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

JEREMIAS E. MOREIRA; PABLO STRINGA; MARIA VIRGINIA GENTILINI; CONSTANZA ARRIOLA BENÍTEZ; ARACELI CASTRO; RUMBO MARTIN; GABRIEL E GONDOLESI

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

Introduction: The intestinal ischemia-reperfusion injury (IRI) is inherent to the transplant procedure and responsible for direct (early mucosaldamage) and indirect injury (lung disease). The loss of mucosal barrierfunction, the alteration in the permeability and structure of intestinal villi,would release pro-inflammatory cytokines, which cause tissue damage inremote organs. Glucagon-like peptide-2 (GLP-2) is a hormone secreted byenteroendocrine L cells of the intestinal epithelium that has a trophic effecton intestinal epithelium. Although therapeutic use of GLP-2 in intestinalrehabilitation is established, few experimental studies have shown a role inreducing intestinal damage upon IRI. Our aim was to evaluate a possibleprotective effect of GLP-2 in reducing local and remote organ damageby IRI in a mouse model, as preliminary analysis to be then applied in thetransplant model.Methods: An intestinal IRI model clamping the superior mesentericartery for 40 minutes followed by 30 minutes of reperfusion was used. Fiveexperimental 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 intraoperativedose (GLP2-Pret) and 5) IRI Group with only one intraoperative dose ofGLP-2 (GLP2-Intra). After the reperfusion period, mice were sacrificed andintestine and lung samples were obtained. H-E was performed on lungsamples to assess tissue damage (degree of alveolar infiltration and hemorrhage among others), and in the intestine to quantify IRI through the Parkindex and morphological analysis using the Villus/Crypt index. Alcian Bluestaining was performed to assess the presence of Goblet cells. One-wayANOVA and Kruskal-Wallis tests were used for statistical analysis.Results: The Sham groups (1 and 2) and GLP2-treated groups (4 and5), showed less histological intestinal and lung lesions compared to theCt group (Fig. 1). Significant differences were observed in the number ofGoblet cells per villus between the GLP2-Intra (5) (M 11.65 ±1.68) groupand the Ct (3) (M 7.68 ±1.89) group (p=0.0156). There were differencesbetween the villus/crypt indices of the Ct group (3) and the GLP2-Intragroup (5), but only the differences between the Ct group (3) and the Shamgroups (1 and 2) were statistically significant (p=0.0035).Conclusions: A marked decrease in IRI was observed both directly inthe intestine and remotely in the Lung using GLP-2 analogue as pretreatment or intraoperatively intraperitoneally. The scarce presence of Gobletcells and a marked morphological alteration of the villi seem to accompanymore severe pictures of intestinal IRI damage. According to this study, theuse of GLP 2 reduces the IRI opening a new line of research and a potentialnovel clinical use.