The ceramide-1-phosphate analogue PCERA-1 modulates tumour necrosis factor-alpha and interleukin-10 production in macrophages via the cAMP-PKA-CREB pathway in a GTP-dependent manner.

Abstract

The synthetic phospho-ceramide analogue-1 (PCERA-1) down-regulates production of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha) and up-regulates production of the anti-inflammatory cytokine interleukin-10 (IL-10) in lipopolysaccharide (LPS) -stimulated macrophages. We have previously reported that PCERA-1 increases cyclic adenosine monophosphate (cAMP) levels. The objective of this study was to delineate the signalling pathway leading from PCERA-1 via cAMP to modulation of TNF-alpha and IL-10 production. We show here that PCERA-1 elevates intra-cellular cAMP level in a guanosine triphosphate-dependent manner in RAW264.7 macrophages. The cell-permeable dibutyryl cAMP was able to mimic the effects of PCERA-1 on cytokine production, whereas 8-chloro-phenylthio-methyladenosine-cAMP, which specifically activates the exchange protein directly activated by cAMP (EPAC) but not protein kinase A (PKA), failed to mimic PCERA-1 activities. Consistently, the PKA inhibitor H89 efficiently blocked PCERA-1-driven cytokine modulation as well as PCERA-1-stimulated phosphorylation of cAMP response element binding protein (CREB) on Ser-133. Finally, PCERA-1 activated cAMP-responsive transcription of a luciferase reporter, in synergism with the phosphodiesterase (PDE)-4 inhibitor rolipram. Our results suggest that PCERA-1 activates a G(s) protein-coupled receptor, leading to elevation of cAMP, which acts via the PKA-CREB pathway to promote TNF-alpha suppression and IL-10 induction in LPS-stimulated macrophages. Identification of the PCERA-1 receptor is expected to set up a new target for development of novel anti-inflammatory drugs.

• Authors: Dorit Avni, Amir Philosoph, Michael M Meijler, Tsaffrir Zor
• Journal: Immunology (Immunology) Vol. 129 Issue 3 Pg. 375-85 (Mar 2010) ISSN: 1365-2567 [Electronic] England
• PMID: 19922425 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Anti-Inflammatory Agents, Non-Steroidal
• Ceramides
• Creb1 protein, mouse
• Cyclic AMP Response Element-Binding Protein
• Enzyme Inhibitors
• Imidazoles
• Indoles
• Isoquinolines
• Lipopolysaccharides
• PCERA-1
• Protein Kinase Inhibitors
• Pyridines
• Sulfonamides
• Tumor Necrosis Factor-alpha
• Interleukin-10
• Bucladesine
• Guanosine Triphosphate
• Cyclic AMP
• Ribosomal Protein S6 Kinases, 90-kDa
• mitogen and stress-activated protein kinase 1
• Cyclic AMP-Dependent Protein Kinases
• p38 Mitogen-Activated Protein Kinases
• Adenylyl Cyclases
• Rolipram
• Dinoprostone
• N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide
• SB 203580
• Ro 31-8220

Topics

• Adenylyl Cyclases (metabolism)
• Animals
• Anti-Inflammatory Agents, Non-Steroidal (pharmacology)
• Bucladesine (pharmacology)
• Calcium Signaling (drug effects)
• Cell Line
• Cell Membrane (drug effects, metabolism)
• Ceramides (pharmacology)
• Cyclic AMP (analogs & derivatives, metabolism)
• Cyclic AMP Response Element-Binding Protein (metabolism)
• Cyclic AMP-Dependent Protein Kinases (antagonists & inhibitors, metabolism)
• Dinoprostone (pharmacology)
• Enzyme Inhibitors (pharmacology)
• Genes, Reporter (genetics)
• Guanosine Triphosphate (metabolism)
• Imidazoles (pharmacology)
• Indoles (pharmacology)
• Interleukin-10 (metabolism)
• Isoquinolines (pharmacology)
• Lipopolysaccharides (pharmacology)
• Macrophages (drug effects, metabolism)
• Mice
• Phosphorylation (drug effects)
• Protein Kinase Inhibitors (pharmacology)
• Pyridines (pharmacology)
• Response Elements (genetics)
• Ribosomal Protein S6 Kinases, 90-kDa (antagonists & inhibitors, metabolism)
• Rolipram (pharmacology)
• Signal Transduction (drug effects, physiology)
• Sulfonamides (pharmacology)
• Transfection
• Tumor Necrosis Factor-alpha (metabolism)
• p38 Mitogen-Activated Protein Kinases (antagonists & inhibitors, metabolism)