Movatterモバイル変換

[0]ホーム
URL:

Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
Thehttps:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log inShow account info
Access keysNCBI HomepageMyNCBI HomepageMain ContentMain Navigation
pubmed logo
Advanced Clipboard
User Guide

Full text links

Nature Publishing Group full text link Nature Publishing Group Free PMC article
Full text links

Actions

Share

.2013 Apr 11;496(7444):238-42.
doi: 10.1038/nature11986. Epub 2013 Mar 24.

Succinate is an inflammatory signal that induces IL-1β through HIF-1α

Affiliations

Succinate is an inflammatory signal that induces IL-1β through HIF-1α

G M Tannahill et al. Nature..

Abstract

Macrophages activated by the Gram-negative bacterial product lipopolysaccharide switch their core metabolism from oxidative phosphorylation to glycolysis. Here we show that inhibition of glycolysis with 2-deoxyglucose suppresses lipopolysaccharide-induced interleukin-1β but not tumour-necrosis factor-α in mouse macrophages. A comprehensive metabolic map of lipopolysaccharide-activated macrophages shows upregulation of glycolytic and downregulation of mitochondrial genes, which correlates directly with the expression profiles of altered metabolites. Lipopolysaccharide strongly increases the levels of the tricarboxylic-acid cycle intermediate succinate. Glutamine-dependent anerplerosis is the principal source of succinate, although the 'GABA (γ-aminobutyric acid) shunt' pathway also has a role. Lipopolysaccharide-induced succinate stabilizes hypoxia-inducible factor-1α, an effect that is inhibited by 2-deoxyglucose, with interleukin-1β as an important target. Lipopolysaccharide also increases succinylation of several proteins. We therefore identify succinate as a metabolite in innate immune signalling, which enhances interleukin-1β production during inflammation.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1. Glycolysis is necessary for LPS-induced IL-1β expression
(A, B) IL-1β and TNFα mRNA in 100ng/ml LPS- or 1×105B pertussis- stimulated BMDMs ± 2DG (1mM) pretreatment for 3h (n=3). (C) Mice i.p. injected ± 2DG (2g/kg) or PBS for 3h, then 15 mg/kg LPS or PBS for 1.5h. Serum levels of IL-1β and TNFα (LPS n=15; LPS+2DG n=14; 2DG n=8; vehicle n=5). Error bars, s.e.m, * p < 0.05; ** p < 0.01.
Fig. 2
Fig. 2. HIF-1α is responsible for LPS-induced IL-1β expression
(A) LPS-induced HIF-1α and IL-1β protein expression ± 2DG (1, 5, 10mM). (B) IL-1β in BMDMs incubated in normoxia or hypoxia for 24h then LPS for 24h. (C) IL-1β mRNA in LPS-stimulated BMDMs pretreated ± αKG derivative (1mM). (D) LPS-induced HIF-1β and IL-1β in BMDMs pretreated ± αKG (0.01, 1, 100, 1000μM). (E) LPS-induced IL-1β protein in WT and HIF-1α-deficient BMDMs. (F) ChIP-PCR using HIF-1α antibody and primers specific for −300 position ofil1b in LPS-treated BMDMs ±2DG (1mM). (G) Reporter activity in RAW-264 cells transfected with il1b- or −357 il1b. Representative of 3 experiments. Error bars, s.d. (I) ChIP-PCR as above in BMDMs treated with LPS ±αKG (1mM). Error bars, s.e.m, ** p < 0.01.
Fig. 3
Fig. 3. Succinate is induced by LPS to drive HIF-1α-induced IL-1β expression
(A) Glucose utilization over time, as a ratio of ECAR:OCR in LPS-treated BMDMs, analysed on the Seahorse XF-24. (B) Succinate abundance in LPS-stimulated BMDMs pretreated ± 2DG (1, 5, 10mM). (C, D) LPS-stimulated BMDMs pretreated ± 5mM diethylsuccinate or butylmalonate (1mM) for 3 h. n=3. (E) Succinyl-lysine expression and corresponding IL-1β in LPS-stimulated BMDMs ± 2DG (1, 5, 10mM) pretreatment for 3h. (J) Average emPAI values (exponentially modified protein abundance index) as a measure of relative abundance of proteins listed in Supplementary Table 1. Error bars, s.e.m, * p< 0.05; ** p< 0.01.
Fig. 4
Fig. 4. Glutamine is the source of LPS-induced succinate
(A) SLC3A2 mRNA in LPS-treated BMDMs for 8 and 24h. (B) IL-1β in human PBMCs transfected with 100nM SLC3A2 siRNA compared non-silencing control. Representative of 3 experiments. (C) GABA abundance in serum-deprived BMDMs stimulated with 10ng/ml LPS for 24h. (D) Schematic representing tracing of13C5,15N2 labelled glutamine in BMDMs treated with LPS for 24h. (E) LPS-stimulated BMDMs pretreated ± vigabatrin (500μM) for 20h then 1mM13C5,15N2 glutamine for 4h. Table represents ratio of13C4-succinate to the sum of all measured succinate isotopomers as measured by LC-MS. The values shown are mean ± s.e.m, n=4. (F, G) LPS-induced HIF-1α and IL-1β protein and IL-1β mRNA (n=7) in BMDMs pretreated ± vigabatrin for 30 min. (H) IL-1β in serum from mice i.p. injected ± vigabatrin (400mg/kg) or PBS for 1.5h, then 15 mg/kg LPS or PBS solution for 1.5h (LPS n=16; LPS+vigabatrin (LPS + V) n=14; vigabatrin (V) n=3; vehicle n=3). (I) IL-1β in serum from mice i.p. injected ± vigabatrin (400mg/kg) or PBS for 1.5h then infected with 1×106Salmonella Typhimurium UK1 i.p. for 2h (J) Survival of mice i.p. injected ± vigabatrin (400mg/kg) or PBS for 1.5h, then 60 mg/kg LPS or PBS (PBS n=10 (not shown), vigabatrin n=10, LPS n=10, LPS + vigabatrin n=10). Error bars, s.e.m, * p < 0.05; ** p < 0.01. (K) Proposed model: LPS induces high levels of succinate to drive HIF-1α-dependent IL-1β expression and protein succinylation.
See this image and copyright information in PMC

Comment in

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Rodriguez-Prados JC, et al. Substrate fate in activated macrophages: a comparison between innate, classic, and alternative activation. J Immunol. 2010;185:605–614. doi: 10.4049/jimmunol.0901698. - DOI - PubMed
    1. Krawczyk CM, et al. Toll-like receptor-induced changes in glycolytic metabolism regulate dendritic cell activation. Blood. 2010;115:4742–4749. doi: 10.1182/blood-2009-10-249540. - DOI - PMC - PubMed
    1. Pan H, Wu X. Hypoxia attenuates inflammatory mediators production induced by Acanthamoeba via Toll-like receptor 4 signaling in human corneal epithelial cells. Biochem Biophys Res Commun. 2012;420:685–691. doi: 10.1016/j.bbrc.2012.03.069. - DOI - PubMed
    1. Hiscott J, et al. Characterization of a functional NF-kappa B site in the human interleukin 1 beta promoter: evidence for a positive autoregulatory loop. Mol Cell Biol. 1993;13:6231–6240. - PMC - PubMed
    1. Ghisletti S, et al. Parallel SUMOylation-dependent pathways mediate gene-and signal-specific transrepression by LXRs and PPARgamma. Mol Cell. 2007;25:57–70. doi: 10.1016/j.molcel.2006.11.022. - DOI - PMC - PubMed

Publication types

MeSH terms

Substances

Related information

Grants and funding

LinkOut - more resources

Full text links
Nature Publishing Group full text link Nature Publishing Group Free PMC article
Cite
Send To

NCBI Literature Resources

MeSHPMCBookshelfDisclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.

[8]ページ先頭
©2009-2025 Movatter.jp