Enhancement of ileal adaptation by prednisolone after proximal small bowel resection in the rat.

The effect of prednisolone on the adapted ileum of the rat after jejunal resection was examined. Three weeks after 50% proximal small bowel resection animals were fed pharmacological doses of soluble prednisolone (0.75 mg/kg/day) over a one week period, and killed at four weeks. Animals treated with prednisolone showed significant increases in brush border alpha-glucosidase, leucyl-2-napththylamidase and gamma-glutamyl transferase (P less than 0.01) per unit length of intestine compared with resection alone and transection reanastomosis control groups. This increase was the result of a significant enhancement (P less than 0.01) of brush border digestive enzyme activity per milligram of epithelial cell DNA-that is, per enterocyte-and was associated with a similar increase in enterocyte RNA content. In contrast, the activities of lysosomal and mitochondrial marker enzymes per milligran of DNA were similar in each group. Cell proliferation was not further stimulated by prednisolone. Thus prednisolone can selectively enhance brush border digestive capacity after intestinal resection without increasing cell proliferation. The increase in enterocyte RNA suggests that enzyme induction may be the mechanism of this effect.     

Glucose transport by rat small intestine after extensive small-bowel resection

In rats 50 cm of proximal or distal small intestine were resected, preserving duodenum and terminal ileum. Glucose transport was studied 5–6 weeks later, using everted gut sacs from duodenum, ileum, and also from a midgut segment consisting of intestine located preresection at mid-small intestine. Sham-operated animals served as controls: The inner (serosal) fluid medium in sacs from duodenum and midgut gained glucose; ileal sac serosal medium lost glucose. Proximal resection resulted in significant growth of duodenal and midgut mucosa. Duodenal transport specific activity (transport per gram dry mucosa) decreased from control values, but mucosal growth compensated so sac transport capacity (transport per centimeter sac length) remained unchanged. Midgut transport specific activity remained unchanged, thus sac transport capacity directly mirrored increased mucosal mass. Ileal sac serosal medium now accumulated glucose; there was no mucosal growth. Transport specific activity and sac transport capacity of ileum increased in parallel. After distal resection there was no alteration of either duodenal and midgut mucosal masses or transport specific activities, hence sac transport capacities remained unchanged. Ileal sac serosal medium also accumulated glucose, but now both transport specific activity and mucosal mass increased. The resultant increased sac transport capacity was identical to that of ileum after proximal resection. In all sacs from control and resected animals uphill [¹⁴C]glucose concentration differences developed between medium and mucosa. Activity of the mucosal uptake process, assessed in terms of a ratio of mucosal intracellular fluid radioactivity to mucosal medium radioactivity, usually mirrored altered transport specific activity. This indicates that the increased undercoats tissue mass that accompanied increased mucosal mass did not critically affect transport. The most striking findings were: (1) decreased duodenal transport specific activity after proximal resection with mucosal growth compensating; and (2) identical adaptations of ileal segment transport capacities after either proximal or distal small-bowel resections, although mechanisms differed. The present study provides a base for further examinations of carrier-mediated hexose transport after extensive loss of small intestine.     

Evidence for a growth-stimulating fraction in the rat proximal intestine after small bowel resection

Small bowel resection results in a compensatory hyperplasia in the small intestine, but the molecular events that lead to the increased cell production are not known. In this study, a heat-stable acidic extract of the mucosa of the proximal intestine of Sprague-Dawley rats taken 96 h after a 50% small bowel resection was capable of stimulating DNA synthesis of mouse jejunal explants in organ culture. This stimulatory activity was present in the extracts obtained from resected animals after 48 h, 72 h, and 96 h, but was not detectable by 8 days, when presumably a new steady state was established. A significant enhancement of DNA synthesis was observed 96 h after resection when compared with groups of normal and transected animals that were pair-fed with the resected group. This activity was destroyed by protease treatment. Gel filtration experiments showed that the growth-stimulating activity present in the mucosal extract of the 96-h resected animals was due to the presence of two distinct molecules with approximate molecular weights of 4500 and 1500. The extract did not stimulate DNA synthesis in rat peripheral blood lymphocytes, mouse skin fibroblasts, and the colon adenocarcinoma cell line, HCT-8R. Similar extracts taken 96 h after resection from the distal intestine, colon, pancreas, liver, and muscle did not stimulate DNA synthesis of the mouse jejunal explants. These data suggest that the two molecules in the proximal intestine in response to resection could play a role in promoting the observed hyperplasia.     

Pyrimidine biosynthetic enzymes in rat intestine after small bowel resection.

After small bowel resection in the rat, mucosal hyperplasia and an increase in nucleic acid synthesis and cell proliferation occur in remaining small intestine. Male Sprague-Dawley rate underwent resection of 50 cm of proximal or distal intestine or sham operation. One month and 6 months after surgery, aspartate transcarbamylase, dihydroorotase, and uridine kinase were assayed in whole mucosa, and in some instances, in crypt mucosa ffrom the remaining intestinal segment. In control bowel, enzyme activity was significantly greater proximal compared with distal segments. One month after proximal or distal resection, mucosal enzyme activity per cm of gut was greater in the remnant bowel compared with controls. There was no such difference at 6 months. Specific enzyme activity of whole mucosa did not increase after resection because the assay was influenced by the disproportionately large contribution of villous protein. Specific enzyme activity (including thymidine kinase) of isolated crypt mucosa was significantly increased 1 month after distal resection. In addition, [3H]thymidine uptake into DNA of crypt mucosa from proximal remnants was also significantly increased. These results indicate that after small bowel resection, the enzymes of pryimidine biosynthesis increase in remaining bowel and parallel the accelerated rate of cell proliferation.     

Interaction of cholera toxin and membrane GM1 ganglioside of small intestine.

Ganglioside GM1 was isolated from the small intestinal mucosa of man, pig, and beef and amounted to 0.1, 2.0, and 43 nmol per g fresh weight, respectively. These differences in GM1 content were associated with a quantitatively differing ability of the mucosal cells to bind cholera toxin. Human cells bound about 15,000 toxin molecules when saturated with the toxin, porcine cells 120,000, and bovine cells 2,600,000 molecules. The association constant (KA) of the cholera toxin binding was, for cells of all three species, about 10(9) liters/mol. Exogenously added GM1 ganglioside was incorporated in intestinal mucosal cells as well as in intact rabbit small bowel. The increment in GM1 was associated with a correspondingly increased number of binding sites for cholera toxin, whereas KA was unchanged. GM1 incorporation increased the sensitivity of the rabbit small bowel to the diarrheogenic action of cholera toxin. Vibrio cholerae sialidase hydrolyzed isolated intestinal diand trisialogangliosides to GM1. However, the enzyme did not change the ganglioside pattern of intestinal mucosa, had very little influence on the number of toxin binding sites on intestinal cells, and did not alter the sensitivity of the small bowel to the diarrheogenic action of the toxin. These results demonstrate a relationship in the intestinal mucosa between the GM1 ganglioside concentration, the number of binding sites for cholera toxin, and the sensitivity to the biologic action of the toxin. Thus, the study strongly supports the concept that the GM1 ganglioside is the intestinal binding receptor for cholera toxin.     

Amino acid and peptide absorption after proximal small intestinal resection in the rat.

In experimental animals with massive proximal intestinal resection, in vivo ileal absorption of an amino acid mixture containing L-leucine and glycine as well as two different dipeptides, L-leucyl-glycine and glycyl-L-leucine, were compared. Both amino acid and dipeptide absorption were markedly enhanced in the ileal segments. However, the absorption rates from the two perfused dipeptides differed. L-leucine absorption from L-leucyl-glycine was much greater than from glycyl-L-leucine. Brush border amino-peptidase activities after resection were also increased but dissociation between absorption and hydrolytic activity occurred. This study indicates that certain dipeptides are handled differently by adapting ileal segments. Furthermore, the changes observed probably reflect mucosal cellular hyperplasia occurring in association with intestinal adaptation.     

Hypertrophy of the small intestine after its partial resection in the rat

The authors investigated the ultrastructure of small intestinal epithelium in the rat, 3 weeks after resection of the proximal half of the small intestine. They sought to determine: (1) whether the compensatory hypertrophy of the mucosa of the residual small intestine results in changes in the microvilli, in addition to prolongation of the villi; and, (2) whether it is possible to provide evidence of less complete morphologic differentiation in the more rapidly migrating enterocytes. The epithelium on the surface of villi in animals after resection does not differ ultrastructurally from controls and is formed by completely differentiated cellular elements. It is apparent that the compensatory enlargement of the intestinal absorption surface is a result of the hypertrophy of the mucosal villi, and not of changes in the microvilli.     

Effect of serotonin on water and electrolyte transport in the in vivo rabbit small intestine.

The influence of intravenously administered serotonin on water and electrolyte fluxes in the in vivo rabbit jejunum and ileum was examined. Animals were divided into four groups: (1) those receiving saline intravenously while a glucose-free isotonic saline solution perfused the jejunum and ileum; (2) serotonin given intravenously while glucose-free intestinal perfusate was used as in group 1; (3) intravenous saline given while a 10 mM glucose-isotonic saline solution perfused the jejunum and ileum; and (4) intravenous serotonin given while the intestinal perfusate was as in group 3. Serotonin administration resulted in highly significant net secretion of H2O and sodium in both jejunum and ileum in the groups with a glucose-free perfusate. In jejunum, serotonin evoked net water and sodium secretion, whereas controls absorbed water and sodium. In ileum, serotonin significantly enhanced secretion. The addition of glucose to the perfusate completely abolished the serotonin effect. Unidirectional 22Na flux analysis revealed a marked diminution in both mucosal to serosal and serosal to mucosal fluxes in serotonin-treated animals. The decrease in mucosal to serosal flux was greater than the decrease in serosal to mucosal flux, thus explaining the enhanced net secretion observed with serotonin in the groups receiving glucose-free perfusate. In spite of its pronounced effect on water and electrolyte transport, serotonin failed to produce any detectable histological alterations in small bowel mucosa, either by light or electron microscopy. We postulate that serotonin may be an important mediator of the diarrhea so frequently noted in the carcinoid syndrome by virtue of its effects on small intestinal H2O and electrolyte transport.     

Serum proteins in the rat after extensive proximal or distal small bowel resection.

Rats underwent surgical removal of either the proximal or distal half of the small bowel. 5-6 weeks later, total protein concentrations and electrophoretic patterns were determined in sera from the two groups of resected animals. The data compared to similar measurements from sham-operated and unoperated control groups, showed no differences between resected or control animals. This indicates re-establishment of overall nutritional adequacy by this time interval after substantial loss of either proximal or distal small intestine.     

Effect of bicarbonate, acetate, and citrate on water and sodium movement in normal and cholera toxin-treated rat small intestine

Bicarbonate, citrate, or acetate are commonly included in oral rehydration solutions to correct acidosis and possibly because of their ability to promote water and sodium absorption. We have investigated the effect of these anions on water and sodium transport in normal and also in secreting (cholera toxin-treated) rat small intestine using a single-pass perfusion technique. In normal jejunum bicarbonate and acetate produced net absorption, and citrate net secretion of both water and sodium. In normal ileum all anions produced net absorption of water and sodium. In the secreting jejunum, however, bicarbonate had no effect on water and sodium secretion, whereas acetate and citrate actually enhanced the secretory state for both water and sodium. None of these anions had any effect on water and sodium secretion in the ileum. These observations suggest that normal and secreting intestine are qualitatively different with regard to handling of these organic anions. The addition, therefore, of bicarbonate, acetate, or citrate to oral rehydration solutions may have no beneficial effect with regard to the promotion of water and sodium absorption in the secreting intestine during acute diarrhoeal states and could actually be deleterious.     

Effect of lithium ingestion on water and electrolyte transport in rat intestine

The effect of lithium ingestion on intestinal electrolyte and water transport was studied in adult Sprague-Dawley rats. We fed animals a lithium-supplemented diet for 1, 2, 4, or 16 wk before in vivo perfusion of the jejunum and colon. Lithium feeding did not alter jejunal transport of water, electrolytes, or glucose, However, at 4 and 16 wk (16-wk data given) the colon increased net water (168%), sodium (160%), and chloride (140%) absorptions, and the transmural potential difference (396%) as compared with control animals. In addition, the colon absorbed both bicarbonate and potassium against an unfavorable electrochemical gradient. The increased colonic sodium absorption was not associated with an increase in mucosal Na+, K+-ATPase activity. Furthermore, in lithium-fed animals deoxycorticosterone acetate stimulated mucosal Na+, K+-ATPase activity, but it did not further increase net sodium absorption. Neither jejunal nor colonic electrolyte transport was affected 24 h after being gavage-fed lithium. These results suggest that chronic lithium ingestion has a unique mechanism of action as other means of chronically increasing sodium absorption are associated with increased mucosal Na+, K+-ATPase activity.     

Release of vasoactive intestinal polypeptide from the cat small intestine exposed to cholera toxin.

During a four hour observation period vasoactive intestinal polypeptide (VIP) is released in increasing amounts from the feline small intestine exposed to cholera toxin. As VIP is known to be located almost exclusively in the intestinal nerves, the present findings strongly suggest that cholera toxin activates the enteric nervous system. The findings of this and other studies performed in this laboratory lead to the proposal that the choleraic secretion is, at least in part, secondary to the activation of intramural nervous reflexes in the gut.     

Influence of staphylococcal enterotoxin on water and electrolyte transport in the small intestine.

The direct and indirect effects of staphylococcal enterotoxins A and C on the flux of water, sodium, and potassium have been studied in paired Thiry Vella fistulae in dogs. Administration of toxin resulted in a significant decrease in absorption, both in the loop to which the toxin had been administered and its pair. This decrease in absorption was associated with a decrease in movement out of the lumen, movement into the lumen remaining relatively unchanged. The mechanism of action of staphylococcal enterotoxins is discussed, and comparisons made with other enterotoxins.     

Effect of interleukin-2 on microvascular liquid and protein transport in the rat small intestine

Interleukin-2 (IL-2), a glycoprotein lymphokine derived from activated T-lymphocytes displays potent anti-cancer properties but its therapeutic use has been limited by generalized tissue swelling. To shed light on the mechanism underlying this potentially life-threatening edematogenic syndrome, recombinant IL-2 or an equal volume of control solution (excipient or 5% dextrose) was administered to 88 adult, male Sprague-Dawley rats. Initially, rats were injected with 50,000 Cetus units (equal to 300,000 I.U.) of IL-2 intraperitoneally, either one-time ("acute" rats) or every eight hours for two or seven days ("chronic" rats). Thereafter, under pentobarbital anesthesia, the main mesenteric lymph duct was isolated, incised, and measurements made of intestinal lymph flow (JV) and the total protein content and protein fractions in lymph (L) and plasma (P) (refractometry and agarose gel electrophoresis, respectively). Final measurements were also carried out after superior mesenteric vein constriction to assess filtration-independent L/P total protein "washdown." After IL-2, JV and protein clearance (JV x L/P) were increased (p less than 0.001) while lymph and plasma total protein content and protein fractionation were unchanged. Protein washdown was also maintained. These data are not only inconsistent with bulk "plasma leak" from damaged capillaries, but in conjunction with previously demonstrated increased cardiac output and reduced systemic vascular resistance after IL-2 administration, the findings favor augmented microvascular surface exchange area from increased capillary perfusion as the primary mechanism underlying increased transcapillary liquid and protein flux. This conclusion conforms to the rapid reversal of edema in patients after cessation of IL-2 therapy.     

Altered controls of proliferation in proximal small intestine of the senescent rat.

The proximal small intestine responds to starvation by rapidly reducing crypt cell proliferation rate and villus cellularity and to resumption of food intake (refeeding) by abruptly increasing proliferation and the number of villus epithelial cells. We show that villus cellularity responds to starvation and refeeding similarly in young and aging animals. However, as compared to young animals, senescent rats showed increased basal DNA synthetic activity, starvation resulted in a smaller decrease in DNA labeling of crypt cells, and refeeding produced an abrupt broadening of the proliferative zone in older animals without concomitant increased numbers of villus cells. Such altered crypt proliferative responses resemble precancerous changes seen in the colon and the aberrant proliferation found in both small and large intestine after administration of the carcinogen dimethylhydrazine.     

Effect of dopamine and bromocriptine on rat ileal and colonic transport. Stimulation of absorption and reversal of cholera toxin-induced secretion

Water and electrolyte transport were determined in rat ileum and colon using the single-pass perfusion technique. Intraperitoneal dopamine caused prompt stimulation of both ileal and colonic water absorption. The dopamine effect was mediated by both specific dopamine and alpha 2-adrenergic receptors. Haloperidol, a specific dopamine antagonist, and yohimbine, an alpha 2-adrenergic antagonist, inhibited the effect of dopamine in ileal absorption; both antagonists alone had no effect on basal water transport. Bromocriptine (intravenous and intraluminal) stimulated ileal and colonic water absorption, which was inhibited by haloperidol and yohimbine, and reversed cholera toxin-induced ileal secretion. Magnitude and time-courses of the increased water absorption in ileal loops, inoculated with saline, were the same as in loops, inoculated with saline, suggesting that bromocriptine acted to reverse cholera toxin-induced secretion by stimulating absorption. Bromocriptine had no effect on the cyclic adenosine monophosphate increase caused by cholera toxin. We conclude (a) dopamine stimulates water absorption in vivo in rat ileum and colon; (b) this dopamine effect is via specific dopamine and alpha 2-receptors; (c) bromocriptine stimulates water absorption in ileum and colon and also acts by dopamine and alpha 2-receptors; and (d) bromocriptine reverses cholera toxin-induced secretion.     

Increased activity of digestive enzymes in ileal enterocytes adapting to proximal small bowel resection.

The ability of adapting ileal enterocytes to express different digestive enzymes in their brush border membranes was tested in young female Wistar rats (n = 72) receiving 60% proximal small bowel resection. In control rats with intestinal transection both neutral aminopeptidase and alpha-glucosidase activities were shown, by quantitative cytochemistry, to increase during enterocyte migration over the lower part of the villus; thereafter enzyme activities declined or remained approximately constant. Proximal enterectomy increased the amount of alpha-glucosidase but not neutral aminopeptidase activity appearing during early enterocyte development. Thymidine labelled autoradiography showed that the rate of enterocyte migration along the ileal villus nearly doubled after jejunal resection (19.3 v 11.1 microns/h). Nevertheless, the time taken for both peptidase and saccharidase activities to appear at maximal rates in the brush border membrane was diminished by about five hours. Thus ileal enterocytes adapt to proximal small bowel resection by selective increments in enzyme expression, findings that contradict the previous hypothesis of simple metabolic immaturity.     

Bilirubin cycles enterohepatically after ileal resection in the rat

BACKGROUND & AIMS: Patients with ileal disease, resection, or bypass are at increased risk of developing pigment gallstones, but the pathophysiological mechanisms are unknown. The aim of this study was to test the hypothesis that ileectomy induces enterohepatic cycling of bilirubin. METHODS: Ileectomy or sham operation was performed in adult male Sprague-Dawley rats with the following control procedures: no operation, ileal transection, proximal or distal jejunectomy, ileocolonic transposition, and ileocecectomy. Bilirubin and bile salt secretion rates were measured after bile duct cannulation performed 3- 11 days after intestinal surgery. Also measured were bilirubin and bile salt concentrations in the colon as well as indices of hemolysis in blood. RESULTS: Compared with controls, bilirubin secretion rates were increased significantly 3-5 days after ileectomy, distal jejunectomy, ileocolonic transposition, and ileocecectomy, with no hemolysis occurring. Bile salt secretion rates also increased significantly after ileectomy but decreased markedly with prevention of coprophagy, whereas bilirubin secretion rates remained elevated. By 8-11 days after surgery, intestinal adaptation normalized bile salt reabsorption, and hypersecretion of bilirubin was abolished. Colonic levels of unconjugated bilirubin and bile salts were increased fivefold and eightfold respectively in ileectomized animals, but unconjugated bilirubin levels remained normal in bile. CONCLUSIONS: These results are consistent with the hypothesis that enterohepatic cycling of bilirubin occurs with bile salt malabsorption. (Gastroenterology 1996 Jun;110(6):1945-57)     

Role of food in appearance of growth-stimulating activity in rat proximal intestine after small-bowel resection

The oral intake of food is important for the observed compensatory hyperplasia in the remnant small intestine after resection but the molecular events governing this response are not known. Peptides, of molecular weight 4500 and 1000 daltons, present in the proximal intestine for 96 hr after resection and mitogenic for the intestine have been implicated in the adaptive hyperplasia. In this study, the role of food in the appearance of these peptides was assessed. The results show that after resection, rats nourished intravenously demonstrated neither a significant adaptation nor any of the detectable mucosal mitogens, whereas the rats nourished intragastrically demonstrated both hyperplasia and the mitogenic peptide(s). The association of the hyperplasia with the appearance of the mitogenic peptides in the small intestine suggests that they are important in the mechanism by which food promotes the adaptive hyperplasia.This work was supported by a grant from the Medical Research Council of Canada (MA-3320), and from the Justine Lacoste-Beaubien Foundation. This paper was presented in part at the Annual Meeting of the Canadian Society for Clinical Investigation (1986).