Increased expression and function of integrins in enterocytes by endotoxin impairs epithelial restitution

FG Qureshi, C Leaphart, S Cetin, J Li, A Grishin… - Gastroenterology, 2005 - Elsevier
FG Qureshi, C Leaphart, S Cetin, J Li, A Grishin, S Watkins, HR Ford, DJ Hackam
Gastroenterology, 2005Elsevier
Background & Aims: Experimental necrotizing enterocolitis (NEC) is characterized by
circulating endotoxin (lipopolysaccharide [LPS]) and impaired enterocyte migration. We
hypothesized that LPS increases integrin function and cell-matrix adhesion, leading to
impaired enterocyte migration in the pathogenesis of NEC. Methods: NEC-like intestinal
injury was induced in newborn rats by hypoxia/gavage feedings, and restitution was
determined by assessing bromodeoxyuridine-labeled enterocytes along the crypt-villus axis …
Background & Aims
Experimental necrotizing enterocolitis (NEC) is characterized by circulating endotoxin (lipopolysaccharide [LPS]) and impaired enterocyte migration. We hypothesized that LPS increases integrin function and cell-matrix adhesion, leading to impaired enterocyte migration in the pathogenesis of NEC.
Methods
NEC-like intestinal injury was induced in newborn rats by hypoxia/gavage feedings, and restitution was determined by assessing bromodeoxyuridine-labeled enterocytes along the crypt-villus axis. Newborn mice were injected with 5 mg/kg LPS. IEC-6 cells were treated with LPS ± LY294002 or wortmannin, and β1- and α3-integrins were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunofluorescence. β1-integrin function was determined by adherence of fibronectin beads to IEC-6 monolayers. Migration of IEC-6 cells into a scraped wound was measured by time-lapse microscopy.
Results
Newborn intestinal injury was associated with decreased intestinal restitution and increased α3- and β1-integrin expression in the ileal mucosa, which also was observed after LPS injection. In IEC-6 cells, LPS caused an increase in the expression of α3- and β1-integrins, a shift of β1-integrins from the cytoplasm to the plasma membrane and an increase in fibronectin bead adhesion during which β1-integrins accumulated underneath attached beads. These effects could be reversed with LY294002 or wortmannin, suggesting phosphatidylinositol-3-phosphate kinase (PI3K) dependence. The increased integrin-matrix adhesion by LPS led to an inhibition of enterocyte migration, which could be reversed by anti-β1-antibodies.
Conclusions
Enterocyte migration is inhibited by LPS through increased expression and function of α3- and β1-integrins. Modulation of enterocyte migration via integrins may provide novel insights into the pathogenesis of NEC, in which intestinal restitution is impaired.
Elsevier