Abstract
In the previous studies, MS80 was found to be able to inhibit the pulmonary fibrosis. However, the target of MS80 remains unclear. To determine the target and the anti-fibrosis mechanisms of MS80, affinity column, MALDI-TOF-MS/MS, co-immunoprecipitation, and co-localization were used. The results showed that MS80 targeting protein was receptor interacting protein 2 (RIP2), which was further confirmed by co-immunoprecipitation and co-localization. Moreover, MS80 inhibited the CD40 ligation-induced NF-κB activation, and subsequently inflammatory cytokines secretion, the collagen synthesis, and the excessive proliferation of fibroblasts. Thus the detailed molecular machinery was ascribed to the involvement of MS80 in targeting CD40 signal pathway via binding and blocking RIP2, the key component of CD40 signal transduction. The findings addressed here may substantially account for the effects of MS80 in combating the pulmonary fibrosis.
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Abbreviations
- SDS-PAGE:
-
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
- BSA:
-
Bovine serum albumin
- FITC:
-
Fluorescein-5-isothiocyanate
- PMSF:
-
Phenyl methyl sulphonyl fluoride
References
Gharaee-Kermani M, Phan SH (2005) Molecular mechanisms of and possible treatment strategies for idiopathic pulmonary fibrosis. Curr Pharm Des 11:3943–3971
Maher TM, Wells AU, Laurent GJ (2007) Idiopathic pulmonary fibrosis: multiple causes and multiple mechanisms? Eur Respir J 30:835–839
Thannickal VJ, Toews GB, White ES et al (2004) Mechanisms of pulmonary fibrosis. Annu Rev Med 55:395–417
Sime PJ, O’Reilly KM (2001) Fibrosis of the lung and other tissues: new concepts in pathogenesis and treatment. Clin Immunol 99:308–319
Kawai M, Masuda A, Kuwana M (2008) A CD40–CD154 interaction in tissue fibrosis. Arthritis Rheum 58:3562–3573
Kaufman J, Graf BA, Leung EC et al (2001) asts as sentinel cells: role of the CDcd40–CDcd40 ligand system in fibroblast activation and lung inflammation and fibrosis. Chest 120:53S–55S
Fukasawa C, Kawaguchi Y, Harigai M et al (2003) Increased CD40 expression in skin fibroblasts from patients with systemic sclerosis (SSc): role of CD40–CD154 in the phenotype of SSc fibroblasts. Eur J Immunol 33:2792–2800
Gelbmann CM, Leeb SN, Vogl D et al (2003) Inducible CD40 expression mediates NFkappaB activation and cytokine secretion in human colonic fibroblasts. Gut 52:1448–1456
Cooper JA Jr (2000) Pulmonary fibrosis: pathways are slowly coming into light. Am J Respir Cell Mol Biol 22:520–523
Schwabe RF, Schnabl B, Kweon YO et al (2001) CD40 activates NF-kappa B and c-Jun N-terminal kinase and enhances chemokine secretion on activated human hepatic stellate cells. J Immunol 166:6812–6819
Kelliher MA, Grimm S, Ishida Y et al (1998) The death domain kinase RIP mediates the TNF-induced NF-kappaB signal. Immunity 8:297–303
McCarthy JV, Ni J, Dixit VM (1998) RIP2 is a novel NF-kappaB-activating and cell death-inducing kinase. J Biol Chem 273:16968–16975
Dixit V, Mak TW (2002) NF-kappaB signaling. Many roads lead to madrid. Cell 111:615–619
Ghosh S, Karin M (2002) Missing pieces in the NF-kappaB puzzle. Cell 109 Suppl:S81–S96
Scheidereit C (2006) IkappaB kinase complexes: gateways to NF-kappaB activation and transcription. Oncogene 25:6685–6705
Komura K, Fujimoto M, Yanaba K et al (2008) Blockade of CD40/CD40 ligand interactions attenuates skin fibrosis and autoimmunity in the tight-skin mouse. Ann Rheum Dis 67:867–872
Kairaitis L, Wang Y, Zheng L et al (2003) Blockade of CD40-CD40 ligand protects against renal injury in chronic proteinuric renal disease. Kidney Int 64:1265–1272
King AE, Kelly RW, Critchley HO et al (2001) Cd40 expression in uterine tissues: a key regulator of cytokine expression by fibroblasts. J Clin Endocrinol Metab 86:405–412
Li L, Bin LH, Li F et al (2005) TRIP6 is a RIP2-associated common signaling component of multiple NF-kappaB activation pathways. J Cell Sci 118:555–563
Ruefli-Brasse AA, Lee WP, Hurst S et al (2004) Rip2 participates in Bcl10 signaling and T-cell receptor-mediated NF-kappaB activation. J Biol Chem 279:1570–1574
Usluoglu N, Pavlovic J, Moelling K et al (2007) RIP2 mediates LPS-induced p38 and IkappaBalpha signaling including IL-12 p40 expression in human monocyte-derived dendritic cells. Eur J Immunol 37:2317–2325
Hasegawa M, Fujimoto Y, Lucas PC et al (2008) A critical role of RICK/RIP2 polyubiquitination in Nod-induced NF-kappaB activation. EMBO J 27:373–383
Chin AI, Dempsey PW, Bruhn K et al (2002) Involvement of receptor-interacting protein 2 in innate and adaptive immune responses. Nature 416:190–194
Kobayashi K, Inohara N, Hernandez LD et al (2002) RICK/Rip2/CARDIAK mediates signalling for receptors of the innate and adaptive immune systems. Nature 416:194–199
Acknowledgments
This work was funded by Natural Science Foundation of China Grants (30271607) and the MALDI-TOF-MS/MS analysis was performed by Research Center for Proteome Analysis, Key Lab of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.
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Du, X., Jiang, S., Liu, H. et al. MS80, a novel sulfated polysaccharide, inhibits CD40-NF-κB pathway via targeting RIP2. Mol Cell Biochem 337, 277–285 (2010). https://doi.org/10.1007/s11010-009-0309-9
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DOI: https://doi.org/10.1007/s11010-009-0309-9