Pure endotoxin does not pass across the intestinal epithelium in vitro.

Abstract

Numerous reports suggest that endotoxin (LPS) may play a central role in triggering the inflammatory cascade that leads to the systemic inflammatory response syndrome. Although conditions that promote bacterial translocation in vivo may also facilitate direct translocation of LPS, the exact mechanisms by which LPS crosses the intestinal barrier to reach the systemic circulation are unknown. This study was designed to determine whether pure endotoxin could pass across injured rat ileal mucosa in the Ussing chamber. Sprague-Dawley rats were subjected to mild or severe hemorrhagic shock following carotid artery cannulation, and then resuscitated. Control animals underwent carotid artery cannulation only (sham-shock). Bacterial translocation to the mesenteric lymph nodes, liver, or spleen was measured after 24 h. Transmucosal passage of fluorescein isothiocyanate (FITC)-labeled E. coli C-25, or FITC-conjugated LPS was measured in the Ussing chamber. Intestinal membranes were examined by light and confocal laser microscopy. Severe hemorrhagic shock resulted in a 60% mortality rate and a 100% incidence of bacterial translocation in surviving animals. Sham-shock rats had a 100% survival rate and a 33% incidence of bacterial translocation. Transmucosal passage of FITC-E. coli C-25 was similar in both groups; however, passage of FITC-LPS was never detected. Histologic analysis confirmed mucosal injury to the intestinal epithelium of rats subjected to severe hemorrhagic shock, and confocal laser microscopy demonstrated passage of FITC-E. coil C-25, but not of FITC-LPS across the ileal membranes. Disruption of the intestinal epithelium with a potent mucolytic agent did not result in significant increase in transmucosal passage of FITC-LPS. We conclude that pure LPS does not pass across the intestinal mucosa in vitro. Transmucosal passage of LPS in vivo may be due, at least in part, to the release of bacterial cell wall fragments containing LPS from killed bacteria that had previously translocated.