european crystal network workshop

    Ceramides and dihydroceramides levels are associated with the inflammatory response in a murine model of gout

    Background: The metabolic syndrome is strongly associated with gout and hyperuricemia in man. Patients with gout commonly report acute flares after eating particular foods and it is suspected that metabolic changes, apart from serum urate levels, influence the triggering of the inflammatory response to MSU crystals.
    Objectives: By studying the metabolic changes that impact on the response to MSU crystals in a murine model of gout, we hope to identify the dietary effect on metabolites that are associated with the inflammatory response to MSU crystals.

    Methods: Male mice (C57Bl/6) were fed either a normal diet (ND) or high fat diet HFD (DIO D12492) for >6 weeks. During the last 4 weeks of feeding, half the mice in each group were treated with antibiotics (Baytril® 0.25% + co-amoxicillin) to induce a germ-free state. At day 0, mice were injected i-p with 1mg of MSU crystals. Mice were sacrificed 6h after i-p injection. Serum, plasma and peritoneal exudate samples were collected at sacrifice and serum  plasma obtained 3 weeks before sacrifice. Untargeted and targeted liquid chromatography – mass spectrometry based-metabolomics approach was performed on serum and plasma samples. IL1 and IL6 were measured in serum and peritoneal exudates by ELISA. Animal experimental authorization was obtained for these experiments.

    Results: Mice fed a HFD had a greater pIL6 response. Both diet and inflammation (MSU injection) altered sphingolipid levels. No effect of antibiotics was observed. MSU injection induced marked changes in the sphingolipid profile (38.2% of explained variance), while diet had a lesser effect (16.2% of explained variance). Mice fed an HFD had significantly higher sIL6 pIL6 before and pIL6 levels after MSU injection than mice fed a ND. DhCer C16:0 and Cer C16:0 levels decreased significantly after MSU injection. Pre-MSU levels of DhCer C16:0 and Cer C16:0 showed significantly negative correlations with IL6p levels after i-p injection in HFD animals.

    Conclusion: The sphingolipid profile was significantly changed by diet and inflammation. In mice given a HFD, the levels of particular ceramides were significantly correlated with subsequent inflammatory response as assessed by IL6 secretion, suggesting that they may play a role in modulating the inflammatory response to MSU. These same ceramides and their derivatives have been linked to susceptibility to diabetes and cardiovascular diseases 1,2. Further investigation of these sphingolipids in gout in man is warranted.

    References: 1. Wigger L et al. Cell Rep. 2017 Feb 28;18(9):2269-2279. doi: 10.1016/j.celrep.2017.02.019   2. Summers SA. Cell Metab. 2018 Feb 6;27(2):276-280. doi: 10.1016/j.cmet.2017.12.003