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Original article
Ischaemia-induced mucus barrier loss and bacterial penetration are rapidly counteracted by increased goblet cell secretory activity in human and rat colon
  1. Joep Grootjans1,2,
  2. Inca H R Hundscheid1,
  3. Kaatje Lenaerts1,
  4. Bas Boonen1,
  5. Ingrid B Renes3,
  6. Fons K Verheyen4,
  7. Cornelis H Dejong1,
  8. Maarten F von Meyenfeldt1,
  9. Geerard L Beets1,
  10. Wim A Buurman1
  1. 1Department of Surgery, NUTRIM School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, Maastricht, The Netherlands
  2. 2Department of Internal Medicine, Slotervaartziekenhuis, Amsterdam, The Netherlands
  3. 3Laboratory of Paediatrics, Division of Neonatology, Erasmus MC-Sophia, Rotterdam, The Netherlands
  4. 4Department of Molecular Cell Biology and Electron Microscopy, Centre for Research Innovation, Support and Policy (CRISP), Maastricht University Medical Centre, Maastricht, The Netherlands
  1. Correspondence to Dr Joep Grootjans, Department of Surgery, Maastricht University Medical Centre, Universiteitssingel 50, Maastricht 6229 ER, The Netherlands; j.grootjans{at}maastrichtuniversity.nl

Abstract

Objective Colonic ischaemia is frequently observed in clinical practice. This study provides a novel insight into the pathophysiology of colon ischaemia/reperfusion (IR) using a newly developed human and rat experimental model.

Design In 10 patients a small part of colon that had to be removed for surgical reasons was isolated and exposed to 60 min of ischaemia (60I) with/without different periods of reperfusion (30R and 60R). Tissue not exposed to IR served as control. In rats, colon was exposed to 60I, 60I/30R, 60I/120R or 60I/240R (n=7 per group). The tissue was snap-frozen or fixed in glutaraldehyde, formalin or methacarn fixative. Mucins were stained with Periodic Acid Schiff/Alcian Blue (PAS/AB) and MUC2/Dolichos biflorus agglutinin (DBA). Bacteria were studied using electron microscopy (EM) and fluorescent in situ hybridisation (FISH). Neutrophils were studied using myeloperoxidase staining. qPCR was performed for MUC2, interleukin (IL)-6, IL-1β and tumour necrosis factor α.

Results In rats, PAS/AB and MUC2/DBA staining revealed mucus layer detachment at ischaemia which was accompanied by bacterial penetration (in EM and FISH). Human and rat studies showed that, simultaneously, goblet cell secretory activity increased. This was associated with expulsion of bacteria from the crypts and restoration of the mucus layer at 240 min of reperfusion. Inflammation was limited to minor influx of neutrophils and increased expression of proinflammatory cytokines during reperfusion.

Conclusions Colonic ischaemia leads to disruption of the mucus layer facilitating bacterial penetration. This is rapidly counteracted by increased secretory activity of goblet cells, leading to expulsion of bacteria from the crypts as well as restoration of the mucus barrier.

  • Colonic diseases
  • mucosal injury
  • intestinal barrier function
  • ischaemia-reperfusion
  • mucosal barrier
  • immune response
  • apoptosis
  • gastrointestinal surgery
  • inflammatory bowel disorders
  • gut inflammation
  • inflammatory mechanisms
  • intestinal gene regulation
  • nutrition
  • abdominal surgery
  • inflammation
  • infant gut
  • intestinal epithelium
  • epithelial differentiation
  • epithelial barrier
  • epithelial cells
  • small intestine
  • intestinal development
  • epithelial proliferation
  • colorectal cancer

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Footnotes

  • Funding This work was financially supported by the Dutch Digestive Foundation (MLDS grant WO10-57 to CHD).

  • Competing interests None.

  • Ethics approval This study was approved by the Medical Ethical Committee of Maastricht University Medical Centre and was conducted according to the revised version of the Declaration of Helsinki (October 2008, Seoul). Written informed consent was obtained from all patients. Experimental animal protocols were approved by the Animal Ethics Committee of Maastricht University Medical Centre.

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

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