Decreased Capillary Densityin Vivoin Bowel Mucosa of Rats with Normotensive Sepsis

Abstract

Translocation of bacteria and endotoxin leading to sepsis occurs in animals subjected to burns or intestinal ischemia. This may be mediated in part by bowel mucosal microcirculatory dysfunction. However, the direct effect of sepsis on the mucosal microcirculation is unknown. The objective of this study was to develop a technique for intravital microscopy of the mucosa of the small bowel in an animal model of normotensive sepsis. We tested the hypothesis that normotensive sepsis induced by cecal ligation and perforation leads to a decrease in perfused capillaries in the small bowel mucosa at 24 hr. Twelve male Sprague–Dawley rats were hemodynamically monitored and randomly assigned to cecal ligation and perforation (CLP) or control laparotomy (sham). Twenty-four hours after initial surgery each animal was reanesthetized and the mucosal surface of the distal small bowel prepared for intravital microscopy. Laser doppler measurements of bowel wall blood flow were made immediately and repeated after a 30-min stabilization period. Intravital microscopy of the mucosal microcirculation of six villi per animal was performed and the images recorded on videotape (2 min/villus). The areas surrounded by perfused capillaries (intercapillary area) were then measured using video analysis software. Laser doppler flowmetry revealed a decrease in bowel wall blood flow during the stabilization period in the shams that did not occur in the CLP rats. The intercapillary areas were significantly greater in the CLP rats compared to sham rats (1329 ± 316 μm2vs 979 ± 217 μm2,P= 0.044). The intercapillary areas were also more highly variable in the CLP group (median coefficient of variation 102 vs 83% in the sham group,P= 0.025). Intravital microscopy may be used to examine microcirculatory function of the small bowel mucosa. Sepsis induced by CLP leads to a decrease in the number of perfused capillaries in the small bowel mucosa.