TY - JOUR
T1 - A ceramide analog inhibits cPLA 2 activity and consequent PGE 2 formation in LPS-stimulated macrophages
AU - Goldsmith, Meir
AU - Daka, Ala
AU - Lamour, Nadia F.
AU - Mashiach, Roi
AU - Glucksam, Yifat
AU - Meijler, Michael M.
AU - Chalfant, Charles E.
AU - Zor, Tsaffrir
N1 - Funding Information:
This work was supported by a grant from the European Commission ( IRG #021862 to T.Z.) and by grants from Veteran's Administration (VA Merit Review I and a Research Career Scientist Award to C.E.C.) and from the National Institutes of Health ( HL072925 and CA117950 to C.E.C.). We are grateful to Mrs. Nava Silberstein for superb technical assistance. We thank Dr. Hugh Rosen and Dr. Nathanael S. Gray for supply of reagents, and specifically Mr. Peter Ding and Dr. Mark Parnell for chemical synthesis of PCERA-1. We are grateful to Orna Ernst and to Dr. Amiram Raz for helpful advises and for critical reading of the manuscript.
PY - 2011/3/30
Y1 - 2011/3/30
N2 - Prostaglandin E 2 (PGE 2) is an important mediator of the inflammatory response. Phospho-ceramide analogue-1 (PCERA-1), a synthetic phospholipid-like molecule, was previously reported to modulate pro- and anti-inflammatory cytokine production. We show here that PCERA-1 inhibited LPS-stimulated PGE 2 production in RAW264.7 macrophages, without affecting COX-2 expression. Furthermore, PCERA-1 efficiently suppressed arachidonic acid (AA) release in response to LPS. The dephosphorylated derivative of PCERA-1, ceramide analogue-1 (CERA-1), mimicked the inhibitory effect of PCERA-1 on AA release and PGE 2 production in macrophages. Inhibition of PGE 2 production by CERA-1 was completely rescued by addition of exogenous AA. Importantly, PCERA-1 and ceramide-1-phosphate (C1P) stimulated the enzymatic activity of cPLA 2α in an in vitro assay, whereas CERA-1 and ceramide inhibited both basal and C1P-stimulated cPLA 2α activity. Collectively, these results indicate that CERA-1 suppresses AA release and subsequent PGE 2 production in LPS-stimulated macrophages by direct interaction with cPLA 2, and suggest that ceramide may similarly counteract C1P effect on cPLA 2 activity in cells. The suppression of PGE 2 production is suggested to contribute to the anti-inflammatory action of PCERA-1.
AB - Prostaglandin E 2 (PGE 2) is an important mediator of the inflammatory response. Phospho-ceramide analogue-1 (PCERA-1), a synthetic phospholipid-like molecule, was previously reported to modulate pro- and anti-inflammatory cytokine production. We show here that PCERA-1 inhibited LPS-stimulated PGE 2 production in RAW264.7 macrophages, without affecting COX-2 expression. Furthermore, PCERA-1 efficiently suppressed arachidonic acid (AA) release in response to LPS. The dephosphorylated derivative of PCERA-1, ceramide analogue-1 (CERA-1), mimicked the inhibitory effect of PCERA-1 on AA release and PGE 2 production in macrophages. Inhibition of PGE 2 production by CERA-1 was completely rescued by addition of exogenous AA. Importantly, PCERA-1 and ceramide-1-phosphate (C1P) stimulated the enzymatic activity of cPLA 2α in an in vitro assay, whereas CERA-1 and ceramide inhibited both basal and C1P-stimulated cPLA 2α activity. Collectively, these results indicate that CERA-1 suppresses AA release and subsequent PGE 2 production in LPS-stimulated macrophages by direct interaction with cPLA 2, and suggest that ceramide may similarly counteract C1P effect on cPLA 2 activity in cells. The suppression of PGE 2 production is suggested to contribute to the anti-inflammatory action of PCERA-1.
KW - Arachidonic acid
KW - Ceramide
KW - Ceramide-1-phosphate
KW - LPS
KW - Phospholipase A
KW - Prostaglandin E
UR - http://www.scopus.com/inward/record.url?scp=78751705275&partnerID=8YFLogxK
U2 - 10.1016/j.imlet.2010.10.014
DO - 10.1016/j.imlet.2010.10.014
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AN - SCOPUS:78751705275
SN - 0165-2478
VL - 135
SP - 136
EP - 143
JO - Immunology Letters
JF - Immunology Letters
IS - 1-2
ER -