IL-17A and IL-17F stimulate chemokines via MAPK pathways (ERK1/2 and p38 but not JNK) in mouse cultured mesangial cells: synergy with TNF-alpha and IL-1beta

Am J Physiol Renal Physiol. 2010 Mar;298(3):F779-87. doi: 10.1152/ajprenal.00198.2009. Epub 2009 Dec 30.

Abstract

We investigated the role of IL-17 family members IL-17A and IL-17F in the induction of chemokines in mouse cultured mesangial cells (SV40 MES 13 cells). We evaluated the expression of the chemokines monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) by ELISA and real-time RT-PCR (Q-PCR). Activation of MAPK was assessed by immunoblotting. IL-17RA and IL-17RC were inhibited by small interfering RNA (siRNA). We found that IL-17A or IL-17F stimulation of mesangial cells led to both a dose- and time-dependent increase in MCP-1 and MIP-2 release. This effect was dependent on mRNA transcription and protein translation. Both also enhanced TNF-alpha- and IL-1beta-mediated MCP-1 and MIP-2 release in the cells. Additionally, we observed that IL-17A and IL-17F induced MAPK (p38 MAPK, ERK1/2, and JNK) activation and that pharmacological inhibitors of p38 MAPK (SB203580) and ERK1/2 (U0126), but not JNK (SP600125), blocked the IL-17A/IL-17F-mediated MCP-1 and MIP-2 release. Mesangial cells expressed IL-17RA and IL-17RC, and the IL-17A-mediated MCP-1 and MIP-2 release was significantly blocked by soluble IL-17RA. Furthermore, inhibition of either IL-17RA or IL-17RC expression via siRNA led to significant reduction of IL-17A/IL-17F-stimulated chemokine production. We conclude that IL-17A and IL-17F induce the production of chemokines MCP-1 and MIP-2 via MAPK pathways (p38 MAPK and ERK1/2), as well as mRNA transcription and protein translation and have synergistic effects with TNF-alpha and IL-1beta in cultured mesangial cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Line
  • Chemokine CCL2 / metabolism
  • Chemokine CXCL2 / metabolism
  • Chemokines / genetics
  • Chemokines / metabolism*
  • Enzyme Activation
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Interleukin-17 / metabolism*
  • Interleukin-1beta / metabolism*
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • Mesangial Cells / drug effects
  • Mesangial Cells / enzymology*
  • Mesangial Cells / immunology*
  • Mice
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism*
  • Polymerase Chain Reaction
  • Protein Kinase Inhibitors / pharmacology
  • RNA Interference
  • RNA, Messenger / metabolism
  • Receptors, Interleukin / genetics
  • Receptors, Interleukin / metabolism
  • Receptors, Interleukin-17 / genetics
  • Receptors, Interleukin-17 / metabolism
  • Recombinant Proteins / metabolism
  • Time Factors
  • Transcription, Genetic
  • Tumor Necrosis Factor-alpha / metabolism*
  • Up-Regulation
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Ccl2 protein, mouse
  • Chemokine CCL2
  • Chemokine CXCL2
  • Chemokines
  • Cxcl2 protein, mouse
  • Il17a protein, mouse
  • Il17f protein, mouse
  • Il17ra protein, mouse
  • Il17rc protein, mouse
  • Interleukin-17
  • Interleukin-1beta
  • Protein Kinase Inhibitors
  • RNA, Messenger
  • Receptors, Interleukin
  • Receptors, Interleukin-17
  • Recombinant Proteins
  • Tumor Necrosis Factor-alpha
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases