Protective effect of the GLP-2 analogue on intestinal ischemia-reperfusion injury in mice

JEREMIAS MOREIRA; PABLO STRINGA; MARÍA VIRGINIA GENTILINI; ARRIOLA BENITEZ, P. CONSTANZA; ARACELI CASTRO; MARTIN RUMBO; GONDOLESI, GABRIEL

Congreso; 18th Congress of the Intestinal Rehabilitation and Transplant Association (CIRTA 2023); 2023

Introduction: The intestinal ischemia-reperfusion injury (IRI) is inherent to the transplant procedure and responsible for direct (early mucosal damage) and indirect injury (lung disease). The loss of mucosal barrier function, the alteration in the permeability and structure of intestinal villi, would release pro-inflammatory cytokines, which cause tissue damage in remote organs. Glucagon-like peptide-2 (GLP-2) is a hormone secreted by enteroendocrine L cells of the intestinal epithelium that has a trophic effect on intestinal epithelium. Although therapeutic use of GLP-2 in intestinal rehabilitation is established, few experimental studies have shown a role in reducing intestinal damage upon IRI. Our aim was to evaluate a possible protective effect of GLP-2 in reducing local and remote organ damage by IRI in a mouse model, as preliminary analysis to be then applied in the transplant model.Methods: An intestinal IRI model clamping the superior mesenteric artery for 40 minutes followed by 30 minutes of reperfusion was used. Five experimental groups were performed: 1) Sham Group without IRI model, 2) Sham Group without IRI model but with intraperitoneal GLP-2 pretreatment for 3 days before surgery and an intraoperative dose of 250µg/kg/day (Sham+GLP2), 3) IRI Control Group (Ct), 4) IRI Group with intraperitoneal GLP-2 pretreatment for 3 days before surgery and an intraoperative dose (GLP2-Pret) and 5) IRI Group with only one intraoperative dose of GLP-2 (GLP2-Intra). After the reperfusion period, mice were sacrificed and intestine and lung samples were obtained. H-E was performed on lung samples to assess tissue damage (degree of alveolar infiltration and hemorrhage among others), and in the intestine to quantify IRI through the Park index and morphological analysis using the Villus/Crypt index. Alcian Blue staining was performed to assess the presence of Goblet cells. One-way ANOVA and Kruskal-Wallis tests were used for statistical analysis.Results: The Sham groups (1 and 2) and GLP2-treated groups (4 and 5), showed less histological intestinal and lung lesions compared to the Ct group (Fig. 1). Significant differences were observed in the number of Goblet cells per villus between the GLP2-Intra (5) (M 11.65 ±1.68) group and the Ct (3) (M 7.68 ±1.89) group (p=0.0156). There were differences between the villus/crypt indices of the Ct group (3) and the GLP2-Intra group (5), but only the differences between the Ct group (3) and the Sham groups (1 and 2) were statistically significant (p=0.0035).Conclusions: A marked decrease in IRI was observed both directly in the intestine and remotely in the Lung using GLP-2 analogue as pretreatment or intraoperatively intraperitoneally. The scarce presence of Goblet cells and a marked morphological alteration of the villi seem to accompany more severe pictures of intestinal IRI damage. According to this study, the use of GLP 2 reduces the IRI opening a new line of research and a potential novel clinical use.