The Effects and Mechanisms of Sucralfate on Intestinal Epithelial Cells in an In Vitro Model of Necrotizing Enterocolitis

Aim: Necrotizing enterocolitis (NEC) is an important disease which particularly affects premature babies. Although its pathogenesis is not fully explained, it is thought that the mucosal barrier of the intestine is disrupted. Sucralfate is a cytoprotective drug which supports the mucosal barrier. This study aimed to investigate the effects and mechanisms of sucralfate on enterocytes in an in vitro NEC model. Materials and Methods: Intestinal epithelial cells were cultured. NEC was created with lipopolysaccharide (LPS). Sucralfate dose and concentration (106 µL/cm2 2:1 diluted) were determined according to the results of the cell viability test. Control group: This was the cell culture without treatment. Sham group: Only sucralfate was applied to the cell culture. NEC group (NG): Only LPS was applied to the cell culture. Treatment group (TG): the cell culture was first treated with LPS and then sucralfate. Prophylaxis group (PG): First sucralfate and then LPS were applied to the cell culture. Terminal deoxynucleotidyl transferase dUTP end-labeling (TUNEL), caspase 3-8-9, RIPK 1-3, MLKL, occludin, claudin, ICAM and MadCAM were investigated immunohistochemically. Differences between the groups were compared via the one-way ANOVA test. Results: TNF-α and IL-8 levels were higher in the NG (p<0.05). TUNEL positive cells were 65.6±8.2% in the NG and 15.4±3.2% in the TG (p<0.05). Caspase-8,9 and RIPK1 were higher in the NG (p<0.05). The RIPK3 level was low in the NG (p<0.05). MLKL was high in the NG, low in the TG and PG (p<0.05). ICAM-1 was not significantly different between groups. MadCAM-1 was higher in the NG than in the TG and PG (p<0.05). Occludin expression was high and claudin expression was low in the TG (p<0.05). Conclusion: In the vitro NEC model, apoptosis and necroptosis and the expression of cell adhesion molecules change. Sucralfate helps regulate apoptotic - necroptotic activity and cell adhesion molecules. The prophylactic administration of sucralfate does not appear to be as effective as therapeutic administration.