2004; 36: 372–378.
Thornton S, Duwel LE, Boivin GP, Ma Y, Hirsch R . Association of the course of collagen-induced arthritis with distinct patterns of cytokine and chemokine messenger RNA expression. Arthritis Rheum 1999; 42: 1109–1118.
Acknowledgements
This research was financially supported through a grant from the Natural Science Foundation of China (NSFC 30530830) and a grant from the National Key Project of New Drug Study of China (2012ZX09301002-001-023-02).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Zhang, Y., Wang, L., Bai, L. et al. Effect of ebosin on modulating interleukin-1β-induced inflammatory responses in rat fibroblast-like synoviocytes. Cell Mol Immunol 13, 584–592 (2016). https://doi.org/10.1038/cmi.2015.36
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/cmi.2015.36
- Springer Nature Limited
This article is cited by
-
Ebosin: a potential therapeutic agent for rheumatoid arthritis and autoinflammatory syndromes
Cellular & Molecular Immunology (2018)