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. 2000 Jul;47(1):112–119. doi: 10.1136/gut.47.1.112

Glucagon-like peptide-2 enhances intestinal epithelial barrier function of both transcellular and paracellular pathways in the mouse

M Benjamin 1, D McKay 1, P Yang 1, H Cameron 1, M Perdue 1
PMCID: PMC1727982  PMID: 10861272

Abstract

BACKGROUND AND AIMS—Glucagon-like peptide-2 (GLP-2) is a recently identified potent intestinotrophic factor. We have evaluated the effect of GLP-2 treatment on intestinal epithelial barrier function in mice.
METHODS—CD-1 mice were injected subcutaneously with GLP-2 or a protease resistant analogue, h[Gly2]GLP-2, twice daily for up to 10 days. Saline injected mice served as controls. Jejunal segments were mounted in Ussing chambers. Tissue conductance was measured and unidirectional fluxes were determined for (i) Na+ and the small inert probe Cr-EDTA (both transported via the paracellular pathway) and (ii) the macromolecule horseradish peroxidase (HRP, transported via the transcellular pathway).
RESULTS—Mice treated with GLP-2 or h[Gly2]GLP-2 for 10 days demonstrated significantly reduced intestinal conductance and fluxes of Na+, Cr-EDTA, and HRP. Electron microscopy confirmed that GLP-2 reduced endocytic uptake of HRP into enterocytes. Functional changes (evident by four hours) preceded morphological changes (evident by 48 hours).
CONCLUSIONS—GLP-2 enhances intestinal epithelial barrier function by affecting both paracellular and transcellular pathways and thus may be of therapeutic value in a number of gastrointestinal conditions.


Keywords: intestinal permeability; macromolecular transport; growth factors

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Figure 1  .

Figure 1  

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(A) Effect of GLP-2 or h[Gly2]GLP-2 treatment (5 µg injected on each of 10 days in mice) on wet weight of the small intestine or small intestinal weight as a percentage of body weight. (B) Effect of GLP-2 or h[Gly2]GLP-2 treatment for 10 days on weight of mucosal scrapings and protein content from a fixed length segment of murine intestine. Values are mean (SEM); n=7-8 mice/group. ***p<0.001 compared with control (phosphate buffered saline (PBS) treated mice); †p<0.05 compared with GLP-2 treated mice.

Figure 2  .

Figure 2  

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Representative photomicrographs of murine intestinal enterocytes. (A) and (C) are from phosphate buffered saline (PBS) treated mice (control); (B) and (D) are from mice after 10 days of treatment with h[Gly2]GLP-2 (5 µg). Note the longer microvilli in h[Gly2]GLP-2 treated mice (B) compared with controls (A) (bar=200 µm). The arrowhead in (B) indicates an endosome containing HRP. Note the increased cell length in h[Gly2]GLP-2 treated mice (D) compared with controls (C) (bar=100 µm).

Figure 3  .

Figure 3  

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Effect of GLP-2 or h[Gly2]GLP-2 treatment (5 µg injected on each of 10 days in mice) on (A) transepithelial ion conductance and (B) flux of 51Cr-EDTA across jejunal segments. Values are mean (SEM). (A) n=30-31 tissues from eight mice per group. (B) n=14-16 tissues from four mice per group.*p<0.05 compared with control (phosphate buffered saline (PBS) treated mice); †p<0.05 compared with GLP-2 treated mice.

Figure 4  .

Figure 4  

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Effect of GLP-2 or h[Gly2]GLP-2 treatment (5 µg injected on each of 10 days in mice) on macromolecular permeability measurements following luminal addition of horseradish peroxidase (HRP 10−5 mol/l). (A) Flux of HRP across jejunal segments. Values are mean (SEM); n=12-15 tissues from 4-5 mice per group.*p<0.05 compared with control mice. (B) Area of HRP containing endosomes in enterocytes 60 minutes after HRP addition. Values are mean (SEM); n=12 photomicrographs per mouse, four mice per group (48 enterocytes). ***p<0.001 compared with control (phosphate buffered saline (PBS) treated mice); †p<0.05 compared with GLP-2 treated mice.

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