Regulation of interleukin-1beta-induced Chemokine Production and Matrix Metalloproteinase 2 Activation by epigallocatechin-3-gallate in Rheumatoid Arthritis Synovial Fibroblasts

Arthritis Rheum. 2006 Aug;54(8):2393-401. doi: 10.1002/art.22023.

Abstract

Objective: To evaluate the efficacy of epigallocatechin-3-gallate (EGCG), a potent antiinflammatory molecule, in regulating interleukin-1beta (IL-1beta)-induced production of the chemokines RANTES (CCL5), monocyte chemoattractant protein 1 (MCP-1/CCL2), epithelial neutrophil-activating peptide 78 (ENA-78/CXCL5), growth-regulated oncogene alpha (GROalpha/CXCL1), and matrix metalloproteinase 2 (MMP-2) activity in rheumatoid arthritis (RA) synovial fibroblasts.

Methods: Fibroblasts obtained from RA synovium were grown, and conditioned medium was obtained. Cell viability was determined by MTT assay. RANTES, MCP-1, ENA-78, and GROalpha produced in culture supernatants were measured by enzyme-linked immunosorbent assay. MMP-2 activity was analyzed by gelatin zymography. Western blotting was used to study the phosphorylation of protein kinase C (PKC) isoforms and nuclear translocation of NF-kappaB.

Results: EGCG was nontoxic to RA synovial fibroblasts. Treatment with EGCG at 10 microM or 20 microM significantly inhibited IL-1beta-induced ENA-78, RANTES, and GROalpha, but not MCP-1 production in a concentration-dependent manner. EGCG at 50 microM caused a complete block of IL-1beta-induced production of RANTES, ENA-78, and GROalpha, and reduced production of MCP-1 by 48% (P < 0.05). Zymography showed that EGCG blocked constitutive, IL-1beta-induced, and chemokine-mediated MMP-2 activity. Evaluation of signaling events revealed that EGCG preferentially blocked the phosphorylation of PKCdelta and inhibited the activation and nuclear translocation of NF-kappaB in IL-1beta-treated RA synovial fibroblasts.

Conclusion: These results suggest that EGCG may be of potential therapeutic value in inhibiting joint destruction in RA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arthritis, Rheumatoid / metabolism*
  • Arthritis, Rheumatoid / pathology
  • Catechin / analogs & derivatives*
  • Catechin / pharmacology
  • Cell Survival / drug effects
  • Cells, Cultured
  • Chemokines / metabolism*
  • Culture Media, Conditioned / chemistry
  • Culture Media, Conditioned / metabolism
  • Dose-Response Relationship, Drug
  • Down-Regulation
  • Drug Combinations
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Humans
  • Interleukin-1 / pharmacology*
  • Matrix Metalloproteinase 3 / biosynthesis*
  • Synovial Membrane / drug effects*
  • Synovial Membrane / metabolism
  • Synovial Membrane / pathology

Substances

  • Chemokines
  • Culture Media, Conditioned
  • Drug Combinations
  • Interleukin-1
  • Catechin
  • epigallocatechin gallate
  • Matrix Metalloproteinase 3