Epithelial permeability is not increased in rats following small bowel resection.

BACKGROUND: Increased intestinal permeability and translocation of bacteria and/or bacterial products may cause infection and liver dysfunction in patients with the short bowel syndrome. In previous studies, serum from mice undergoing small bowel resection (SBR) enhanced growth of cultured rat intestinal epithelial cells (RIEC-6), implicating a role for a serum factor(s) in the enterocyte response to SBR. These experiments tested the hypothesis that epithelial cell permeability is increased following SBR. MATERIALS AND METHODS: Male Sprague-Dawley rats underwent a 75% SBR or sham operation. Intestinal permeability in the remnant ileum was determined by Ussing chambers on Postoperative Day (POD) 3. Additionally, serum was collected on POD 1, 3, and 7 and mesenteric lymph was harvested on POD 3. Once confluent, RIEC-6 cells were incubated for 3 days in media supplemented with 10% fetal bovine serum (FBS; control), 1% FBS, 1% FBS plus 9% Sham serum, or 1% FBS plus 9% SBR serum or exposed to media with varied concentrations of SBR or Sham lymph. Monolayer permeability was determined by measuring the passage of dextran-rhodamine. RESULTS: Intestinal permeability was reduced in rats undergoing SBR. Sham serum-treated monolayers demonstrated the greatest permeability. Incubation with SBR serum reduced permeability to near control media. There were no permeability differences between SBR and Sham lymph-treated monolayers. CONCLUSION: The early adaptive response of the remnant intestine after SBR is associated with reduced permeability. These results suggest an alternative mechanism for the increased bacterial translocation that has been described following SBR.

Serum from mice after small bowel resection enhances intestinal epithelial cell growth.

BACKGROUND/PURPOSE: The adaptive response of the intestine to massive small bowel resection (SBR) is remarkably complex. An in vitro model of adaptation may facilitate the elucidation of signaling pathways involved in this process. In an effort to establish such a model, the effects of serum from resected mice on cultured intestinal epithelial cells were studied. METHODS: Serum was collected and pooled from male ICR mice 3 days after either 50% SBR or sham operation. Rat intestinal epithelial cells (RIEC-6) were plated at equal density and grown in the presence of 1% fetal bovine serum (FBS), 10% FBS, 1% FBS plus 9% sham serum, or 1% FBS plus 9% SBR serum. Cell number, proliferation, and caspase-3 activity were determined. RESULTS: RIEC-6 cell growth was reduced significantly in 1% FBS or sham serum. SBR serum markedly accelerated cell growth and proliferation when compared with all other groups and significantly suppressed caspase-3 activity. CONCLUSIONS: Massive intestinal resection in mice results in a serum factor that induces intestinal cell growth in vitro. This in vitro model of trophic signaling will permit further detailed investigations into the mechanisms of intestinal adaptation.

cDNA microarray analysis of adapting bowel after intestinal resection.

BACKGROUND/PURPOSE: Studies of the genetic regulation of various physiologic processes have been hampered by methodologies that are limited to the analysis of individual genes. The advent of cDNA microarray technology has permitted the simultaneous screening of numerous genes for alterations in expression. In this study, cDNA microarrays were used to evaluate gene expression changes during the intestinal adaptive response to massive small bowel resection (SBR). METHODS: Male ICR mice (n = 20) underwent either a 50% SBR or sham operation and then were given either orogastric epidermal growth factor (EGF, 50 microg/kg/d) or saline. After 3 days, cDNA microarray analysis was performed on mRNA extracted from the remnant ileum. RESULTS: From over 8,700 different genes, the array identified 27 genes that were altered 2-fold or greater after SBR. Small proline-rich protein 2 (sprr2), the gene with the greatest expression change (4.9-fold), was further upregulated by EGF. This gene has never been characterized in the intestine or described in intestinal adaptation. CONCLUSIONS: cDNA microarray analysis showed enhanced expression of sprr2, a gene not previously known to be involved in the physiology of adaptation after SBR. This technology provides a more rapid and efficient means of dissecting the complex genetic regulation of gut adaptation.

Analysis of intestinal adaptation gene expression by cDNA expression arrays.

BACKGROUND: As a tool for determining gene expression on a genomic scale, cDNA microarrays are a promising new technology that can be applied to the study of complex physiologic processes. The objective of this study was to characterize the expression of individual genes and patterns of gene expression that might provide insight into the mechanism of intestinal adaptation after massive small bowel resection. METHODS: Male ICR mice underwent a 50% proximal small bowel resection (SBR) or sham operation. After 3 days, the remnant ileum was harvested, weighed, and RNA extracted. Changes in gene expression were detected utilizing Clontech Atlas mouse cDNA expression arrays. Some of these changes were confirmed by reverse transcriptase-polymerase chain reactions (RT-PCR) and Northern blots. RESULTS: Analysis of these cDNA arrays revealed changes in the expression of multiple genes, including those involved in cell cycle regulation, apoptosis, DNA synthesis, and transcriptional regulation. The patterns of expression were consistent with the increased cell proliferation and apoptosis observed during intestinal adaptation. A large number of genes not previously associated with intestinal adaptation were identified. CONCLUSIONS: This technology may facilitate the elucidation of the intricate cellular mechanisms underlying intestinal adaptation.

Epidermal growth factor is critical for intestinal adaptation following small bowel resection.

The loss of small intestinal mucosal surface area is a relatively common clinical situation seen in both the pediatric and adult population. The most frequent causes include mesenteric ischemia, trauma, inflammatory bowel disease, necrotizing enterocolitis, and volvulus. Following surgical resection, the remnant intestine compensates or adapts to the loss of native bowel by increasing its absorptive surface area and functional capacity. Unfortunately, many patients fail to adapt adequately, and are relegated to lifelong intravenous nutrition. Research into intestinal adaptation following small bowel resection (SBR) has evolved only recently from the gross and microscopic level to the biochemical and genetic level. As understanding of this process has increased, numerous therapeutic strategies to augment adaptation have been proposed. Epidermal growth factor (EGF) is an endogenous peptide that is secreted into the gastrointestinal tract and able to influence gut ontogeny, as well as mucosal healing. Early studies have demonstrated its ability to augment the adaptive process. Focusing on a murine model of massive intestinal loss, the morphological, structural, biochemical, and genetic changes that occur during the intestinal adaptive process will be reviewed. The role of EGF and its receptor as critical mediators of the adaptive process will be discussed. Additionally, the ability of EGF to augment intestinal proliferation and diminish programmed cell death (apoptosis) following SBR will be examined. Enhancing adaptation in a controlled manner may allow patients to transition off parenteral nutrition to enteral feeding and, thereby, normalize their lifestyle.

Bax is required for increased enterocyte apoptosis after massive small bowel resection.

BACKGROUND: Massive small bowel resection (SBR) increases rates of both enterocyte proliferation and apoptosis. Previous studies have demonstrated increased intestinal expression of proapoptotic bax mRNA and protein, as well as the appearance of an 18-kd bax cleavage product within 12 hours of SBR. This study tested the hypothesis that bax is required for postresection increases in enterocyte apoptosis. METHODS: Male bax-null and C57Bl/6 (control) mice underwent either a 50% proximal SBR or sham operation. After 3 days, the remnant ileum was harvested and weighed. Apoptotic indexes, proliferation indexes, villus heights, and crypt depths were determined. RESULTS: The usual adaptive increases in ileal wet weight, crypt depth, and rate of proliferation occurred in both the control and bax-null mice. Resection significantly increased the rate of apoptosis in the control mice; however, it failed to alter the apoptotic index in the bax-null mice. CONCLUSIONS: Bax is necessary for the increase in apoptosis that occurs after SBR, but its absence has no significant effect on short-term adaptation. These findings suggest that enterocyte proliferation and apoptosis are differentially regulated during intestinal adaptation.

Salivary epidermal growth factor and intestinal adaptation in male and female mice.

Salivary epidermal growth factor (sEGF) levels are increased in male mice after small bowel resection (SBR) and may be important during intestinal adaptation. Since males have greater sEGF than females, the influence of sex on postresection adaptation was tested. Females had lower sEGF; however, sEGF substantially increased in both sexes after a massive (50%) SBR. Adaptive increases in DNA and protein content, villus height, and crypt depth, as well as crypt cell proliferation rates in the remnant ileum, were not different between males and females. Although significant postresection increases in sEGF were identified, EGF mRNA and protein did not change within the submandibular gland. Glandular kallikrein-13 and ileal EGF receptor expression were greater after SBR in female mice. Intestinal adaptation is equivalent in female and male mice after SBR. Despite lower sEGF, females demonstrated increased expression of a kallikrein responsible for sEGF precursor cleavage as well as amplified ileal EGF receptor expression. These results endorse an important differential response between sexes regarding sEGF mobilization and intestinal receptor availability during adaptation.

Epidermal growth factor alters the bax:bcl-w ratio following massive small bowel resection.

BACKGROUND: Following massive small bowel resection (SBR), the expression of bax and bcl-w is associated with increased enterocyte apoptosis. Epidermal growth factor (EGF) has been shown to enhance enterocyte proliferation and retard apoptosis in the adapting bowel. This study examined the effect of EGF on the expression of these bcl-2 family members during adaptation. MATERIALS AND METHODS: Mice (C57Bl/6; n = 38) underwent a 50% SBR or sham operation and then were randomized to receive twice-daily orogastric saline or EGF (50 microg/kg/day). After 3 days, the remnant ileum was removed, apoptotic index (No. apoptotic bodies/crypt) calculated, and expression of mRNA and protein for bax and bcl-w quantified. RESULTS: EGF prevented the expected increase in the apoptotic index after SBR and altered the ratio of bax to bcl-w in favor of cell survival. CONCLUSION: Following massive small bowel resection, EGF retards rates of enterocyte apoptosis and modifies the expression of bcl-2 family members. By decreasing bax and increasing bcl-w expression, the balance between pro- and anti-apoptotic genes is shifted in favor of cell survival. Alteration of bcl-2 family member expression may be an important mechanism by which EGF reduces the increased enterocyte apoptosis that occurs after massive small bowel resection.

p21 (WAF1/CIP1) is required for the mitogenic response to intestinal resection.

BACKGROUND: Increased enterocyte proliferation and apoptosis characterize the intestinal adaptive response to massive small bowel resection (SBR). Since p21 (WAF1/CIP1) has been implicated to play a role in cellular differentiation and apoptosis, this study tested the hypothesis that p21 is obligatory for adaptation to occur. MATERIALS AND METHODS: p21-null (n = 36) and wild-type (C57B1/6, n = 19) mice underwent a 50% SBR or sham operation. After 3 days, parameters of adaptation (ileal wet weight, villus/crypt morphology, and ileal protein content), an enterocyte proliferation index (PI), and an apoptotic index (AI) were determined in the residual ileum. In a separate set of experiments, p21-null (n = 11) and control (n = 20) mice underwent the aforementioned operative procedures and the remnant intestine was subjected to a reverse transcription polymerase chain reaction for p27 (KIP1). RESULTS: Both AI and PI increased after SBR in the wild-type mice. In the p21-null mice, SBR increased AI, but did not affect the PI. After SBR, adaptive parameters increased in the wild-type mice, but failed to increase in the p21-null mice. The absence of p21 caused a baseline increase in p27 mRNA, which did not change after SBR. CONCLUSION: p21 appears to be required to increase enterocyte proliferation and to augment the other parameters of intestinal adaptation. In the absence of p21, the proliferative and apoptotic responses to SBR are uncoupled. These results suggest a differential mechanism for the regulation of enterocyte proliferation and apoptosis in the adapting intestine.

Intestinal adaptation occurs independent of transforming growth factor-alpha.

BACKGROUND/PURPOSE: Signal transduction via the epidermal growth factor receptor (EGFR) is critical for intestinal adaptation after massive small bowel resection (SBR). Although it has been assumed that the major ligand for the EGFR during adaptation is EGF, the role for transforming growth factor-alpha (TGF-alpha), another major ligand for the EGFR is unknown. The purpose of this study was to test the hypothesis that TGF-alpha is an important ligand for the EGFR during intestinal adaptation. METHODS: Wild-type mice (C57BI/6) underwent a 50% proximal SBR or sham operation (bowel transection or reanastomosis) and were then assigned randomly to receive either intraperitoneal TGF-alpha or placebo. In a separate experiment, SBR or sham operations were performed in mice lacking TGF-alpha (Waved-1). After 3 days, adaptation was measured in the ileum. RESULTS: Exogenous TGF-alpha enhanced intestinal adaptation in the wild-type mice after SBR as shown by increased ileal wet weight and DNA content. Normal adaptation occurred in the mice lacking TGF-alpha as shown by increased ileal wet weight, protein and DNA content, proliferation, villus height, and crypt depth. CONCLUSIONS: Although exogenous TGF-alpha enhanced adaptation after massive SBR, adaptation was preserved in TGF-alpha-absent mice. These results refute TGF-alpha as an essential ligand for EGFR signaling during intestinal adaptation.

Diminished epidermal growth factor levels in infants with necrotizing enterocolitis.

BACKGROUND/PURPOSE: Because epidermal growth factor (EGF) is trophic to the intestinal mucosa, and neonatal necrotizing enterocolitis (NEC) is associated with a disrupted intestinal mucosal barrier, the authors sought to determine whether diminished levels of EGF were present in infants with NEC. METHODS: Saliva, serum, and urine specimens were obtained from infants with NEC during a 3-year period (February 1995 to May 1998). Control patients without NEC were chosen based on similar postnatal age and birthweight. EGF levels were determined by enzyme-linked immunosorbent assay (ELISA). Differences between groups were compared using Mann-Whitney Rank sum test with P less than .05 considered significant. Results are presented as mean values +/-SEM. RESULTS: Twenty-five infants with NEC were compared with 19 control patients. Birth weight (1,616+/-238 g control v. 1,271+/-124 g NEC) and postnatal age (23+/-6 days control v. 22+/-3 days NEC) were similar. Infants with NEC had significantly lower levels of EGF in both saliva (590+/-80 pg/mL control v. 239+/-41 pg/mL NEC; P<.001) and serum (35+/-8 pg/mL control v. 5.6+/-1.9 pg/mL NEC; P<.001). Urinary EGF was also lower in the NEC group, but was not statistically significant. CONCLUSIONS: Premature infants with NEC have significantly diminished levels of salivary and serum EGF. Reduced levels of this growth factor may distinguish infants at risk for NEC and play a pivotal role in the pathogenesis of the perturbed intestinal mucosal barrier that is central to this condition.

Effect of massive small bowel resection on the Bax/Bcl-w ratio and enterocyte apoptosis.

Following small bowel resection (SBR), the remnant intestine undergoes adaptation. Enterocyte proliferation is increased and counterbalanced by increased rates of apoptosis. To elucidate a mechanism for increased enterocyte apoptosis, this study tested the hypothesis that the ratio between pro-apoptotic Bax and pro-survival Bcl-w correlates with the apoptosis that occurs following SBR. Mice (C57Bl/6; n = 76) underwent a 50% proximal SBR or sham operation. After 12 hours and 1, 2, 3, and 7 days, the ileum was removed, the apoptotic index (apoptotic bodies/crypt) was recorded, and the messenger RNA and protein for Bax and Bcl-w were quantified. The apoptotic index was equivalent in the sham and SBR mice at 12 hours; however, it was significantly elevated following SBR at every other day measured. The ratio of Bax to Bcl-w messenger RNA relative to sham operation increased after SBR at 24 hours, decreased by day 3, and returned to baseline levels by 1 week. The protein ratio showed an increase by day 1, which remained elevated through day 7. An augmented ratio of Bax to Bcl-w messenger RNA and protein corresponded with the increase in enterocyte apoptosis. Alterations in the expression ratio of these genes may play a role in establishing a new homeostatic set point between proliferation and apoptosis during adaptation.

Differential expression of ileal Na(+)/H(+) exchanger isoforms after enterectomy.

BACKGROUND: Na(+)/H(+) exchangers (NHE) are transporters involved in the absorption of NaCl along the gastrointestinal tract. The aim of this study was to determine the expression pattern of the intestinal brush border NHE isoforms 2 and 3 following massive small bowel resection (SBR). Additionally, the effect of epidermal growth factor (EGF) and salivarectomy (removal of the primary source of EGF) on the expression pattern was studied. MATERIALS AND METHODS: ICR mice underwent a proximal SBR or sham surgery and then received either orogastric saline or EGF (50 microg/kg/day). In separate experiments mice underwent salivarectomy followed by SBR or sham. Postoperatively the remaining ileum was isolated and levels of NHE-2 and NHE-3 mRNA and protein were resolved. RESULTS: Following SBR, the expression of both mRNA and protein for NHE-3 increased by approximately 2.5-fold. Treatment with EGF enhanced NHE-3 mRNA in sham animals with further elevation following SBR. The expression of NHE-2 mRNA demonstrated minimal change while protein marginally increased (40%) following SBR. EGF did not affect the expression of NHE-2 mRNA. Salivarectomy did not influence NHE-2 protein expression and inhibited the increased NHE-3 protein expression following SBR. CONCLUSIONS: Following SBR, the expression pattern for brush border NHE isoforms is distinctive. Increased expression of NHE-3 secondary to SBR and/or EGF treatment with loss of this increase following salivarectomy implies a common mechanism to enhance enterocyte proliferation and luminal absorption of NaCl and water. These results suggest that NHE-3 is an important ileal exchanger following SBR.

Intestinal overexpression of EGF in transgenic mice enhances adaptation after small bowel resection.

The effect of direct intestinal overexpression of epidermal growth factor (EGF) on postresection adaptation has been investigated by the production of transgenic mouse lines. A murine pro-EGF cDNA construct was produced, and expression of the EGF construct was targeted to the small intestine with the use of the rat intestinal fatty acid-binding protein promoter. An approximately twofold increase in intestinal EGF mRNA and protein was detected in heterozygous mice. No changes in serum EGF levels were noted. Except for a slightly shortened small intestine, no other abnormal phenotype was observed. Intestinal adaptation (increases in body weight, DNA, protein content, villus height, and crypt depth) was markedly enhanced after a 50% proximal small bowel resection in transgenic mice compared with nontransgenic littermates. This transgenic mouse model permits the study of intestinal adaptation and other effects of EGF in the small intestine in a more physiological and directed manner than has been previously possible. These results endorse a direct autocrine/paracrine mechanism for EGF on enterocytes as a means to enhance adaptation.

Intestinal adaptation and enterocyte apoptosis following small bowel resection is p53 independent.

Adaptation following small bowel resection (SBR) signals enterocyte proliferation and apoptosis. Because p53-induced p21(waf1/cip1) may be important for apoptosis in many cells, we hypothesized that these genes are required for increased enterocyte apoptosis during adaptation. Male C57BL/6 (wild-type) or p53-null mice underwent 50% proximal SBR or sham operation (bowel transection-reanastomosis). Adaptation (DNA-protein content, villus height-crypt depth, enterocyte proliferation), appearance of apoptotic bodies, and p53 and p21(waf1/cip1) protein expression were measured in the ileum after 5 days. Adaptation was equivalent after SBR in both wild-type and p53-null mice as monitored by significantly increased ileal DNA-protein content, villus height, and enterocyte proliferation. The number of crypt apoptotic bodies increased significantly after SBR evenly in both wild-type and p53-null mice. In the p53-null mice, SBR substantially induced the expression of p21(waf1/cip1) protein in villus enterocytes. The p53-independent induction of p21(waf1/cip1) may account for the similar intestinal response to SBR between wild-type and p53-null mice. Intestinal adaptation and increased enterocyte apoptosis following intestinal resection occur via a p53-independent mechanism.

The expression and activation of EGF and c-NEU receptors are increased in enterocytes during intestinal adaptation.

BACKGROUND/PURPOSE: After massive small bowel resection (SBR), epidermal growth factor (EGF) and its intestinal receptor (EGF-R) play major roles during adaptation. The expression of a homologous enterocyte receptor termed c-neu (c-neu-R) is capable of forming heterodimers with EGF-R to facilitate cellular signaling. The purpose of this study was to determine the expression and activation of EGF-R and c-neu-R during the adaptive intestinal response to SBR. METHODS: Male ICR mice underwent either SBR or sham surgery. After 1, 3, and 7 days, enterocytes were isolated and protein immunoprecipitated with antibody to either EGF-R or c-neu-R. Receptor protein expression and activation status were determined. RESULTS: When compared with sham operation, the expression and activation status of both EGF-R (six- and twofold, respectively) and c-neu-R (nine- and twofold, respectively) were increased substantially in enterocytes from the adapting ileum after SBR by postoperative day 3. Minimal changes were appreciated for either EGF-R or c-neu-R expression or activation in the remnant bowel after enterocyte removal, liver, or kidney. CONCLUSIONS: Both the expression and activation status of EGF-R and c-neu-R are increased substantially in enterocytes from the adapting ileum by postoperative day 3 after massive SBR. These changes provide a unique mechanism for the enterocyte to enhance cellular signaling in response to EGF during intestinal adaptation.

The adaptive intestinal response to massive enterectomy is preserved in c-SRC-deficient mice.

BACKGROUND/PURPOSE: The Src family of protein tyrosine kinases has been implicated in the downstream mitogenic signaling of several ligands including epidermal growth factor (EGF). Because EGF likely plays a role in adaptation after massive small bowel resection (SBR), we tested the hypothesis that c-src is required for this important response. METHODS: A 50% proximal SBR or sham operation (bowel transection or reanastomosis alone) was performed on c-src-deficient (n = 14) or wild-type (C57bl/6) mice (n = 20). The ileum was harvested on postoperative day 3 and adaptive parameters determined as changes in ileal wet weight, protein and DNA content, proliferation index, villus height, and crypt depth. Comparisons were done using analysis of variance (ANOVA), and a Pvalue less than .05 was considered significant. Values are presented as mean +/- SEM. RESULTS: The activity of c-src was increased in the ileum of wild-type mice after SBR but remained unchanged in c-src-deficient mice. Despite this lack of increase, adaptation occurred after SBR in the c-src-deficient mice as demonstrated by increased ileal wet weight, protein and DNA content, proliferation index, villus height, and crypt depth similar to wild-type mice. CONCLUSIONS: The adaptive response of the intestine to massive SBR is preserved despite reduced activity of the c-src protein. The mitogenic signaling that characterizes intestinal adaptation and is associated with receptor activation by EGF or other growth factors probably occurs by mechanisms independent of c-src protein tyrosine kinase.

Apoptosis and the pattern of DNase I expression following massive small bowel resection.

INTRODUCTION: Following massive small bowel resection (SBR), histologic evidence of increased enterocyte apoptosis has been demonstrated in several animal models. Deoxyribonuclease I (DNase I) is requisite for intranuclear cleavage of DNA during apoptosis; we therefore hypothesized that the expression of this gene would be increased following SBR. METHODS: Male ICR mice underwent either 50% proximal SBR or sham surgery (bowel transection/reanastomosis). After 12 h and 1, 3, and 7 days, rates of enterocyte proliferation and apoptosis were recorded as well as DNase I mRNA expression and activity. RESULTS: Adaptation after SBR was confirmed at each time point by increased proliferation. Enterocyte proliferation was increased by 12 h and apoptosis was increased by 24 h and remained elevated through Day 7. When compared with sham-operated mice, SBR resulted in a twofold increase in both DNase I expression and activity at 24 h postoperatively, which returned to baseline by Postoperative Day 3. CONCLUSIONS: DNase I expression and activity are increased early following massive SBR but return to baseline despite persistent increased rates of enterocyte apoptosis and proliferation. This enzyme may be important in the early induction of apoptosis following massive SBR, but not once a new set point has been established in the balance between the rate of enterocyte production and enterocyte loss.

The distribution of endogenous epidermal growth factor after small bowel resection suggests increased intestinal utilization during adaptation.

BACKGROUND/PURPOSE: Although exogenous epidermal growth factor (EGF) amplifies adaptation after massive small bowel resection (SBR), the role for endogenous EGF is unclear. The authors sought to determine whether SBR was associated with changes in the levels of EGF in the serum, saliva, or urine and EGF receptor (EGF-R) signaling in the ileum. METHODS: Male ICR mice underwent 50% proximal SBR or sham surgery bowel transection/reanastomosis). After 3 days, levels of EGF were measured by enzyme-linked immunosorbent assay (ELISA) in the serum, saliva, and urine. EGF-R activation was measured in isolated ileal enterocytes by probing an EGF-R immunoprecipitate with an antibody to phosphotyrosine. RESULTS: When compared with sham, SBR resulted in no change in serum, increased salivary (2209+/-266 nmol SBR v 1183+/-119 nmol sham, P<.05) and decreased urinary (417+/-58 nmol SBR v 940+/-143 nmol sham; P<.05) EGF levels. EGF-R activation increased 2.5-fold after SBR. CONCLUSIONS: Increased salivary and reduced urinary EGF linked with enhanced EGF-R activation suggests increased ileal utilization of EGF during adaptation. This observation, along with the known beneficial effects of exogenous EGF, infers a crucial role for endogenous EGF in the pathogenesis of intestinal adaptation after SBR.

The effect of epidermal growth factor on differentiation of isolated enterocytes after small bowel resection.

BACKGROUND/PURPOSE: In previous studies using mucosal scrapings or whole-bowel homogenates, epidermal growth factor (EGF) augments adaptation after massive small bowel resection (SBR). The purpose of this study was to determine directly the effect of adaptation and EGF on enterocyte differentiation using an explicit enterocyte cell population. METHODS: Male ICR mice underwent 50% proximal SBR or sham (bowel transection-reanastomosis) and were selected randomly to either orogastric saline or EGF (50 microg/kg/d). After 3 days, enterocytes were isolated from the remnant ileum by mechanical vibration and assayed for DNA and protein content as well as sucrase and alkaline phosphatase (AlkP) activity. RESULTS: Ileal wet weight, enterocyte protein, and DNA content were increased significantly after SBR and boosted even further with EGF. When normalized for protein, SBR caused an increase in AlkP and sucrase activity, and EGF treatment caused AlkP and sucrase activity to return to baseline. CONCLUSIONS: EGF enhances adaptation; however, when normalized for protein, the activity of two enterocyte-specific enzymes was not significantly altered by EGF. This analysis of an explicit enterocyte population supports the notion that the beneficial effects of EGF are more likely caused by increased numbers of enterocytes rather than an increase in the functional activity of each individual cell.