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The ceramide‐1‐phosphate analogue PCERA‐1 modulates tumour necrosis factor‐α and interleukin‐10 production in macrophages via the cAMP–PKA–CREB pathway in a GTP‐dependent manner
Author(s) -
Avni Dorit,
Philosoph Amir,
Meijler Michael M.,
Zor Tsaffrir
Publication year - 2010
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/j.1365-2567.2009.03188.x
Subject(s) - creb , rolipram , protein kinase a , creb1 , cytokine , cyclic amp response element binding protein , cyclic adenosine monophosphate , microbiology and biotechnology , camp dependent pathway , signal transduction , tumor necrosis factor alpha , biology , phosphodiesterase , kinase , receptor , chemistry , endocrinology , transcription factor , biochemistry , immunology , enzyme , gene
Summary The synthetic phospho‐ceramide analogue‐1 (PCERA‐1) down‐regulates production of the pro‐inflammatory cytokine tumour necrosis factor‐α (TNF‐α) 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‐α 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‐α 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.