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1. Intestinal manipulation affects mucosal antimicrobial defense in a mouse model of postoperative ileus.

Author

Kathy Stein, Lena Hieggelke, Bianca Schneiker, Mariola Lysson, Burkhard Stoffels, Sabine Nuding, Jan Wehkamp, Judith Kikhney, Annette Moter, Joerg C Kalff, and Sven Wehner

Subjects
Medicine, Science
Abstract

To explore the effects of abdominal surgery and interleukin-1 signaling on antimicrobial defense in a model of postoperative ileus.C57BL/6 and Interleukin-1 receptor type I (IL-1R1) deficient mice underwent intestinal manipulation to induce POI. Expression of mucosal IL-1α, IL-1β and IL-1R1 and several antimicrobial peptides and enzymes were measured by quantitative PCR or ELISA, western blotting or immunohistochemistry. Bacterial overgrowth was determined by fluorescent in-situ hybridization and counting of jejunal luminal bacteria. Translocation of aerobic and anaerobic bacteria into the intestinal wall, mesenteric lymph nodes, liver and spleen was determined by counting bacterial colonies on agar plates 48h after plating of tissue homogenates. Antimicrobial activity against E. coli and B. vulgatus was analyzed in total and cationic fractions of small bowel mucosal tissue homogenates by a flow cytometry-based bacterial depolarization assay.Jejunal bacterial overgrowth was detected 24h after surgery. At the same time point, but not in the early phase 3h after surgery, bacterial translocation into the liver and mesenteric lymph nodes was observed. Increased antimicrobial activity against E. coli was induced within early phase of POI. Basal antimicrobial peptide and enzyme gene expression was higher in the ileal compared to the jejunal mucosa. The expression of lysozyme 1, cryptdin 1, cryptdin 4 and mucin 2 were reduced 24h after surgery in the ileal mucosa and mucin 2 was also reduced in the jejunum. Postoperative IL-1α and IL-1β were increased in the postoperative mucosa. Deficiency of IL-1R1 affected the expression of antimicrobial peptides during homeostasis and POI.Small bowel antimicrobial capacity is disturbed during POI which is accompanied by bacterial overgrowth and translocation. IL-1R1 is partially involved in the gene expression of mucosal antimicrobial peptides. Altered small bowel antimicrobial activity may contribute also to POI development and manifestation in patients undergoing abdominal surgery.

Published
2018
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2. Intestinal manipulation affects mucosal antimicrobial defense in a mouse model of postoperative ileus

Author

Lena Hieggelke, Joerg C. Kalff, Judith Kikhney, Mariola Lysson, Sven Wehner, Jan Wehkamp, Kathy Stein, Sabine Nuding, Annette Moter, Bianca Schneiker, and Burkhard Stoffels

Subjects
0301 basic medicine, Male, Gene Expression, lcsh:Medicine, Mucin 2, Biochemistry, Mice, Postoperative Complications, Cryptdin, Medicine and Health Sciences, Mesenteric lymph nodes, Intestinal Mucosa, Organ Cultures, lcsh:Science, Multidisciplinary, Antimicrobial, Organoids, medicine.anatomical_structure, Jejunum, Anaerobic bacteria, Biological Cultures, Anatomy, Signal Transduction, Research Article, Antimicrobial peptides, Ileum, Anaerobic Bacteria, Research and Analysis Methods, Microbiology, 03 medical and health sciences, Ileus, medicine, Genetics, Animals, Receptors, Interleukin-1 Type I, Bacteria, business.industry, Mucin, lcsh:R, Organisms, Biology and Life Sciences, Proteins, Gastrointestinal Tract, Disease Models, Animal, 030104 developmental biology, lcsh:Q, business, Digestive System
Abstract

Aim To explore the effects of abdominal surgery and interleukin-1 signaling on antimicrobial defense in a model of postoperative ileus. Methods C57BL/6 and Interleukin-1 receptor type I (IL-1R1) deficient mice underwent intestinal manipulation to induce POI. Expression of mucosal IL-1α, IL-1β and IL-1R1 and several antimicrobial peptides and enzymes were measured by quantitative PCR or ELISA, western blotting or immunohistochemistry. Bacterial overgrowth was determined by fluorescent in-situ hybridization and counting of jejunal luminal bacteria. Translocation of aerobic and anaerobic bacteria into the intestinal wall, mesenteric lymph nodes, liver and spleen was determined by counting bacterial colonies on agar plates 48h after plating of tissue homogenates. Antimicrobial activity against E. coli and B. vulgatus was analyzed in total and cationic fractions of small bowel mucosal tissue homogenates by a flow cytometry-based bacterial depolarization assay. Results Jejunal bacterial overgrowth was detected 24h after surgery. At the same time point, but not in the early phase 3h after surgery, bacterial translocation into the liver and mesenteric lymph nodes was observed. Increased antimicrobial activity against E. coli was induced within early phase of POI. Basal antimicrobial peptide and enzyme gene expression was higher in the ileal compared to the jejunal mucosa. The expression of lysozyme 1, cryptdin 1, cryptdin 4 and mucin 2 were reduced 24h after surgery in the ileal mucosa and mucin 2 was also reduced in the jejunum. Postoperative IL-1α and IL-1β were increased in the postoperative mucosa. Deficiency of IL-1R1 affected the expression of antimicrobial peptides during homeostasis and POI. Conclusion Small bowel antimicrobial capacity is disturbed during POI which is accompanied by bacterial overgrowth and translocation. IL-1R1 is partially involved in the gene expression of mucosal antimicrobial peptides. Altered small bowel antimicrobial activity may contribute also to POI development and manifestation in patients undergoing abdominal surgery.

Published
2018

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