Ceramide 1-phosphate induces macrophage chemoattractant protein-1 release: involvement in ceramide 1-phosphate-stimulated cell migration

Abstract

(C1P) is implicated in inflammatory responses and was recently shown to promote cell migration. However, the mechanisms involved in these actions are poorly described. Using J774A.1 macrophages, we have now discovered a new biological activity of C1P: stimulation of monocyte chemoattractant protein-1 (MCP-1) release. This novel effect of C1P was pertussis toxin (PTX) sensitive, suggesting the intervention of Gi protein-coupled receptors. Treatment of the macrophages with C1P caused activation of the phosphatidylinositol 3-kinase (PI3K)/Akt, mitogen-activated protein kinase kinase (MEK)/extracellularly regulated kinases (ERK), and p38 pathways. Inhibition of these kinases using selective inhibitors or specific siRNA blocked the stimulation of MCP-1 release by C1P. C1P stimulated nuclear factor-B activity, and blockade of this transcription factor also resulted in complete inhibition of MCP-1 release. Also, C1P stimulated MCP-1 release and cell migration in human THP-1 monocytes and 3T3-L1 preadipocytes. A key observation was that sequestration of MCP-1 with a neutralizing antibody or treatment with MCP-1 siRNA abolished C1P-stimulated cell migration. Also, inhibition of the pathways involved in C1P-stimulated MCP-1 release completely blocked the stimulation of cell migration by C1P. It can be concluded that C1P promotes MCP-1 release in different cell types and that this chemokine is a major mediator of C1P-stimulated cell migration. The PI3K/Akt, MEK/ ERK, and p38 pathways are important downstream effectors in this action.ceramide 1-phosphate; monocyte chemoattractant protein-1 release; macrophage migration; sphingosine 1-phosphate REGULATION OF CELL MIGRATION is a complex process involving hundreds of molecules. It is necessary for tissue homeostasis and is crucial for regulation of vital biological processes including embryogenesis, organogenesis, or regeneration (reviewed in Refs. 38, 48). Also, cell migration is fundamental to inflammatory responses (1, 49), but inadequate migratory signals may induce the migration of the wrong cell type to the wrong place, which may have severe effects in the organism. Some examples include autoimmune syndromes, angiogenesis, or the process of metastasis. Although cell migration is a subject of intense investigation, the mechanisms involved in controlling cell movements are incompletely understood. In this connection, growing evidence suggests that some sphingolipids are key metabolites for controlling chemotaxis (52). Our group recently reported that ceramide 1-phosphate (C1P), a sphingolipid metabolite that is present in serum or plasma (25), and which we found to regulate cell growth and survival (15,17,18,20,22), is a chemoattractant molecule for macrophages (23). This finding is particularly relevant when considering that macrophages play critical roles in chronic and acute inflammation and that these cells facilitate cancer cell migration (21, 46). In addition, Barth et al. (4) recently showed that ceramide kinase, the enzyme responsible for the phosphorylation of ce...

Ceramide-1-phosphate inhibits cigarette smoke-induced airway inflammation

“…An explanation for this discrepancy might be the different C1P concentration (1-20 µM) [12,15,33] and the C1P analogue, as well as the different cell types used in these studies. Of note, the solvent in which C1P is dispersed has been reported to be critical for its pro-or anti-inflammatory action.…”

“…However, other studies reported that high concentrations of C16-C1P (15 or 20 µM) stimulated NF-κB activation in the alveolar rat macrophage cell line NR8383 [12] and murine macrophage cell line J774A.1 [15]. Nevertheless, GOLDSMITH et al [14] did not observe any effect of phospho-ceramide analogue-1, a synthetic C1P analogue, on LPS-induced NF-κB activation in the murine macrophage RAW264.7 cell line.…”

“…For instance, C1P stimulates eicosanoid (e.g. prostaglandin E2) biosynthesis, and promotes phagocytosis of neutrophils, migration of macrophages and degranulation of mast cells [15,22,24,28]. Conversely, C1P inhibits LPS-triggered production of pro-inflammatory cytokines such as TNF-α, IL-6 or IL-5/IL-13 in macrophages or mast cells, as well as reducing acid SMase activity [10,22,23,29].…”

“…C16-C1P was acquired from Matreya LLC (Pleasant Gap, PA, USA) and solubilised in sterile nanopure water on ice using a probe sonicator as previously described [15]. Fresh lipopolysaccharide (LPS)-free cigarette smoke extract (CSE) was generated by bubbling smoke from one commercial cigarette (Marlboro; Philip Morris GmbH, Munich, Germany) at a constant rate through 10 mL RPMI with 0.1% fetal calf serum and penicillin/streptomycin.…”

An Endoplasmic Reticulum Stress-Initiated Sphingolipid Metabolite, Ceramide-1-Phosphate, Regulates Epithelial Innate Immunity by Stimulating β-Defensin Production

“…In contrast to Cer itself, which generates antimitogenic and proapoptotic signals, both S1P and another Cer metabolite, ceramide-1-phosphate (C1P), display promitogenic and antiapoptotic activities, while also stimulating cell motility, wound healing, and cytokine production (17)(18)(19)(20)(21)(22)(23)(24)(25)(26). S1P alters cellular function through either a G protein-coupled family of S1P membrane receptors, or S1P-receptor-independent mechanisms (15,27).…”