Abstract
Purpose
Lgr5+ intestinal epithelial stem cells (ISCs) crucial for intestinal epithelial regeneration are impaired during necrotizing enterocolitis. This study aims to investigate the influence of different stressors on intestinal epithelial injury and regeneration in vitro.
Methods
Intestinal epithelial cells (IEC-18) were exposed to stressors such as lipopolysaccharide, hydrogen peroxide, and serum. Cell viability was assessed using MTT assay at 18 and 24 h. IL-6 and Lgr5 gene expressions were measured using qPCR.
Results
IEC-18 cell viability decreased 18 h following administration of lipopolysaccharide, hydrogen peroxide, and low serum concentration. However, after 24 h, the decrease in cell viability was observed only in higher, but not in lower concentrations of lipopolysaccharide and hydrogen peroxide. IL-6 expression increased in all groups compared to control. Lgr5 expression was up-regulated in cells exposed to a single stressor, but down-regulated when multiple stressors were administered.
Conclusion
Lipopolysaccharide, hydrogen peroxide, or low serum induced IEC-18 injury. The upregulation of Lgr5 expression after exposure to a single stressor suggests that minor injury to IEC-18 induces Lgr5+ ISCs to stimulate repair. Conversely, when IEC-18 cells were exposed to multiple stressors, Lgr5 expression was reduced. We speculate that this finding is similar to what happens in NEC when multiple stressors cause impairment of intestinal epithelium regeneration.
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Acknowledgements
This work is supported by Canadian Institutes of Health Research (CIHR) Foundation Grant 353857. AP is the Robert M. Filler Professor of Surgery, The Hospital for Sick Children (HSC), University of Toronto.
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C.L., A.M., B.L., and A.P. designed experiments; C.L., A.M., and B.L. performed experiments, analyzed data, and wrote the manuscript; H.M. and S.S. provided advice and comments; and all the authors revised the manuscript.
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Lee, C., Minich, A., Li, B. et al. Influence of stress factors on intestinal epithelial injury and regeneration. Pediatr Surg Int 34, 155–160 (2018). https://doi.org/10.1007/s00383-017-4183-3
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DOI: https://doi.org/10.1007/s00383-017-4183-3