Yoshifumi Noguchi

Sepsis in rats stimulates cellular proliferation in the mucosa of the small intestine

BACKGROUND/AIMS Increased protein synthesis in intestinal mucosa during sepsis may reflect increased cell turnover. The influence of sepsis and endotoxemia on cellular proliferation in the mucosa of the small intestine was studied. METHODS Sepsis was induced in rats by cecal ligation and puncture. Control rats were sham-operated. Other rats were treated with endotoxin (total dose, 2 mg/kg), human recombinant tumor necrosis factor alpha, or human recombinant interleukin 1 alpha at a dose of 100 micrograms/kg each. Villus height and crypt depth in the jejunum were measured as the number of cells along the side of the villus and crypt, respectively. Cellular proliferation was assessed by measuring the rate of [3H]thymidine incorporation into DNA of the jejunal mucosa and performing autoradiographic studies after intravenous administration of [3H]thymidine. RESULTS Sepsis resulted in reduced villus height, increased crypt depth, and increased incorporation of [3H]thymidine into DNA in the jejunal mucosa. Autoradiography after administration of [3H]thymidine showed labeled cells almost exclusively in the crypts; the number of labeled cells per crpt was higher in septic than in control rats. Administration of endotoxin or recombinant interleukin 1 alpha, but not recombinant tumor necrosis factor alpha, stimulated the incorporation of [3H]-thymidine into DNA in the jejunal mucosa. CONCLUSION Sepsis and endotoxemia stimulate cellular proliferation in the mucosa of the small intestine. This response to sepsis and endotoxemia may be partly mediated by interleukin 1.

Increased glutamine consumption in small intestine epithelial cells during sepsis in rats

BACKGROUND Previous reports have emphasized increased intestinal glutamine consumption during sepsis. This may be of clinical significance as glutamine is a (and perhaps the) physiologic fuel for the intestine. A problem remains, however, as glutaminase, the major enzyme for the degradation of glutamine, is decreased rather than increased in sepsis. This implies an alteration in the metabolic fate of glutamine in sepsis, which may be of clinical significance. METHODS We determined the effect of sepsis on glutamine metabolism in mucosa of small intestine. Sepsis was induced in rats by cecal ligation and puncture. Control rats were sham-operated. After 16 hours, glutamine consumption was measured in isolated enterocytes, and glutaminase, glutamine synthetase, and glutamine transaminase activities and DNA synthesis were determined in mucosa. RESULTS Glutamine consumption was increased during sepsis in enterocytes from the tips of the villi and was unchanged in enterocytes from the midportions of the villi and the crypts. As previously shown, mucosal glutaminase activity was reduced. However, glutamine synthetase and glutamine transaminase activities were stimulated in septic rats, suggesting an increase in metabolism of glutamine via alternate pathways. DNA synthesis was increased during sepsis, especially in crypt cells. CONCLUSIONS Increased or unchanged glutamine consumption in enterocytes from septic rats, despite reduced glutaminase activity, appears to reflect increased activity of other enzyme systems and/or increased utilization of the amino acid for DNA and protein synthesis. Since other aspects of glutamine metabolism may also be deranged in sepsis, a continual enteral supply of glutamine to the intestinal lumen during sepsis may be clinically useful.

Sepsis increases production of total secreted proteins, vasoactive intestinal peptide, and peptide YY in isolated rat enterocytes

The effect of sepsis on the synthesis of endogenous and secretory proteins, including vasoactive intestinal peptide (VIP) and peptide YY (PYY), was determined in enterocytes from jejunum of rats. Sepsis was induced by cecal ligation and puncture (CLP). Control rats were sham-operated. Total endogenous and secreted protein synthesis was assessed in incubated jejunal enterocytes by measuring incorporation of 3H-phenylalanine into protein. Release of VIP and PYY into the medium of incubated enterocytes and cellular levels of the gut peptides were measured by radioimmunoassay. Sixteen hours after CLP, synthesis rates of both endogenous and secreted proteins were increased, and this effect of sepsis was most pronounced in cells from the lower parts of the villi and crypts. Enterocytes from septic rats released more VIP and PYY into the incubation medium, and approximately half of the peptides they released were newly synthesized VIP and PYY. Intracellular levels of VIP and PYY were increased as early as 4 hours after induction of sepsis. Our results suggest that sepsis stimulates the synthesis of endogenous and secretory proteins, including certain gut peptides, in small intestine mucosa. This is consistent with previous observations of increased circulating levels of VIP, PYY and other gastrointestinal hormones during sepsis. The biological significance of increased synthesis of gut peptides and other intestinal proteins during sepsis remains to be determined.

Sepsis increases putrescine concentration and protein synthesis in mucosa of small intestine in rats.

Recent studies suggest that sepsis stimulates mucosal polyamine and protein synthesis. It is not known in which cell type polyamine biosynthesis is increased during sepsis and if polyamines regulate mucosal protein synthesis. We examined the effect of sepsis in rats on polyamine biosynthesis in isolated jejunal enterocytes and measured mucosal protein synthesis following inhibition of ornithine decarboxylase (ODC) activity with difluoromethylornithine. ODC and S-adenosylmethionine decarboxylase (SAMDC) activities and putrescine concentrations were increased in isolated jejunal enterocytes 16 h after induction of sepsis by cecal ligation and puncture. Enterocyte spermidine and spermine levels were not influenced by sepsis. Mucosal ODC and SAMDC activities and polyamine levels were increased following treatment of rats with interleukin-1 but not tumor necrosis factor. Treatment of rats with difluoromethylornithine prevented the sepsis-induced increase in mucosal ODC activity, putrescine concentration, and protein synthesis rate. The results suggest that sepsis increases ODC and SAMDC activities and putrescine concentrations in enterocytes of the small intestine. This metabolic response to sepsis may be regulated by interleukin-1 although other mechanisms may also be involved. Increased mucosal protein synthesis during sepsis may at least in part be regulated by increased putrescine levels.

Influence of sepsis and endotoxemia on polyamine metabolism in mucosa of small intestine in rats

We examined the influence of sepsis and endotoxemia in rats on the biosynthesis of polyamines in small-intestinal mucosa. Sepsis was induced by cecal ligation and puncture (CLP); control rats were sham-operated. In other experiments, rats were treated with two subcutaneous injections of endotoxin (1 mg/kg) or corresponding injections of sterile saline. Ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) activities and concentrations of putrescine, spermidine, and spermine were measured in jejunal mucosa at intervals during 16 hours. Sepsis stimulated ODC and SAMDC activities and increased putrescine and spermidine concentrations in jejunal mucosa. Injection of endotoxin resulted in metabolic changes similar to those observed following CLP. The results suggest that sepsis and endotoxemia stimulate polyamine biosynthesis in mucosa of small intestine. The role of polyamines in the regulation of cell proliferation and metabolic changes in the intestinal mucosa during sepsis remains to be determined.

Differential effects of tumor and parenteral nutrition on jejunal mucosal polyamines.

Nutritional repletion of tumor-bearing (TB) organisms by means of total parenteral nutrition (TPN) has been associated with gut atrophy, immunosuppression, and increased infection rate. To assess possible molecular mechanisms of intestinal atrophy during TPN, jejunal mucosal polyamine concentrations and biosynthetic activity were assessed in non-TB (NTB) and TB rats maintained on rat chow or TPN for eight days. As expected, jejunal mucosal protein content was decreased in both groups of rats maintained on TPN. Although mucosal concentration of putrescine was decreased in TB groups and in the NTB group maintained on TPN, levels of spermidine and spermine were decreased only in the NTB-TPN group. Spermidine levels were elevated significantly in both TB groups. The concentration of spermine was also elevated in the TB-TPN group but was not changed in the TB group maintained on chow. Activity of ornithine decarboxylase was increased in the NTB-TPN group but was not altered significantly in either TB group. S-adenosylmethionine decarboxylase activity was decreased significantly in TB rats maintained on chow and was increased back to control level in the TB-TPN group. These results suggest that jejunal mucosal polyamines are decreased in NTB rats maintained on TPN. Additionally, it appears that enzyme activity is induced in NTB-TPN rats, perhaps in response to the reduction in polyamines and gut atrophy. The absence of similar changes in TB rats maintained on TPN suggests that regulatory mechanisms of polyamine biosynthesis, such as product inhibition, may be refractory. In addition, polyamine biosynthesis from other sources, such as tumor tissue, may be affecting the control of intestinal polyamine biosynthesis.