Skip to main content

Effect of P38 mapkinase inhibitor RWJ-67657 on proinflammatory mediators produced by IL-1β-and/or TNFα-stimulated rheumatoid synovial fibroblasts

Introduction

The p38-mitogen activated protein kinase (p38 MAPK) inhibitor RWJ-67657 has been shown to effectively suppress clinical and cytokine response to endotoxin in healthy human volunteers (Fijen JW: JCI 2001, 124:16). In patients with rheumatoid arthritis (RA) p38-MAPK activity is observed in the synovial lining layer (Schett G, Arthritis Rheum 2000, 43:2501). To further characterise the role of p38-MAPK in RA the effect of RWJ-67657 on IL-1β and/or TNFα induced mRNA expression and production of cytokines (IL-6 and IL-8) and matrixmetalloproteinases (MMP-1, MMP-3 and TIMP-1) as well as on mRNA expression of ADAMTS-4 (aggrecanase) and COX-2 was studied in cultured synovial fibroblasts.

Methods

Rheumatoid synovial fibroblasts were isolated from patients who underwent a total joint replacement. Cells at passage 3–8 were stimulated with 1 ng/ml IL-1β and/or TNFα with or without preincubation with 0.001–30 μM RWJ-67657. RNA isolation and RT-PCR was performed after 6 hour stimulation and protein levels in supernatants were measured by ELISA after 48 hour stimulation.

Results

RWJ-67657 induced a dose-related decrease in IL-6, IL-8 and MMP-3 production, both after IL-1β and TNF-α stimulation. Inhibition of MMP-1 was seen only at high levels of RWJ-67657. TIMP-1 was produced constitutively and was not affected by stimulation or inhibition. These findings were all confirmed by mRNA expression studies. COX-2 mRNA expression was induced both by IL-1β and TNFα and could be inhibited by RWJ-67657. ADAMTS-4 mRNA expression was only seen after IL-1β stimulation, which could be inhibited by RWJ-67657.

Conclusion

RWJ-67657 is a potent inhibitor of cytokine and MMP production, as well as of their mRNA expression in stimulated RA synovial fibroblasts. Also COX-2 and aggrecanase mRNA expression was inhibited by RWJ-67657. Thus, inhibition of the p38 MAPK pathway by RWJ-67657 effectively leads to inhibition of different inflammatory mediators produced by rheumatoid synovial fibroblasts. Therefore, RWJ-67657 could be of therapeutic significance in RA.

Author information

Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Westra, J., Harmsen, M., van Rijswijk, M. et al. Effect of P38 mapkinase inhibitor RWJ-67657 on proinflammatory mediators produced by IL-1β-and/or TNFα-stimulated rheumatoid synovial fibroblasts. Arthritis Res Ther 4, 45 (2002). https://doi.org/10.1186/ar487

Download citation

Keywords

  • Rheumatoid Arthritis
  • mRNA Expression
  • Synovial Fibroblast
  • Total Joint Replacement
  • Healthy Human Volunteer