Bradykinin-induced IL-6 expression through bradykinin B2 receptor, phospholipase C, protein kinase Cdelta and NF-kappaB pathway in human synovial fibroblasts

Mol Immunol. 2008 Aug;45(14):3693-702. doi: 10.1016/j.molimm.2008.06.007. Epub 2008 Jul 14.

Abstract

Bradykinin (BK) is an inflammatory mediator, and shows elevated levels in regions of severe injury and inflammatory diseases. It has been shown to induce interleukin-6 (IL-6) expression in inflammatory responses in rheumatoid arthritis. We investigated the signaling pathway involved in IL-6 production caused by BK in synovial fibroblasts. BK caused concentration- and time-dependent increases in IL-6 production. By using pharmacological inhibitors or genetic inhibition of the BK receptor, siRNA revealed that B2 but not B1 BK receptors are involved in BK-mediated up-regulation of IL-6. BK-mediated IL-6 production was attenuated by phospholipase C inhibitor (U73122), protein kinase Cdelta inhibitor (rottlerin), NF-kappaB inhibitor (PDTC), IkappaB protease inhibitor (TPCK) and NF-kappaB inhibitor peptide. Stimulation of synovial fibroblasts with BK activated IkappaB kinase alpha/beta (IKK alpha/beta), IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 phosphorylation at Ser(276), p65 and p50 translocation from the cytosol to the nucleus and kappaB-luciferase activity. BK mediated an increase of IKK alpha/beta and IkappaBalpha phosphorylation, kappaB-luciferase activity and p65 and p50 binding to the NF-kappaB element was inhibited by B2 BK receptor antagonist (HOE140), U73122 and rottlerin. Our results suggest that BK increased IL-6 production in synovial fibroblasts via the B2 BK receptor/PI-PLC/PKCdelta/and NF-kappaB signaling pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bradykinin / pharmacology*
  • Bradykinin B2 Receptor Antagonists
  • Cells, Cultured
  • Fibroblasts / metabolism*
  • Genes, Reporter
  • Humans
  • I-kappa B Kinase / physiology
  • Interleukin-6 / biosynthesis*
  • Luciferases / metabolism
  • Models, Biological
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / metabolism
  • Phosphorylation
  • Protein Kinase C-delta / antagonists & inhibitors
  • Protein Kinase C-delta / metabolism
  • RNA, Messenger / analysis
  • Receptor, Bradykinin B2 / metabolism
  • Signal Transduction
  • Synovial Membrane / cytology
  • Transcription Factor RelA / metabolism
  • Transfection
  • Type C Phospholipases / antagonists & inhibitors
  • Type C Phospholipases / metabolism
  • Up-Regulation

Substances

  • Bradykinin B2 Receptor Antagonists
  • Interleukin-6
  • NF-kappa B
  • RNA, Messenger
  • Receptor, Bradykinin B2
  • Transcription Factor RelA
  • Luciferases
  • I-kappa B Kinase
  • Protein Kinase C-delta
  • Type C Phospholipases
  • Bradykinin