A ceramide analog inhibits cPLA 2 activity and consequent PGE 2 formation in LPS-stimulated macrophages

Meir Goldsmith; Ala Daka; Nadia F. Lamour; Roi Mashiach; Yifat Glucksam; Michael M. Meijler; Charles E. Chalfant; Tsaffrir Zor

doi:10.1016/j.imlet.2010.10.014

A ceramide analog inhibits cPLA ₂ activity and consequent PGE ₂ formation in LPS-stimulated macrophages

Meir Goldsmith, Ala Daka, Nadia F. Lamour, Roi Mashiach, Yifat Glucksam, Michael M. Meijler, Charles E. Chalfant, Tsaffrir Zor

Department of Chemistry

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	136-143
Number of pages	8
Journal	Immunology Letters
Volume	135
Issue number	1-2
DOIs	https://doi.org/10.1016/j.imlet.2010.10.014
State	Published - 30 Mar 2011

Keywords

Arachidonic acid
Ceramide
Ceramide-1-phosphate
LPS
Phospholipase A
Prostaglandin E

ASJC Scopus subject areas

Immunology and Allergy
Immunology

Access to Document

10.1016/j.imlet.2010.10.014

Cite this

@article{54364a4d7af24fc9b05d4385ec542b75,

title = "A ceramide analog inhibits cPLA 2 activity and consequent PGE 2 formation in LPS-stimulated macrophages",

abstract = "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.",

keywords = "Arachidonic acid, Ceramide, Ceramide-1-phosphate, LPS, Phospholipase A, Prostaglandin E",

author = "Meir Goldsmith and Ala Daka and Lamour, {Nadia F.} and Roi Mashiach and Yifat Glucksam and Meijler, {Michael M.} and Chalfant, {Charles E.} and Tsaffrir Zor",

note = "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.",

year = "2011",

month = mar,

day = "30",

doi = "10.1016/j.imlet.2010.10.014",

language = "English",

volume = "135",

pages = "136--143",

journal = "Immunology Letters",

issn = "0165-2478",

publisher = "Elsevier B.V.",

number = "1-2",

}

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

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U2 - 10.1016/j.imlet.2010.10.014

DO - 10.1016/j.imlet.2010.10.014

M3 - Article

AN - SCOPUS:78751705275

SN - 0165-2478

VL - 135

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EP - 143

JO - Immunology Letters

JF - Immunology Letters

IS - 1-2

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