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Sphingomyelin induces cathepsin D-mediated apoptosis in intestinal epithelial cells and increases inflammation in DSS colitis
  1. Anne Fischbeck1,
  2. Katharina Leucht2,
  3. Isabelle Frey-Wagner2,
  4. Susanne Bentz2,
  5. Theresa Pesch2,
  6. Silvia Kellermeier2,
  7. Michaela Krebs2,
  8. Michael Fried2,
  9. Gerhard Rogler2,
  10. Martin Hausmann2,
  11. Hans-Ulrich Humpf1
  1. 1Institute of Food Chemistry, University of Muenster and NRW Graduate School of Chemistry, Muenster, Germany
  2. 2Clinic of Gastroenterology and Hepatology, Department of Internal Medicine, University Hospital of Zurich, Zürich, Switzerland
  1. Correspondence to Martin Hausmann, Clinic of Gastroenterology and Hepatology, Department of Internal Medicine, University Hospital of Zürich, CH-8091 Zürich, Switzerland; martin.hausmann{at}usz.ch

Abstract

Background The sphingolipid sphingomyelin is a constituent in food derived from animals. Digestive breakdown of sphingomyelin results in ceramide, recently suggested to be involved in activation of cathepsin D as a novel mediator of apoptosis. Damage of the epithelial barrier was detected in patients with inflammatory bowel disease (IBD) due to increased rates of intestinal epithelial cell (IEC) apoptosis.

Methods Acute colitis was induced in C57-BL/6 mice with 2.0% dextran sulfate sodium (DSS) over 7 days. Spontaneous colitis was developed in B6-IL10tm1Cgn (interleukin 10-negative (IL-10–/–)) mice. Mice received 4 or 8 mg sphingomyelin/day by oral gavage. IECs were isolated ex vivo. Apoptosis was determined by propidium iodide (PI) and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining. Execution of apoptosis was confirmed by analysis of active cathepsin D, caspase-3 and caspase-9 with western blot and immunohistochemistry (IHC).

Results Following DSS-mediated colitis, fluorescence-activated cell sorting (FACS) analysis indicated increased apoptosis of IECs under dietary sphingomyelin. The mean sub-G1 portion increased from 8.7±2.5% under a normal diet to 14.0±3.1% under dietary sphingomyelin. Cathepsin activity was significantly increased in isolated IECs after gavage of 4 mg of sphingomyelin per day. Western blot and IHC revealed execution of the apoptotic cascade via activated caspase-3 and caspase-9. Dietary sphingomyelin in the IL-10–/– model confirmed aggravation of mucosal inflammation.

Conclusion Apoptosis of IEC induced by dietary sphingomyelin is mediated via ceramide and cathepsin D activation. This shortens the physiological life cycle of IECs and impairs crucial functions of the intestinal mucosa: barrier, defence and nutrient absorption. The findings provide evidence that dietary sphingomyelin may increase intestinal inflammation.

  • Sphingomyelin
  • cathepsin D
  • intestinal epithelial cells
  • ceramide
  • apoptosis
  • colitis
  • epithelial cells
  • lipids

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Footnotes

  • AF, KL, MH and H-UH contributed equally to this work.

  • Funding This study was supported by grants from the Swiss National Science Foundation (SNF 31003A_127247 to MH and SNF 310030 120312 to GR), by the Deutsche Forschungsgemeinschaft (RO 1236/13-1) and the BMBF Kompetenznetz CED. The Zurich Center for Integrative Human Physiology (ZIHP) to MH and GR, the Swiss inflammatory bowel disease cohort study (SIBDC) to GR and the NRW Graduate School of Chemistry.

  • Competing interests None.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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