Rheumatology International

, Volume 32, Issue 3, pp 729–736 | Cite as

Inhibitors of p38 and ERK1/2 MAPkinase and hydrogen sulphide block constitutive and IL-1β-induced IL-6 and IL-8 expression in the human chondrocyte cell line C-28/I2

Original Article


Mitogen-activated protein kinases (MAPKs) play a central role in inflammatory processes, and their blockage represents pharmacological approaches in the treatment of autoimmune diseases like rheumatoid arthritis (RA). Alternatively, H2S has long been used in sulphur bath therapy for patients suffering from different types of rheumatic disorders, but reports about the beneficial effects of this form of therapy are controversial, rare and of poor scientific quality. The human chondrocyte cell line C-28/I2 was treated with two different MAPK inhibitors (SB203580 and U0126) or with various concentrations of the H2S donor Natrium hydrogen sulphide (NaHS). Thereafter, the secretion of IL-6 and IL-8 was quantified by enzyme-linked immunosorbent assays (ELISAs). The impact of NaHS on the regulation of p38 and ERK1/2 MAPK was confirmed by Western blot experiments. Furthermore, IL-6 and IL-8 expression was quantified by real-time polymerase chain reaction (RT-PCR) and ELISAs from cells which were exposed to SB203580, U0126 and NaHS and stimulated by IL-1β. The C-28/I2 cells constitutively expressed large quantities of IL-6 and IL-8. The data provided prove that in these cells, constitutive as well as IL-1β-induced IL-6 and IL-8 expression was partially and transiently blocked by the treatment of cells with both MAPK inhibitors and NaHS. Presented data seem to be important in evaluating the beneficial functions of MAPK inhibitors and H2S in immune–pathophysiological processes.


Mitogen-activated protein kinases H2Interleukin-6 Interleukin-8 


  1. 1.
    Ivashkiv LB (1996) Cytokine expression and cell activation in inflammatory arthritis. Adv Immunol 63:337–376PubMedCrossRefGoogle Scholar
  2. 2.
    Choy EH, Panayi GS (2001) Cytokine pathways and joint inflammation in rheumatoid arthritis. N Engl J Med 334:907–916Google Scholar
  3. 3.
    Wendling D, Racadot E, Wijdenes J (1993) Treatment of severe rheumatoid arthritis by anti-interleukin 6 monoclonal antibody. J Rheumatol 20:259–262PubMedGoogle Scholar
  4. 4.
    Nakahara H, Nishimoto N (2006) Anti-interleukin-6 receptor antibody therapy in rheumatic diseases. Endocr Metab Immune Disord Drug Targets 6:373–381PubMedGoogle Scholar
  5. 5.
    Cuenda A, Rouse J, Doza YN, Meier R, Cohen P, Gallagher TF, Young PR, Lee JC (1995) SB203580 is a specific inhibitor of a MAP kinase homolog which is stimulated by cellular stresses and interleukin-1. FEBS Lett 364:229–233PubMedCrossRefGoogle Scholar
  6. 6.
    Duncia JV et al (1998) The chemistry and biological activity of U0126, its analogs, and cyclization products. Bioorg Med Chem Lett 8:2839–2844PubMedCrossRefGoogle Scholar
  7. 7.
    Verhagen AP, de Vet HC, de Bie RA, Kessels AG, Boers M, Knipschild PG (1997) Taking baths: the efficacy of balneotherapy in patients with arthritis. A systematic review. J Rheumatol 24:1964–1971PubMedGoogle Scholar
  8. 8.
    Verhagen AP, de Vet HC, de Bie RA, Kessels AG, Boers M, Knipschild PG (2000) Balneotherapy for rheumatoid arthritis and osteoarthritis. Cochrane Database Syst Rev CD000518Google Scholar
  9. 9.
    Verhagen AP, Bierma-Zeinstra SM, Cardoso JR, de Bie RA, Boers M, de Vet HC (2003) Balneotherapy for rheumatoid arthritis. Cochrane Database Syst Rev CD000518Google Scholar
  10. 10.
    Stuhlmeier KM, Bröll J, Iliev B (2009) NF-kappaB independent activation of a series of pro-inflammatory genes by hydrogen sulfide. Exp Biol Med (Maywood) 234(11):1327–1338CrossRefGoogle Scholar
  11. 11.
    Whiteman M, Li L, Rose P, Tan CH, Parkinson DB, Moore PK (2010) The effect of hydrogen sulfide donors on LPS-induced formation of inflammatory mediators in macrophages. Antioxid Redox Signal Feb 3Google Scholar
  12. 12.
    Kloesch B, Liszt M, Broell J (2010) H2S transiently blocks IL-6 expression in rheumatoid arthritic fibroblast-like synoviocytes and deactivates p44/42 mitogen activated protein kinase. Cell Biol Int Jan 12Google Scholar
  13. 13.
    Attene-Ramos MS, Wagner ED, Plewa MJ, Gaskins HR (2006) Evidence that Hydrogen sulfide is a genotoxic agent. Mol Cancer Res 4(1):9–14PubMedCrossRefGoogle Scholar
  14. 14.
    Baskar R, Li L, Moore PK (2007) Hydrogen sulfide induces DNA damage and changes in apoptotic gene expression in human lung fibroblast cells. FASEB J 21:247–255PubMedCrossRefGoogle Scholar
  15. 15.
    Dorman DC, Moulin FJM, McManus BE, Mahle KC, James RA, Struve MF (2002) Cytochrome oxidase inhibition induced by acute hydrogen sulfide inhalation: correlation with tissue sulfide concentrations in the rat brain, liver, lung, and nasal epithelium. Toxicol Sci 65:18–25PubMedCrossRefGoogle Scholar
  16. 16.
    Truong DH, Eghbal MA, Hindmarsh W, Roth SH, O’Brien PJ (2006) Molecular mechanisms of hydrogen sulfide toxicity. Drug Metab Rev 38:733–744PubMedCrossRefGoogle Scholar
  17. 17.
    Zanardo RC, Brancaleone V, Distrutti E, Fiorucci S, Cirino G, Wallace JL (2006) Hydrogen sulfide is an endogenous modulator of leukocyte-mediated inflammation. FASEB J 20:2118–2120PubMedCrossRefGoogle Scholar
  18. 18.
    Szabó C, Kiss L, Pankotai E (2007) Cytoprotective and anti-inflammatory effects of hydrogen sulfide in macrophages and in mice. Crit Care 11(Suppl 2):P2Google Scholar
  19. 19.
    Kimura Y, Dargusch R, Schubert D, Kimura H (2006) Hydrogen sulfide protects HT22 neuronal cells from oxidative stress. Antioxid Redox Signal 8:661–670PubMedCrossRefGoogle Scholar
  20. 20.
    Whiteman M et al (2004) The novel neuromodulator hydrogen sulfide: an endogenous peroxynitrite ‘scavenger’? J Neurochem 90:765–768PubMedCrossRefGoogle Scholar
  21. 21.
    Whiteman M et al (2005) Hydrogen sulfide: a novel inhibitor of hypochlorous acid-mediated oxidative damage in the brain? Biochem Biophys Res Commun 326:794–798PubMedCrossRefGoogle Scholar
  22. 22.
    Yan SK et al (2006) Effects of hydrogen sulfide on homocysteine-induced oxidative stress in vascular smooth muscle cells. Biochem Biophys Res Commun 361:485–491CrossRefGoogle Scholar
  23. 23.
    Yang G, Wu L, Wang R (2006) Pro-apoptotic effect of endogenous H2S on human aorta smooth muscle cells. FASEB J 20:553–555PubMedGoogle Scholar
  24. 24.
    Yang G, Cao K, Wu L, Wang R (2004) Cystathionine γ-lyase overexpression inhibits cell proliferation via a H2S-dependent modulation of ERK1/2 phosphorylation and p21Cip/WAK-1. J Biol Chem 279:49199–49205PubMedCrossRefGoogle Scholar
  25. 25.
    Zhi L, Ang AD, Zhang H, Moore PK, Bhatia M (2007) Hydrogen sulfide induces the synthesis of proinflammatory cytokines in human monocyte cell line U937 via the ERK-NF-{kappa}B pathway. J Leukoc Biol 81:1322–1332PubMedCrossRefGoogle Scholar
  26. 26.
    Nagai Y, Tsugane M, Oka J, Kimura H (2004) Hydrogen sulfide induces calcium waves in astrocytes. FASEB J 18:557–559PubMedGoogle Scholar
  27. 27.
    Qingyou Z et al (2004) Impact of hydrogen sulfide on carbon monoxide/heme oxygenase pathway in the pathogenesis of hypoxic pulmonary hypertension. Biochem Biophys Res Commun 317:30–37PubMedCrossRefGoogle Scholar
  28. 28.
    Oh GS et al (2006) Hydrogen sulfide inhibits nitric oxide production and nuclear factor-κB via hemeoxygenase-1 expression in RAW2647 macrophages stimulated with lipopolysaccharide. Free Radic Biol Med 41:106–119PubMedCrossRefGoogle Scholar
  29. 29.
    Ryter SW, Alam J, Choi AM (2006) Heme oxygenase-1/carbon monoxide: from basic science to therapeutic applications. Physiol Rev 86:583–650PubMedCrossRefGoogle Scholar
  30. 30.
    Goldring MB (2004) Immortalization of human articular chondrocytes for generation of stable, differentiated cell lines. Methods Mol Med 100:23–36PubMedGoogle Scholar
  31. 31.
    Goldring MB (2004) Culture of immortalized chondrocytes and their use as models of chondrocyte function. Methods Mol Med 100:37–52PubMedGoogle Scholar
  32. 32.
    Zhao W, Liu M, Kirkwood KL (2008) p38α stabilizes Interleukin-6 mRNA via multiple AU-rich elements. J Biol Chem 283(4):1778–1785PubMedCrossRefGoogle Scholar
  33. 33.
    Vermeulen L, De Wilde G, Van Damme P, Vanden Berghe W, Haegeman G (2003) Transcriptional activation of the NF-κBp65 subunit by mitogen-and stress-activated protein kinase-1 (MSK1). EMBO J 22:1313PubMedCrossRefGoogle Scholar
  34. 34.
    Gerlo S, Haegeman G, Vanden Berghe W (2008) Transcriptional regulation of autocrine IL-6 expression in multiple myeloma cells. Cell Signal 20:1489–1496PubMedCrossRefGoogle Scholar
  35. 35.
    Milanini-Mongiat J, Pouyssegur J, Pages G (2002) Identification of two Sp1 phosphorylation sites for p42/44 mitogen-activated protein kinases: their implication in vascular endothelial growth factor gene transcription. J Biol Chem 277:20631–20639PubMedCrossRefGoogle Scholar
  36. 36.
    Kunsch C, Rosen CA (1993) NF-κB subunit-specific regulation of the interleukin-8 promoter. Mol Cell Biol 13:6137–6146PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Ludwig Boltzmann Institute for Rheumatology and BalneologyViennaAustria
  2. 2.Department of RheumatologyGeneral Hospital of ViennaViennaAustria

Personalised recommendations