title: The Noncaloric Sweetener Rebaudioside A Stimulates Glucagon-Like Peptide 1 Release and Increases Enteroendocrine Cell Numbers in 2-Dimensional Mouse Organoids Derived from Different Locations of the Intestine.
abstract: BACKGROUND
  Glucagon-like peptide 1 (GLP-1) contributes to satiety and plays a pivotal role in insulin secretion and glucose homeostasis. Similar to GLP-1, peptide YY (PYY) and cholecystokinin also influence food intake. The secretion of these hormones by enteroendocrine cells along the intestine is modulated by nutrients. Preparations from the Stevia rebaudiana plant, including rebaudioside A, are increasingly being used as noncaloric sweeteners.
  
  
  OBJECTIVE
  We investigated the effects of rebaudioside A on enteroendocrine cells by assessing both cell numbers as well as their secretory capacity in an organoid model.
  
  
  METHODS
  A 2-dimensional organoid model derived from duodenal, jejunal, and ileal crypts of a C57BL/6J mouse was developed and characterized with the use of gene expression and immunofluorescence. We stimulated these organoids with 10 mmol/L rebaudioside A for 1 h and measured their GLP-1, PYY, and cholecystokinin release. We also analyzed the effects of rebaudioside A on gene expression in enteroendocrine cells after an 18-h incubation.
  
  
  RESULTS
  The 2-dimensional organoids contained crypt cells and differentiated villus cells, including enterocytes and goblet and enteroendocrine cells. These enteroendocrine cells stained positive for GLP-1, PYY, and serotonin. The cultured 2-dimensional organoids maintained their location-specific gene expression patterns. Compared with the control, rebaudioside A induced GLP-1 secretion 1.7-fold in the duodenum (P < 0.01), 2.2-fold in the jejunum (P < 0.01), and 4.3-fold in the ileum (P < 0.001). PYY release was increased by rebaudioside A 3-fold in the ileum compared with the control (P < 0.05). Long-term (18-h) stimulation with the sweetener induced the expression of the enteroendocrine-specific markers chromogranin A, glucagon, Pyy, and cholecystokinin 3.5- (P < 0.001), 3.5- (P < 0.001), 3.8- (P < 0.05), and 6.5-fold (P < 0.001), respectively.
  
  
  CONCLUSIONS
  These results show novel ex vivo effects of rebaudioside A on enteroendocrine cells of the mouse small intestine and highlight potentially new applications for rebaudioside A in metabolic diseases.
authors: Top Institute Food and Nutrition, Wageningen, Netherlands.|Division of Human Nutrition, Wageningen University, Wageningen, Netherlands; and.
authors: Nikkie
authors: N
authors: van der Wielen
authors: Institute for Life Sciences and Chemistry, Utrecht University of Applied Sciences, Utrecht, Netherlands.
authors: Jean Paul
authors: JP
authors: Ten Klooster
authors: Division of Human Nutrition, Wageningen University, Wageningen, Netherlands; and.
authors: Susanne
authors: S
authors: Muckenschnabl
authors: Institute for Life Sciences and Chemistry, Utrecht University of Applied Sciences, Utrecht, Netherlands.
authors: Raymond
authors: R
authors: Pieters
authors: Top Institute Food and Nutrition, Wageningen, Netherlands.
authors: Henk Fj
authors: HF
authors: Hendriks
authors: Division of Human Nutrition, Wageningen University, Wageningen, Netherlands; and.
authors: Renger F
authors: RF
authors: Witkamp
authors: Division of Human Nutrition, Wageningen University, Wageningen, Netherlands; and jocelijn.meijerink@wur.nl.
authors: Jocelijn
authors: J
authors: Meijerink
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D045786:Diterpenes, Kaurane / Q000494:pharmacology*; D019858:Enteroendocrine Cells / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology; D052216:Glucagon-Like Peptide 1 / Q000235:genetics / Q000378:metabolism*; D007421:Intestine, Small / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D009940:Organoids / Q000187:drug effects* / Q000378:metabolism; D013549:Sweetening Agents / Q000494:pharmacology; D046509:Tissue Culture Techniques
chemical_list: D045786:Diterpenes, Kaurane; D013549:Sweetening Agents; D052216:Glucagon-Like Peptide 1; C025474:rebaudioside A
keywords: GLP-1; duodenum; glucagon-like peptide 1; gut hormone; ileum; jejunum; minigut; organoids; peptide YY; stevia