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J Biol Chem. 2013 Nov 8;288(45):32563-73. doi: 10.1074/jbc.M113.483750. Epub 2013 Sep 30.

Sphingosine kinases are not required for inflammatory responses in macrophages.

Author information

1
From the Center for Vascular Biology, Department of Pathology and Laboratory Medicine.

Abstract

Sphingosine kinases (Sphks), which catalyze the formation of sphingosine 1-phosphate (S1P) from sphingosine, have been implicated as essential intracellular messengers in inflammatory responses. Specifically, intracellular Sphk1-derived S1P was reported to be required for NFκB induction during inflammatory cytokine action. To examine the role of intracellular S1P in the inflammatory response of innate immune cells, we derived murine macrophages that lack both Sphk1 and Sphk2 (MΦ Sphk dKO). Compared with WT counterparts, MΦ Sphk dKO cells showed marked suppression of intracellular S1P levels whereas sphingosine and ceramide levels were strongly up-regulated. Cellular proliferation and apoptosis were similar in MΦ Sphk dKO cells compared with WT counterparts. Treatment of WT and MΦ Sphk dKO with inflammatory mediators TNFα or Escherichia coli LPS resulted in similar NFκB activation and cytokine expression. Furthermore, LPS-induced inflammatory responses, mortality, and thioglycolate-induced macrophage recruitment to the peritoneum were indistinguishable between MΦ Sphk dKO and littermate control mice. Interestingly, autophagic markers were constitutively induced in bone marrow-derived macrophages from Sphk dKO mice. Treatment with exogenous sphingosine further enhanced intracellular sphingolipid levels and autophagosomes. Inhibition of autophagy resulted in caspase-dependent cell death. Together, these data suggest that attenuation of Sphk activity, particularly Sphk2, leads to increased intracellular sphingolipids and autophagy in macrophages.

KEYWORDS:

Autophagy; Ceramide; Inflammation; Lysosomes; Macrophages; Sphingolipid; Sphingosine 1-Phosphate

PMID:
24081141
PMCID:
PMC3820889
DOI:
10.1074/jbc.M113.483750
[Indexed for MEDLINE]
Free PMC Article

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