Shiga toxin exposure modulates intestinal brush border membrane functional proteins in rabbit ileum

The activities of lactase, sucrase and alkaline phosphatase (AP) were studied in intestinal brush border membranes of control and toxin-treated rabbits. Purified Shiga toxin (Stx) exposure to ileal mucosa inhibited activities of brush border enzymes by 50%. Kinetic analysis revealed that the observed decrease in BBM enzyme activities was due to reduced V(max) with no change in the affinity constants of the systems. The observed changes in enzyme activities were corroborated by Western Blot analysis of lactase, sucrase and AP. The mRNA levels encoding sucrase and lactase proteins in control and Shiga toxin-treated rabbit ileum did not show any change in the rabbit ileum. Histopathological analysis showed short, blunt villi with increased number of inflammatory cells in the lamina propria and extrusion of cells in to the lumen of Stx-treated rabbit ileum. The present findings suggest that Shiga toxin act by inhibiting protein synthesis of these brush border functional proteins beyond their transcriptional level and by the direct damage to intestinal epithelium, which could be implicated in the pathogenesis of diarrhea.