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Kinetic analysis of the intestinal iron absorption process in situ. The potential of vascularly autoperfused intestinal loops.

1 Blood sampling from mesenteric venules during absorption in situ is a useful tool to analyse intestinal absorption kinetics and prehepatic metabolism in different sections of the rat small intestine. By use of a micromanipulator, the method can be applied to the duodenum. This part of the small intestine shows the strongest adaptation of non-haem iron absorption to the demand for iron. 2 Iron absorption kinetics was linear in duodenal and jejunal segments. In iron-deficient animals, intestinal iron absorption capacity was increased in the duodenum, while simultaneously determined galactose absorption showed no change. 3 In situ perfusion and cannulation of mesenteric venules in duodenal segments are described. The use of a micromanipulator permits varying the blood volume collected by changing the vertical angle between the cannula and the mesenteric vessel. 4 Intestinal iron absorption rates remained close to constant when blood flow rates were varied by a factor of about ten. Plasma concentrations of absorbed iron vs mesenteric blood flow rates followed a hyperbolic function, as the plasma concentration of absorbed iron in mesenteric venules increased to the same extent as the blood flow decreased. 5 As the plasma transferrin concentration did not change over the experimental period, the concentration of absorbed iron in the mesenteric plasma exceeded the iron-binding capacity of plasma transferrin at low blood flow rates. This observation shows that enhancement of intestinal iron absorption does not require a corresponding increase in plasma iron-binding capacity in the intestinal tissue. 6 Vascularly perfused gut loops were also used to measure prehepatic metabolism, which may influence organotropism of carcinogenic metabolites. Therefore, this type of preparation is likely to find a variety of toxicological applications.

Animals↗

Disruption of the small-intestine mucosal barrier after intestinal occlusion: a study with light and electron microscopy.

It is known that the gut may serve as a reservoir for various microorganisms, which under specific circumstances may intrude into the systemic circulation, causing systemic infections. The aim of the present study was to estimate the "critical time" of disruption of the small-intestine mucosal barrier in conditions of experimentally induced intestinal occlusion, based on the histopathological alterations observed under light and electron microscopy. Forty rabbits underwent small-intestine obstruction through ligation with a nonabsorbable suture. Blood cultures from portal vein and inferior vena cava, as well as cultures from the peritoneal fluid, a hepatic fragment, and a mesenteric lymph node, were obtained before the ligation (0 h). The same cultures were repeated at 4 and 8 h (group A, 20 rabbits) and at 6 and 12 h after the ligation (group B, 20 rabbits). Small-intestine specimens proximal to the occlusion were taken for examination under the optic and electronic microscope in the same time intervals. Five of 20 rabbits of group A died within 4 h and 6 of 20 rabbits of group B died within 6 h after the operation. All anaerobic cultures were negative. All aerobic cultures that became positive developed Escherichia coli colonies. Intestinal epithelium of dead animals was transformed to cuboid with destruction of goblet cells and alteration in secretion of acid polysaccharides. The mucosal appearance of all rabbits that survived 12 hours after ligation was the same. The disruption of the mucosal barrier begins 4 h after complete intestinal occlusion. At 12 h after complete intestinal occlusion, the disruption is total with different degrees of severity.

Animals↗

Intestinal mucosa diamine oxidase activity reflects intestinal involvement in Crohn's disease.

The activity of diamine oxidase (DAO), an enzyme found in the apical villous cells of the small intestine mucosa, should reflect the status of the intestinal mucosa. Our purpose was to determine whether DAO activity in the intestinal mucosa is diminished in patients with Crohn's disease and correlates with the severity of histological changes. Mucosal DAO activity was determined in 42 tissue specimens from patients with Crohn's enteritis (n = 15), Crohn's colitis (n = 9), and ulcerative colitis (n = 11), and from patients with no intestinal disease (n = 7). DAO activity was estimated by the metabolism of [14C]putrescine. Histologic changes were graded on a scale of 0-4. Normal histology was graded as zero, mild edema, and inflammation in the lamina propria as one, crypt abscess formation and inflammation as two, more severe inflammation plus or minus granulomata as three, and most severe inflammation with active ulceration as four. Tissue DAO activity was significantly less in patients with ileitis (4.8 +/- 3.6) compared with those with Crohn's colitis (15.0 +/- 11.6), ulcerative colitis (16.8 +/- 19.7), and normal intestine (17.6 +/- 14.3 U/mg protein/h, p less than 0.05). Intestinal DAO activity showed a positive correlation with the histologic scores. Recurrence of Crohn's disease developed postoperatively in two patients with low tissue DAO activity (1.4 and 2.9 U/mg protein/h). Intestinal DAO activity is diminished in patients with Crohn's ileitis and correlates with the severity of histologic changes. Tissue DAO activity might prove useful in predicting the risk of recurrence or anastomotic complications after resection for Crohn's disease.

Adult↗

[Intestinal ultrasound in rare small and large intestinal diseases].

Intestinal ultrasonography is a meanwhile established and valid diagnostic method in inflammatory bowel disease, diverticulitis, and appendicitis. Little, however, is known about other more rare intestinal diseases. Serving as a tertiary referral center for a broad spectrum of intestinal diseases we therefore report some aspects of ultrasonography in patients with acute and chronic enteritis and colitis of different origin, e.g., bacterial and viral colitis, ileocecal tuberculosis, AIDS-related enteritis, neutropenic colitis, cystic fibrosis, celiac sprue, vasculitis, benign and malignant tumors of the intestine, amyloidosis, ischemic colitis, and radiogenic enteritis. Ultrasonography may display the transformation of the intestinal wall from normal to pathological states both in inflammatory and neoplastic disease. Besides demonstrating the transmural aspect of inflammation it also shows the mesenteric reaction as well as complications such as fistula, abscesses, stenosis, or ileus. Furthermore, in some diseases intestinal ultrasonography may serve as a diagnostic clue if typical patterns of the bowel wall and impaired peristalsis can be demonstrated. This may lead to an important reduction of invasive and expensive procedures. Ultrasonography is of definite help in the follow-up of inflammatory changes of the bowel wall and primarily diagnostic with respect of other entities (e.g., penicillin-induced segmental hemorrhagic colitis). A sonographic differential diagnosis of diseases of the bowel wall on a purely morphological basis, however, is difficult and rather the exception than the rule. The information gained by ultrasonography regarding intestinal disease, however, is as important and valid as e.g., in case of focal lesions of the liver.

Colitis↗

Differential expression of vasoactive intestinal polypeptide receptor 1 and 2 mRNA in murine intestinal T lymphocyte subtypes.

Neuropeptides may exert a variety of effects on the immune cells at both systemic and mucosal immune sites. The immunoregulatory properties refer to the ability of physiological signals and pathways to influence various immune functions. Vasoactive intestinal polypeptide (VIP), a neuropeptide present in high concentration in gut, was studied for its production and receptor expression in intraepithelial and lamina propria T lymphocytes of mouse intestine. Using reverse transcription-polymerase chain reaction (RT-PCR) analysis, it was demonstrated that VIP receptor 1 (VIPR1) was constantly expressed in intraepithelial and lamina propria T lymphocytes from both small and large intestine. In contrast, VIPR2 was identified only in T cells from small intestine. Further studies on purified subpopulations of T lymphocytes indicated the existence of VIPR2 in CD8(+) T cells, but not CD4(+) and CD4CD8 double negative T cells, although all these three subpopulations displayed VIPR1. In addition, VIPR1 mRNA was detected in splenic T lymphocytes, but no signal was obtained for VIPR2 mRNA, even after stimulation of the cells with anti-CD3(epsilon)-chain mAb, phorbol 12-myristate 13-acetate (PMA) and/or VIP. The presence of VIP receptor(s) on intestinal T lymphocytes was supported by the detection of VIP on the cell surface using dual colour immunoflowcytometry. In-vitro treatment with VIP resulted in a tendency towards an increased size of the VIP immunoreactive T cell population and significantly enhanced the average immunofluorescence intensity of the surface labelling. This indicates that the receptors are partially occupied by locally produced VIP in vivo and that more peptide molecules can be bound on the lymphocytes when needed, released and accumulated in higher concentration at the action sites. We failed to detect the expression of VIP mRNA in T lymphocytes, from either intestine or spleen. These observations support that VIP may be an important immune modulator in gut acting through specific receptors on T lymphocytes. The differential mRNA expression of VIP receptor subtypes in cells with different phenotypes and in different immune compartments may suggest diverse regulatory roles of the neuropeptide in immune responses.

Animals↗

Fetal intestinal graft is the best source for intestinal transplantation.

Adult intestinal allografts have demonstrated high immunogenicity in human transplantation, making the search for new and more favorable grafts an actual problem. Accepting that fetal and newborn immune systems are relatively immature, their intestines could be ideal sources for organ donation. The purpose of this study was to compare the immunogenicity of fetal, newborn, and adult intestine for selection of the least antigenic. Using a bidirectional rat model for immunologic responses, 116 small-bowel transplantations were done: 36 fetal, 40 newborn, and 40 adult grafts. Two histocompatibility barriers and different immunosuppression regimes were used. For fetal and newborn intestines, free grafts into the omentum of adult recipients were done; for adult intestines, accessory grafts in adult recipients of the same age, using vascular anastomoses. The diagnosis of graft rejection and graft-versus-host disease (GVHD) was based on histology of hematoxylin and eosin-stained biopsies from target organs. Recipients of fetal and newborn grafts did not show signs of GVHD, while 12% of the adult group did (P < 0.05). Rejection was less severe in fetal and adult (P > 0.05) than in newborn (P < 0.05) intestinal transplantation. Treatment with 10 mg/kg per day cyclosporine prevented rejection in 70% of fetal and 75% of adult grafts, while all newborn grafts were rejected. Under no immunosuppression, or with low doses of cyclosporine (2 mg/kg per day), all groups showed histologic signs of rejection in almost all cases, the fetal intestine being the least affected. Concerning histocompatibility barriers, grafts were usually less damaged in the weaker transplantation subgroups. Our data indicate that fetal intestine is the least immunogenic of the three grafts studied, suggesting that it will be the most suitable tissue for organ donation.

Age Factors↗

Influence of intestinal inhibitory reflex on mesenteric blood flow through an intestinal segment of the dog.

The variations in both tonus and rhythmical motility of the intestinal musculature are known to affect profoundly intestinal blood flow. In the present study, the influence of marked reflex inhibition of intestinal tone on the blood flow through an intestinal segment was investigated in the dog. Experiments were performed under conditions of both normal circulation and cross-circulation. Inhibitory reflex was elicited by elevating the intraluminal pressure of a loop or by electrical stimulation of the intestinal wall. Fluctuations of blood flow during the intestinal inhibitory reflex were observed with normal circulation. Arterial and venous blood flows decreased in the initial period of intestinal relaxation, and recovered within 30 sec although reflex inhibition still remained. This is an autoregulatory escape phenomenon. In successive periods, arterial and venous blood flows decreased again due to "the venous-arteriolar response" and then recovered to the quiescent level. The decrease in venous blood flow corresponded to an increase of the degree of oxygen saturation in the venous blood. The decreases in arterial and venous blood flows were observed during the inhibitory reflex even when the intestinal circulation was maintained by the cross-circulation. Two different patterns, i.e., the parallel and reversed patterns, were distinguished in correlation with venous blood flow and oxygen saturation. The autoregulatory escape phenomenon could not be elicited in the experiments in the cross-circulation system because variation of the systemic blood pressure during inhibitory reflex was limited to +/-4%.

Animals↗

Identification of intestinal-type Barrett's metaplasia by using the intestine-specific protein villin and esophageal brush cytology.

Villin is an actin-binding cytoskeletal protein required for brush-border formation in the normal small intestinal and renal proximal tubule epithelium. Villin is a marker of cell differentiation in small intestinal and renal cell lineages, and recent studies have shown villin to be highly expressed in 100% of intestinal-type Barrett's metaplasias. This epithelium is the single greatest risk factor for developing esophageal adenocarcinoma and arises when the normal esophageal squamous epithelium is replaced by a small intestine-like columnar epithelium after damage by chronic gastroesophageal reflux. In intestinal-type Barrett's metaplasia, the villin protein exhibits a highly characteristic staining pattern in which strong apical, brush-border staining of columnar epithelial cells is observed. In this study, the ability to identify intestinal metaplastic cells by using this distinct villin staining pattern was examined in endoscopic esophageal brushings from patients with confirmed Barrett's metaplasia. Esophageal brushings from 81% (17 of 21) of patients with Barrett's metaplasia demonstrated individual columnar cells with the characteristic villin staining pattern, whereas all normal esophageal squamous cells, blood cells, and gastric columnar cells were negative for villin expression. Northern blot analysis demonstrated villin mRNA expression in Barrett's metaplasia but not in the normal squamous esophagus or gastric mucosa from the same patients. The combined use of villin immunohistochemical analysis and esophageal brush cytology may provide a simple and effective method of detecting intestinal-type Barrett's metaplasia in patients at higher risk for developing this epithelium, such as those experiencing chronic gastroesophageal reflux symptoms.

Barrett Esophagus↗

Electrical activity of an intestinal epithelial cell line: hyperpolarizing responses to intestinal secretagogues.

Cultured epithelial cells (Intestine 407) derived from fetal human small intestine exhibited spontaneous oscillations of membrane potential between the resting level of about -20 mV and the activated level of about -75 mV. The cells were hyperpolarized to the latter level in response to mechanical or electrical stimuli. The hyperpolarizing responses were also elicited by the application of intestinal secretagogues: acetylcholine, histamine, serotonin and vasoactive intestinal polypeptide (VIP). The spontaneous oscillation of membrane potential became prominent and long-lasting in the presence of acetylcholine, histamine, serotonin or VIP. These secretagogue-induced responses were mediated by individual independent receptors on the cell membrane. Muscarinic receptors were responsible for the acetylcholine response, and H1-receptors for the histamine response. The cells also responded with a slow hyperpolarization to calcium ionophore A23187, which is known to induce intestinal secretion. The spontaneously occurring hyperpolarizing responses and those induced by stimuli were both due to an increase in the K+ conductance of the cell membrane. Since acetylcholine, histamine, serotonin and A23187 are known to promote mobilization of cellular Ca2+ ions in intestinal secretory cells, it is hypothesized that these electrical activities of the cell are closely related to the receptor stimulation which leads to the Ca2+-mediated intestinal secretion.

Acetylcholine↗

Effect of streptozotocin-induced diabetes mellitus on release of vasoactive intestinal polypeptide from rodent small intestine.

Representative longitudinal muscle strips (6 x 10 mm) from proximal and distal small intestine were excised from control and streptozotocin-treated rats after one month of untreated and insulin-treated diabetes. Untreated diabetes significantly reduced tissue concentrations of vasoactive intestinal polypeptide (VIP) at both intestinal loci. Insulin treatment of the diabetic animals restored tissue VIP concentrations to control group levels, although the beneficial effect of insulin treatment was only significant in the duodenum. Spontaneous release of VIP was significantly attenuated by untreated diabetes at both intestinal sites. In the duodenum, insulin treatment of the diabetic animals restored VIP release to levels indistinguishable from control group values. In the ileum, insulin treatment produced levels of VIP release that were not significantly different from those of the control and untreated diabetic groups. Tetrodotoxin (5 x 10(-6) M) significantly--but incompletely--inhibited VIP release from control group animals at both intestinal sites. These observations indicate that diabetes mellitus significantly diminishes VIP tissue concentrations and release from intestinal myenteric nerves. These abnormalities improve with insulin treatment. However, the mechanisms of VIP release from proximal and distal intestine appear to differ not only in their response to the diabetic state, but also in their response to insulin treatment.

Animals↗

The role of intestinal transplantation in the management of intestinal failure.

Significantly reduced morbidity and mortality is needed before intestinal transplantation will be applicable in most patients with intestinal failure who are on long-term total parenteral nutrition (TPN). However, transplantation does play a role if TPN fails, with failure defined by Medicare as liver failure, frequent line sepsis, major central vein thrombosis, or recurrent dehydration. Of these complications, the relationship between liver failure and subsequent death in high-risk subgroups of long-term TPN patients has been shown clearly. Patients with less than 100 cm of postduodenal small bowel, an end-jejunostomy, no ileocecal valve or cecum, or persistently elevated liver function levels are at high risk for end-stage liver disease (ESLD). Early referral to experienced centers is suggested in these circumstances. High-risk patients may also take part in clinical trials of promising therapies to increase intestinal adaptation and prevent liver failure. Living donors should be considered for transplant candidates to minimize waiting time and optimize HLA matching. ESLD patients need a liver-intestine transplant. Because their waiting-list mortality is very high, their status on the liver waiting list should be elevated if possible. High incidence of early death from sepsis is reported after intestinal transplant, even at experienced centers. Aggressive measures should be taken if uncontrolled sepsis occurs, including discontinuing immunosuppression and removing the graft. Further research is needed in intestinal immunology and in development of strategies to decrease the need for aggressive immunosuppression in these transplant recipients. The ultimate role of intestinal transplantation will be determined by its capacity to show superiority, both in effectiveness and safety, to long-term TPN.

Humans↗

Characterization of specific binding sites for vasoactive intestinal peptide in rat intestinal epithelial cell membranes.

Specific binding sites for vasoactive intestinal peptide were characterized in plasma membranes from rat intestinal epithelial cells. At 30 degrees C, the interaction of 125I-labelled peptide with intestinal membranes was rapid, reversible, specific and saturable. At equilibrium, the binding of 125I-labelled peptide was competitively inhibted by native peptide in the 3 . 10(-11)--3 . 10-(7) M range concentration. Scatchard analysis of binding data suggested the presence of two distinct classes of vasoactive intestinal peptide binding sites: a class with a high affinity (Kd = 0.28 nM) and a low capacity (0.8 pmol peptide/mg membrane protein) and a class with a low affininty (Kd = 152 nM) and a high capacity (161 pmol peptide/mg membrane protein). Secretin competitively inhibited binding of 125I-labelled peptide but its potency was 1/1000 that of native peptide. Glucagon and the gastric inhibitory peptide were ineffective. The guanine nucleotides, GTP and Gpp(NH)p inhibited markedly the interaction of 125I-labelled peptide with its binding sites, by increasing the rate of dissociation of peptide bound to membranes. The other nucleotides triphosphate tested (ATP, ITP, UTP, CTP) were also effective in inhibiting binding of 125I-labelled peptide to membranes but their potencies were 1/100--1/1000 that of guanine nucleotides. The specificity and affinity of the vasoactive intestinal peptide-binding sites in plasma membranes prepared from rat intestinal epithelial cells, which is in agreement with an adenylate cyclase highly sensitive to the peptide recently characterized in these membranes (Amiranoff, B., Laburthe, M., Dupont, C. and Rosselin, G. (1978) Biochim. Biophys. Acta 544, 474--481) further argue for a physiological role of the peptide in the regulation of intestinal epithelial function.

Animals↗

Intestinal transplantation in children: differences between isolated intestinal and composite grafts.

The results of the isolated intestinal grafts were compared with those of composite grafts (intestinal graft + liver) in a series of 18 transplantations performed in 17 children; 5 isolated intestinal grafts, 12 hepatointestinal grafts, and 1 multivisceral graft. Causes of intestinal failure were short bowel syndrome (n = 13), motility disorders (n = 2) and congenital epithelial disorders (n = 2). Transplantation was indicated due to end-stage liver disease (n = 14), loss of venous access (n = 2), untreatable diarrhea (n = 1) and high morbidity associated with a poor quality of life (n = 1). Six children, all with a composite graft, died after transplantation due to lymphoma (n = 2), sepsis (n = 1); intraabdominal bleeding (n = 1); pneumonia (n = 1); and overwhelming adenoviral infection (n = 1). Digestive autonomy was achieved in 16 of 18 grafts, the 11 surviving children are free of parenteral nutrition with a reasonably good quality of life. In conclusion, intestinal transplantation is a viable therapeutic alternative for children with permanent intestinal failure. The results of transplantation with an isolated intestine are clearly better that those with a composite graft.

Adolescent↗

Effect of intestinal tapering and lengthening on intestinal structure and function.

BACKGROUND: Intestinal lengthening (TL) is increasingly performed for the short bowel syndrome. Our aim was to evaluate the effect of TL on canine intestinal structure and function. METHODS: Two groups of 5 dogs had 75% distal intestinal resection. Twelve weeks later, serosal electrodes were placed in one group (RO) and tapering and lengthening of 10 to 15 cm distal intestine and electrode placement was performed in the other group (TL). Nutritional status and intestinal absorption, motility, and hormone release were assessed every 4 weeks for 12 weeks after the second procedure. RESULTS: The animals with the lengthening had diminished body weight (75% +/- 2% versus 81% +/- 4%, P < 0.05) and albumin levels (2.4 +/- 0.2 g/dL versus 2.9 +/- 0.5 g/dL, P < 0.05) and impaired absorption (stool fat 18% +/- 6% versus 10% +/- 3%, P < 0.05) compared with those that had undergone resection alone. The intestinal adaptive response was blunted in the TL group (villus height 328 +/- 90 microns versus 410 +/- 36 microns, P < 0.05). Transit time was prolonged 4 and 8 weeks (19 +/- 6 min and 18 +/- 5 min, respectively, versus 11 +/- 3 min, P < 0.05) after TL. Myoelectrical recordings showed a drop in slow wave frequency and impaired migration of Phase III of the migrating motor complex within the tapered and lengthened segments (P < 0.05). Both fasting and postprandial serum gastrin levels were two times greater in the TL group (P < 0.05). Enteroglucagon levels were increased after resection alone; this increase was attenuated by more than 50% in the TL group (P < 0.05). The somatostatin response to feeding was substantially greater in the TL animals. CONCLUSIONS: TL impairs nutritional status and intestinal absorption and adaptation following massive resection. The changes in absorption and transit may be related, in part, to motor disruption and hypergastrinemia. The impaired adaptation is possibly related to decreased enteroglucagon and increased somatostatin levels.

Animals↗

Alkaline phosphatase isozymes in large intestines and large intestinal tumors of Fischer 344 rats.

Eight electrophoretic forms of alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1) were detected in butanol extracts of Fischer 344 rat large intestines. Seven of these isozymes have higher mobility than the small intestinal form(s) in non-denaturing polyacrylamide gel electrophoresis. These more mobile forms may be derived during extraction from more slowly migrating forms that are analogous to those forms found in the small intestine. Monoclonal antibody with specificity for a rat small intestinal isozyme cross-reacts with four of the large intestinal isozymes. This antibody does not cross-react with alkaline phosphatases from human or hog small intestine. Gel exclusion chromatography molecular weight estimates of rat intestinal forms range from 1.1 X 10(5) to 2.5 X 10(5). Alkaline phosphatase from two colon tumors obtained from azoxymethane treated rats appeared to be similar to an isozyme found in some normal rats, on the basis of electrophoretic mobility and cross-reactivity with the monoclonal antibody.

Alkaline Phosphatase↗

A selective inhibitor of intestinal ACAT, EAB309 suppresses both intestinal and hepatic cholesterol output and stimulates chylomicron removal.

The effect of a novel inhibitor of acylcoenzyme A:cholesterol acyltransferase (EC 2.3.1.26, ACAT), EAB309 (EAB) on plasma lipid metabolism was studied in cholesterol-fed rats. Orally administered EAB was not detected in the portal vein or the liver but distributed exclusively in the intestine, suggesting that this agent selectively inhibits intestinal ACAT. The rats were fed with either a cholesterol-diet or a cholesterol-diet containing 0.005% EAB (w/w) ad. libium for three weeks. ACAT activity in intestinal microsomes was significantly inhibited in EAB-treated rats. Hepatic ACAT activity was also decreased in EAB-treated rats, however, this was attenuated by the addition of excess cholesterol to the liver microsome, indicating that substrate availability is tightly associated with this enzyme's activity and the inhibition of hepatic ACAT by EAB is not direct. Incorporation of [3H]-cholesterol to cholesteryl ester (CE) in mesenteric lymph were markedly suppressed by EAB treatment. Chylomicrons (CMs) were doubly labeled with [3H]-vitamin A and [14C]-triglyceride (TG) in EAB-treated or non-treated rats and injected into normal chow-fed rats. The CMs from EAB-treated rats were cleared faster from the plasma and taken up more by the liver compared with the CMs from non-treated rats. The content of CE in newly secreted VLDL was remarkably decreased by EAB treatment without affecting TG output. These results demonstrate that EAB, a novel inhibitor of intestinal ACAT, significantly suppresses both intestinal and hepatic CE output and stimulates CM removal. This suggests that the inhibition of intestinal ACAT can subsequently suppress hepatic ACAT by decreased CE delivery from the intestine to the liver.

Animals↗

A serum factor after intestinal resection stimulates epidermal growth factor receptor signaling and proliferation in intestinal epithelial cells.

BACKGROUND: In vivo, intestinal adaptation after massive small bowel resection (SBR) requires a functional epidermal growth factor (EGF) receptor (EGFR). In vitro studies have shown that serum from mice after SBR induces rat intestinal epithelial cells to proliferate. This study tested the hypothesis that the proliferative response to SBR serum is mediated by EGFR signaling. METHODS: Serum was collected from male Sprague-Dawley rats 7 days after 75% SBR or sham operation. Rat intestinal epithelial cells were incubated in the presence of sham or SBR serum. Total EGFR expression and phosphorylation of several EGFR downstream pathways were determined by Western blotting. In other experiments, a specific EGFR inhibitor (ZD1839) was added and cell growth determined over 5 days. RESULTS: SBR serum significantly increased total EGFR expression (3-fold) over sham operation and consistently activated the phosphatidylinositol 3-kinase pathway. Furthermore, SBR serum markedly augmented rat intestinal epithelial cell growth, an effect that was abolished by EGFR inhibition. CONCLUSIONS: SBR serum contains a factor or factors that stimulates proliferation of intestinal epithelial cells by an EGFR and phosphatidylinositol 3-kinase signaling mechanism. These data recapitulate in vivo studies supporting the hypothesis that EGFR is a central mediator of postresection intestinal adaptation. This in vitro model may provide a novel means to gain insight into the pathophysiology of intestinal adaptation.

Adaptation, Physiological↗

Fasting and postprandial overproduction of intestinally derived lipoproteins in an animal model of insulin resistance. Evidence that chronic fructose feeding in the hamster is accompanied by enhanced intestinal de novo lipogenesis and ApoB48-containing lipoprotein overproduction.

Insulin-resistant states are characterized by hypertriglyceridemia, predominantly because of overproduction of hepatic very low density lipoprotein particles. The additional contribution of intestinal lipoprotein overproduction to the dyslipidemia of insulin-resistant states has not been previously appreciated. Here, we have investigated intestinal lipoprotein production in a fructose-fed hamster model of insulin resistance previously documented to have whole body and hepatic insulin resistance, and hepatic very low density lipoprotein overproduction. Chronic fructose feeding for 3 weeks induced significant oversecretion of apolipoprotein B48 (apoB48)-containing lipoproteins in the fasting state and during steady state fat feeding, based on (a) in vivo Triton WR1339 studies of apoB48 production as well as (b) ex vivo pulse-chase labeling of intestinal enterocytes from fasted and fed hamsters. ApoB48 particle overproduction was accompanied by increased intracellular apoB48 stability, enhanced lipid synthesis, higher abundance of microsomal triglyceride transfer protein mass, and a significant shift toward the secretion of larger chylomicron-like particles. ApoB48 particle overproduction was not observed with short-term fructose feeding or in vitro incubation of enterocytes with fructose. Secretion of intestinal apoB48 and triglyceride was closely linked to intestinal enterocyte de novo lipogenesis, which was up-regulated in fructose-fed hamsters. Inhibition of fatty acid synthesis by cerulenin, a fatty acid synthase inhibitor, resulted in a dose-dependent decrease in intestinal apoB48 secretion. Overall, these findings further suggest that intestinal overproduction of apoB48 lipoproteins should also be considered as a major contributor to the fasting and postprandial dyslipidemia observed in response to chronic fructose feeding and development of an insulin-resistant state.

Acetates↗