Open Access

Enteroendocrine cells, stem cells and differentiation progenitors in rats with TNBS-induced colitis

  • Authors:
    • Magdy El-Salhy
    • Tarek Mazzawi
    • Kazuo Umezawa
    • Odd Helge Gilja
  • View Affiliations

  • Published online on: October 24, 2016     https://doi.org/10.3892/ijmm.2016.2787
  • Pages: 1743-1751
  • Copyright: © El-Salhy et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Patients with inflammatory bowel disease (IBD), as well as animal models of human IBD have abnormal enteroendocrine cells. The present study aimed to identify the possible mechanisms underlying these abnormalities. For this purpose, 40 male Wistar rats were divided into 4 groups as follows: the control group, the group with trinitrobenzene sulfonic acid (TNBS)-induced colitis with no treatment (TNBS group), the group with TNBS-induced colitis treated with 3-[(dodecylthiocarbonyl)-methyl]-glutarimide (DTCM-G; an activator protein-1 inhibitor) (DTCM-G group), and the group with TNBS-induced colitis treated with dehydroxymethylepoxyquinomicin (DHMEQ; a nuclear factor-κB inhibitor) treatment (DHMEQ group). Three days following the administration of TNBS, the rats were treated as follows: those in the control and TNBS groups received 0.5 ml of the vehicle [0.5% carboxymethyl cellulose (CMC)], those in the DTCM-G group received DTCM-G at 20 mg/kg body weight in 0.5% CMC, and those in the DHMEQ group received DHMEQ at 15 mg/kg body weight in 0.5% CMC. All injections were administered intraperitoneally twice daily for 5 days. The rats were then sacrificed, and tissue samples were taken from the colon. The tissue sections were stained with hemotoxylin-eosin and immunostained for chromogranin A (CgA), serotonin, peptide YY (PYY), oxyntomodulin, pancreatic polypeptide (PP), somatostatin, Musashi1 (Msi1), Math1, Neurogenin3 (Neurog3) and NeuroD1. The staining was quantified using image analysis software. The densities of CgA-, PYY-, PP-, Msi1-, Neurog3- and NeuroD1-positive cells were significantly lower in the TNBS group than those in the control group, while those of serotonin-, oxyntomodulin- and somatostatin-positive cells were significantly higher in the TNBS group than those in the control group. Treatment with either DTCM-G or DHMEQ restored the densities of enteroendocrine cells, stem cells and their progenitors to normal levels. It was thus concluded that the abnormalities in enteroendocrine cells and stem cells and their differentiation progenitors may be caused by certain signaling substances produced under inflammatory processes, resulting in changes in hormone expression in enteroendocrine cells. These substances may also interfere with the colonogenic activity and the differentiation of the stem-cell secretory lineage into mature enteroendocrine cells.

Introduction

The large intestine contains five types of enteroendocrine cells: namely serotonin-, polypeptide YY (PYY)-, oxyntomodulin (enteroglucagon)-, pancreatic polypeptide (PP)- and somatostatin-producing cells (1). In addition, chromogranin A (CgA) is expressed by all enteroendocrine cells and is used as a common marker for them (2). The interaction between enteroendocrine and immune cells during inflammation was recently discussed, and this interaction is thought to play a pivotal role in the inflammatory process (3).

Patients with inflammatory bowel disease (IBD), as well as animal models of human IBD have been shown to have abnormal enteroendocrine cells (424). The nature of the changes in enteroendocrine cells differs between ulcerative colitis (UC), Crohn's disease (CD) and microscopic colitis (423). The mechanisms underlying such abnormalities are not yet known. However, a recent study using an animal model of human UC, namely dextran sulfate sodium-induced colitis, found that the abnormalities in enteroendocrine cells strongly correlated with the abnormal differentiation progeny of stem cells (25). It has been suggested that the abnormalities in the enteroendocrine cells in this animal model are caused by an abnormal stem cell differentiation progeny toward enteroendocrine cells (25).

Trinitrobenzene sulfonic acid (TNBS)-induced colitis in experimental animals is commonly used as an animal model of human CD (26). The enteroendocrine cells in this animal model have been reported to be abnormal (27). The treatment colitis with activator protein 1 (AP-1) and nuclear factor-κB inhibitors, which are potent anti-inflammatory agents, has been shown to restore enteroendocrine cells to normal levels (27).

The aim of the present study was to determine whether the changes in the densities of enteroendocrine cells in TNBS-induced colitis involve stem cell differentiation and/or the cellular expression of enteroendocrine cell hormones.

Materials and methods

Rats

A total of 40 male Wistar rats (Hannover GALAS; Taconic Farms Inc., Lille Skensved, Denmark) with a mean body weight of 200 g (range, 160–250 g) were housed in Macrolon III cages with water and food available ad libitum. They were fed a standard diet (B&K Universal, Nittedal, Norway) and were kept at a temperature of 18–22°C, a relative humidity of 50–60%, and under a 12/12-h light/dark cycle. The rats were allowed to acclimatize to the conditions in the animal house for at least 7 days prior to being used in the experiments. The rats were divided into the following 4 groups containing 10 animals in each: i) the control group; ii) the group with TNBS-induced colitis with no treatment (TNBS group); iii) the group with TNBS-induced colitis treated with 3-[(dodecylthiocarbonyl)-methyl]-glutarimide (DTCM-G; an activator protein-1 inhibitor) (DTCM-G group); and iv) the group with TNBS-induced colitis treated with dehydroxymethylepoxyquinomicin (DHMEQ; a nuclear factor-κB inhibitor) (DHMEQ group).

This study was performed in accordance with the Directive for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes (86/609/EEC), in compliance with the Helsinki Declaration. The Local Ethics Committee for Experimental Animals at the University of Bergen (Bergen, Norway) approved the study.

Use of TNBS to induce colitis

Colitis was induced in the rats in the TNBS, DTCM-G and DHMEQ groups as previously described (28) using a single dose of TNBS (Sigma-Aldrich Logistik, Steinheim, Germany). The animals were anesthetized with isoflurane (Schering-Plough Pharmaceuticals, North Wales, PA, USA), and TNBS was administered into the colon at 8 cm from the anal margin (25 mg/animal in 50% ethanol solution; 0.5 ml/rat), followed by 2 ml of air. TNBS was administered via an 8.5-cm-long, 2.5-mm-diameter round-tipped Teflon feeding tube (AngTheo, Lidingö, Sweden). The animals were kept in the prone position with their hind legs raised for approximately 3 min following the administration of TNBS. They were supervised until recovery and then monitored daily. The rats in the control group were subjected to the same procedure as the rats in the TNBS group, except that 0.9% saline was introduced into the colon instead of TNBS.

Treatment with DTCM-G and DHMEQ

Three days following the administration of TNBS, the rats were treated as follows: those in the control and TNBS groups received 0.5 ml of the vehicle [0.5% carboxymethyl cellulose (CMC)], those in the DTCM-G group received DTCM-G at 20 mg/kg body weight in 0.5% CMC, and those in the DHMEQ group received DHMEQ at 15 mg/kg body weight in 0.5% CMC. All injections were administered intraperitoneally twice daily for 5 days. The synthesis of DTCM-G and DHMEQ is described in detail elsewhere (2934). Animals exhibiting signs of pain were administered a subcutaneous injection of 1 ml of a 0.3-g/ml Temgesic solution (Merck Pharmaceuticals, Kenilworth, NJ, USA). At the end of the experiments, the animals were sacrificed by the inhalation of CO2 and tissue samples were obtained from the colon.

Histopathological and immunohistochemical examinations

The colonic tissues were fixed in 4% buffered paraformaldehyde overnight, embedded in paraffin, and cut into 5-μm-thick sections, which were stained with hematoxylin and eosin (ThermoFischer Scientific, Waltham. MA, USA). Inflammation was evaluated using the scoring system of Hunter et al (35). The sections were also immunostained using the ultraView Universal DAB Detection kit (version 1.02.0018) and the BenchMark Ultra IHC/ISH staining module (both from Venata Medical Systems, Basel, Switzerland). The sections were incubated with the primary antibodies for 32 min at 37°C. Details of the primary antibodies used are presented in Table I.

Table I

Primary antibodies used in immunohistochemical staining.

Table I

Primary antibodies used in immunohistochemical staining.

Antibodies raised againstSourceCode no.Working dilutionType of antibodyDetects
N-terminal of purified CgADako (Glostrup, Denmark)M8691:1,000Monoclonal, raised in mouseCgA
SerotoninDako5HT-2091:1,500Monoclonal, raised in mouseSerotonin
PYYAlpha-Diagnostica (San Antonio, TX, USA)PYY 11A1:1,000Polyclonal, raised in rabbitPYY
Porcine glicentin/glucagonAcris antibodies (Herford, Germany)BP5081:800Polyclonal, raised in rabbitOxyntomodulin (enteroglucagon)
Synthetic human PPDiagnostic Biosystems (Pleasanton, CA, USA)#1141:400Polyclonal, raised in rabbitPP
Synthetic human somatostatinDakoA5661:200Polyclonal, raised in rabbitSomatostatin
Residues 5-21 [APQP GLASPDSPHDPCK] of the human, mouse and rat Msi1Novus Biologicals Europe (Abingdon, UK)NB100-17591:100Polyclonal, raised in rabbitMsi1
Sy\nthetic peptide surrounding amino acid 190 of human Math1BioVision (Milpitas, CA, USA)3658-1001:50Polyclonal, raised in rabbitMath1
KLH-conjugated synthetic peptide between 40–69 amino acids from the N-terminal region of human Neurog3ThermoFisher Scientific (Oslo, Norway)BT-B561801:50Polyclonal, raised in rabbitNeurog3
Recombinant full-length human NeuroD1Nordic BioSite (Täby, Sweden)PA5-118931:100Polyclonal, raised in rabbitNeuroD1

[i] CgA, chromogranin A; Msi, Musashi1; Neurog3, Neurogenin3; PYY, peptide YY; PP, pancreatic polypeptide.

Morphometry

The endocrine cells were quantified using image analysis software (version 1.7, cellSens; Olympus, Tokyo, Japan). The numbers of endocrine, and Musashi (Msi1)-, Math1-, Neurogenin3 (Neurog3)- and NeuroD1-positive cells were counted manually. The area of epithelial cells was determined manually by drawing an enclosed region using the computer mouse.

The densities of endocrine cells were expressed as the number of immunoreactive endocrine cells per square millimeter of epithelium, the density of Msi1 cells was expressed as the number of immunoreactive cells per crypt, and the densities of Math1, Neurog3 and NeuroD1 cells were expressed as the number of immunoreactive cells per field. Quantification was performed in 10 randomly chosen microscopic fields using a ×40 objective. The measurements were made by the same individual (M.E.-S.) who was blind to the identities of the slides.

Statistical analysis

Differences between the control, TNBS, DTCM-G and DHMEQ groups were analyzed using the Kruskal-Wallis non-parametric test, with Dunn's test as a post-test. The correlations between abnormalities in the densities of PYY/oxyntomodulin-, CgA/serotonin- and PP/somatostatin-positive cells were determined using the non-parametric Spearman correlation test. The data are presented as the mean ± SEM values. Probability values of P<0.05 were considered to indicate statistically significant differences.

Results

Two animals died spontaneously in the TNBS group. There were no deaths in the other 3 groups.

Histopathological and immunohistochemical examinations

The histopathological inflammation scores were 6.4±1.1, 1.8±1.2 and 2.3±0.9 in the TNBS, DTCM-G and DHMEQ groups, respectively (Kruskal-Wallis test, P=0.002). Dunn's test showed that the scores differed between the TNBS group, and the DTCM-G and DHMEQ groups (P=0.04 and 0.02, respectively) (data not shown).

CgA-, serotonin-, PYY-, oxyntomodulin-, PP-, somatostatin-, Msi1-, Math1-, Neurog3- and NeuroD1-positive cells were found in all the colonic tissues from the rats in all groups. The CgA-, serotonin-, PYY-, oxyntomodulin-, PP- and somatostatin-positive cells were located mostly in the crypts of Lieberkühn. Msi1-positive cells were found exclusively in the crypts of Lieberkühn. Msi1-positive cells in rats with colitis tended to accumulate at the margins of deep ulcers. Math1-, Neurog3- and NeuroD1-positive cells were observed in the crypts and alongside the gland of Lieberkühn (Figs. 2 and 6).

Morphometry

The results of the quantification of the different types of endocrine cells, stem cells and differentiation progenitors in all 4 experimental groups are summarized in Tables II and III.

Table II

Densities of colonic enteroendocrine cells in the 4 experimental groups.

Table II

Densities of colonic enteroendocrine cells in the 4 experimental groups.

Endocrine cell typeControlsTNBSDTCM-GDHMEQ
CgA-positive111.8±17.951.0±21.1a104.0±14.6107.6±17.1
Serotonin-positive40.6±6.662.6±7.5b39.6±6.237.0±5.4
PYY-positive87.3±2.714.1±3.0c83.8±4.083.9±2.6
Oxyntomodulin-positive44.8±3.778.3±6.8c48.3±4.449.3±3.8
PP-positive58.0±3.531.8±7.5c69.3±6.260.4±4.4
Somatostatin-positive43.6±3.269.9±7.8b40.5±3.243.7±5.1

{ label (or @symbol) needed for fn[@id='tfn2-ijmm-38-06-1743'] } Data are the mean ± SEM values. TNBS, trinitrobenzene sulfonic acid; DTCM-G, 3-[(dodecylthiocarbonyl)-methyl]-glutarimide; DHMEQ, dehydroxymethylepoxyquinomicin; PP, pancreatic polypeptide; PYY, peptide YY; CgA, chromogranin A.

a P<0.05,

b P<0.01

c P<0.001 vs. controls.

Table III

Densities of colonic stem cells and differentiation progenitors in the 4 experimental groups.

Table III

Densities of colonic stem cells and differentiation progenitors in the 4 experimental groups.

Cell typeControlsTNBSDTCM-GDHMEQ
Msi1-positive4.8±0.51.9±0.3b4.9±0.54.5±0.5
Math1-positive72.2±8.598.8±12.797.2±9.398.3±12.7
Neurog3-positive70.9±11.243.6±3.8a106.0±12.679.5±12.0
NeuroD1-positive68.8±10.444.5±7.2a107.5±11.781.0±12.4

{ label (or @symbol) needed for fn[@id='tfn6-ijmm-38-06-1743'] } Data are the mean ± SEM values. TNBS, trinitrobenzene sulfonic acid; DTCM-G, 3-[(dodecylthiocarbonyl)-methyl]-glutarimide; DHMEQ, dehydroxymethylepoxyquinomicin; Msi1, Musashi1; Neurog3, Neurogenin3.

a P<0.05

b P<0.01 vs. controls.

The Kruskal-Wallis test showed that there were significant differences between the experimental groups regarding both PYY- and oxyntomodulin-positive cells (P=0.0003 and 0.001, respectively). Whereas the density of PYY-positive cells was significantly reduced in the TNBS group relative to the controls, the density of oxyntomodulin-positive cells was significantly increased (P<0.0001 for in both) (Figs. 1 and 2). The density of PYY-positive cells inversely correlated with the density of oxyntomodulin-positive cells (r=−0.7, P=0.04).

The densities of CgA- and serotonin-positive cells differed significantly between the control, TNBS, DTCM-G and DHMEQ groups (P=0.04 and 0.006, respectively). In the TNBS group, the density of CgA-positive cells was significantly reduced (P=0.02) and that of serotonin-positive cells was increased (P=0.004) (Fig. 3). The density of CgA-positive cells inversely correlated with the density of serotonin-positive cells (r=−0.7, P=0.03).

The Kruskal-Wallis test showed that there were significant differences in both the PP-positive and somatostatin-positive cell densities between the control and experimental groups (P= 0.002 and 0.01, respectively). While the density of PP-positive cells was reduced in the TNBS group relative to controls (P=0.001) (Fig. 4), that of somatostatin-positive cells was increased (P=0.006). The density of PP-positive cells inversely correlated with the density of serotonin-positive cells (r=−0.8, P=0.004).

The Kruskal-Wallis test showed that there were signifi-cant differences in the densities of Msi1-, Neurog3- and NeuroD1-positive cells, but not in those of Math1-positive cells (P=0.0008, 0.006, 0.003 and 0.2, respectively). The densities of Msi1-, Neurog3- and NeuroD1-positive cells were reduced relative to the controls (P=0.0004, 0.04 and 0.03, respectively), whereas the density of Math1-positive cells was not (P=0.1) (Figs. 5 and 6).

Discussion

The interaction between enteroendocrine cells and immune cells has been recently debated, and it is believed that such an interaction plays an important role in the pathophysiology of IBD (3,3640). Enteroendocrine cells in the same animal model for human CD studied herein have previously been reported to be abnormal (27). The mechanisms underlying these abnormalities however, are unknown.

It is well known that two hormones can be localized in the same enteroendocrine cell, namely glucagon-like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP) in the small intestine, and PYY and oxyntomodulin in the distal small and large intestines (4144). Recent studies have further demonstrated that mature enteroendocrine cells are capable of expressing up to 7 different hormones (4547). In the present study, the increase in the oxyntomodulin-positive cell density was accompanied by a decrease in the PYY-positive cell density, with a significant inverse correlation. Similar observations were found concerning the CgA/serotonin-positive and PP/somatostatin-positive cell densities. It is reasonable to assume that the inflammatory process affects enteroendocrine cells so that they 'switch off' the expression of a certain hormone and 'switch on' the expression of another.

The intestine contains 4 to 6 stem cells per crypt, which either divide into new stem cells (self-renewal; clonogeny) or differentiate into all types of epithelial cells (differentiation progeny) (4859). The differentiation progeny includes two lineages: secretory and absorptive. The secretory lineage gives rise to goblet, endocrine and Paneth cells, while the absorptive lineage gives rise to absorptive enterocytes (4859). Msi1 is a transcription factor expressed by both intestinal stem cells and their early progeny (6063). Math1 is expressed in the secretory lineage by an early progenitor, and mutant (Math1−/−) mice have no secretory cells (64). Neurog3 is expressed by an early progenitor in the secretory lineage, which directs the differentiation of secretory progenitors into endocrine cells (65). Transgenic mice (Neurog3−/−) express normal densities of goblet and Paneth cells, but no enteroendocrine cells at all (6567). NeuroD1 is expressed by progenitors derived from Neurog3 progenitors (68,69). Mice deficient in NeuroD1 do not have a certain subgroup of enteroendocrine cells (66,70).

In this study, the density of Msi1-immunoreactive cells in TNBS-induced colitis was reduced relative to the controls, indicating that the clonogenic activity of the stem cells is affected by inflammation. On the other hand, the density of Math1-immunoreactive cells did not differ between the group with TNBS-induced colitis and the controls, suggesting that inflammation does not interfere with the early secretory lineage differentiation. The present observation that the densities of both Neurog3- and NeuroD1-positive cells were lower in rats with TNBS-induced colitis than in the controls may indicate a decease in the differentiation of stem cells into enteroendocrine cells.

The reduction in enteroendocrine cells observed in this study following the induction of colitis by TNBS seems to be caused by i) the 'switching on' and 'switching off' of the expression of certain hormones by enteroendocrine cells, and ii) decreases in the clonogenic activity of the stem cell and in the differentiation into enteroendocrine cells from stem cell progenitors. It may be speculated that the inflammatory processes trigger certain signaling substances that cause certain enteroendocrine cells to change their hormone expression. These substances may also affect the colonogenic activity and the differentiation of the stem cell secretory lineage into mature enteroendocrine cells.

The 'switching on and off' of the expression of hormones of enteroendocrine cells must occur on a timescale of minutes or hours, and stem cells differentiate into mature intestinal cells in 2–3 days (61). This explains why changes in the densities of enteroendocrine cells, stem cells and differentiation progeny to enteroendocrine cells could be detected 3 days after the induction of colitis using TNBS, and that the treatment of colitis for 5 days with anti-inflammatory agents restored their densities to normal levels.

Acknowledgments

This study was supported by grants from Helse-Vest (grant no. 911978) and Helse-Fonna (grant no. 40515).

References

1 

El-Salhy M, Mazzawi T, Hausken T and Hatlebakk JG: Interaction between diet and gastrointestinal endocrine cells. Biomed Rep. 4:651–656. 2016.PubMed/NCBI

2 

El-Salhy M, Gilja OH, Gundersen D, Hatlebakk JG and Hausken T: Duodenal chromogranin a cell density as a biomarker for the diagnosis of irritable bowel syndrome. Gastroenterol Res Pract. 2014:4628562014. View Article : Google Scholar : PubMed/NCBI

3 

El-Salhy M and Hausken T: The role of the neuropeptide Y (NPY) family in the pathophysiology of inflammatory bowel disease (IBD). Neuropeptides. 55:137–144. 2016. View Article : Google Scholar

4 

El-Salhy M, Danielsson A, Stenling R and Grimelius L: Colonic endocrine cells in inflammatory bowel disease. J Intern Med. 242:413–419. 1997. View Article : Google Scholar

5 

El-Salhy M, Gundersen D, Hatlebakk JG and Hausken T: Chromogranin A cell density as a diagnostic marker for lymphocytic colitis. Dig Dis Sci. 57:3154–3159. 2012. View Article : Google Scholar : PubMed/NCBI

6 

El-Salhy M, Gundersen D, Hatlebakk JG and Hausken T: High densities of serotonin and peptide YY cells in the colon of patients with lymphocytic colitis. World J Gastroenterol. 18:6070–6075. 2012. View Article : Google Scholar : PubMed/NCBI

7 

El-Salhy M, Lomholt-Beck B and Gundersen TD: High chromogranin A cell density in the colon of patients with lymphocytic colitis. Mol Med Rep. 4:603–605. 2011.PubMed/NCBI

8 

Moran GW, Pennock J and McLaughlin JT: Enteroendocrine cells in terminal ileal Crohn's disease. J Crohn's Colitis. 6:871–880. 2012. View Article : Google Scholar

9 

Moran GW, Leslie FC and McLaughlin JT: Crohn's disease affecting the small bowel is associated with reduced appetite and elevated levels of circulating gut peptides. Clin Nutr. 32:404–411. 2013. View Article : Google Scholar

10 

Besterman HS, Mallinson CN, Modigliani R, Christofides ND, Pera A, Ponti V, Sarson DL and Bloom SR: Gut hormones in inflammatory bowel disease. Scand J Gastroenterol. 18:845–852. 1983. View Article : Google Scholar : PubMed/NCBI

11 

El-Salhy M, Mazzawi T, Gundersen D, Hatlebakk JG and Hausken T: The role of peptide YY in gastrointestinal diseases and disorders (Review). Int J Mol Med. 31:275–282. 2013.PubMed/NCBI

12 

Hirotani Y, Mikajiri K, Ikeda K, Myotoku M and Kurokawa N: Changes of the peptide YY levels in the intestinal tissue of rats with experimental colitis following oral administration of mesalazine and prednisolone. Yakugaku Zasshi. 128:1347–1353. 2008. View Article : Google Scholar : PubMed/NCBI

13 

Vona-Davis LC and McFadden DW: NPY family of hormones: Clinical relevance and potential use in gastrointestinal disease. Curr Top Med Chem. 7:1710–1720. 2007. View Article : Google Scholar : PubMed/NCBI

14 

El-Salhy M, Suhr O and Danielsson A: Peptide YY in gastrointestinal disorders. Peptides. 23:397–402. 2002. View Article : Google Scholar : PubMed/NCBI

15 

Tari A, Teshima H, Sumii K, Haruma K, Ohgoshi H, Yoshihara M, Kajiyama G and Miyachi Y: Peptide YY abnormalities in patients with ulcerative colitis. Jpn J Med. 27:49–55. 1988. View Article : Google Scholar : PubMed/NCBI

16 

Sciola V, Massironi S, Conte D, Caprioli F, Ferrero S, Ciafardini C, Peracchi M, Bardella MT and Piodi L: Plasma chromogranin a in patients with inflammatory bowel disease. Inflamm Bowel Dis. 15:867–871. 2009. View Article : Google Scholar

17 

Bishop AE, Pietroletti R, Taat CW, Brummelkamp WH and Polak JM: Increased populations of endocrine cells in Crohn's ileitis. Virchows Arch A Pathol Anat Histopathol. 410:391–396. 1987. View Article : Google Scholar : PubMed/NCBI

18 

Manocha M and Khan WI: Serotonin and GI Disorders: An update on clinical and experimental studies. Clin Transl Gastroenterol. 3:e132012. View Article : Google Scholar : PubMed/NCBI

19 

Stoyanova II and Gulubova MV: Mast cells and inflammatory mediators in chronic ulcerative colitis. Acta Histochem. 104:185–192. 2002. View Article : Google Scholar : PubMed/NCBI

20 

Yamamoto H, Morise K, Kusugami K, Furusawa A, Konagaya T, Nishio Y, Kaneko H, Uchida K, Nagai H, Mitsuma T and Nagura H: Abnormal neuropeptide concentration in rectal mucosa of patients with inflammatory bowel disease. J Gastroenterol. 31:525–532. 1996. View Article : Google Scholar : PubMed/NCBI

21 

Payer J, Huorka M, Duris I, Mikulecky M, Kratochvílová H, Ondrejka P and Lukác L: Plasma somatostatin levels in ulcerative colitis. Hepatogastroenterology. 41:552–553. 1994.PubMed/NCBI

22 

Watanabe T, Kubota Y, Sawada T and Muto T: Distribution and quantification of somatostatin in inflammatory disease. Dis Colon Rectum. 35:488–494. 1992. View Article : Google Scholar : PubMed/NCBI

23 

Koch TR, Carney JA, Morris VA and Go VL: Somatostatin in the idiopathic inflammatory bowel diseases. Dis Colon Rectum. 31:198–203. 1988. View Article : Google Scholar : PubMed/NCBI

24 

Mazzawi T and El-Salhy M: Changes in small intestinal chromogranin A-immunoreactive cell densities in patients with irritable bowel syndrome after receiving dietary guidance. Int J Mol Med. 37:1247–1253. 2016.PubMed/NCBI

25 

El-Salhy M, Umezawa K, Hatlebakk JG and Gilja OH: Abnormal differentiation of stem cells into enteroendocrine cells in rats with DSS-induced colitis. Mol Med Rep. In press.

26 

Elson CO, Sartor RB, Tennyson GS and Riddell RH: Experimental models of inflammatory bowel disease. Gastroenterology. 109:1344–1367. 1995. View Article : Google Scholar : PubMed/NCBI

27 

El-Salhy M and Umezawa K: Treatment with novel AP-1 and NF-κB inhibitors restores the colonic endocrine cells to normal levels in rats with DSS-induced colitis. Int J Mol Med. 37:556–564. 2016.PubMed/NCBI

28 

El-Salhy M, Wendelbo IH, Gundersen D, Hatlebakk JG and Hausken T: Evaluation of the usefulness of colonoscopy with mucosal biopsies in the follow-up of TNBS-induced colitis in rats. Mol Med Rep. 8:446–450. 2013.PubMed/NCBI

29 

Ota E, Takeiri M, Tachibana M, Ishikawa Y, Umezawa K and Nishiyama S: Synthesis and biological evaluation of molecular probes based on the 9-methylstreptimidone derivative DTCM-glutarimide. Bioorg Med Chem Lett. 22:164–167. 2012. View Article : Google Scholar

30 

Takeiri M, Tachibana M, Kaneda A, Ito A, Ishikawa Y, Nishiyama S, Goto R, Yamashita K, Shibasaki S, Hirokata G, et al: Inhibition of macrophage activation and suppression of graft rejection by DTCM-glutarimide, a novel piperidine derived from the antibiotic 9-methylstreptimidone. Inflamm Res. 60:879–888. 2011. View Article : Google Scholar : PubMed/NCBI

31 

Ishikawa Y, Tachibana M, Matsui C, Obata R, Umezawa K and Nishiyama S: Synthesis and biological evaluation on novel analogs of 9-methylstreptimidone, an inhibitor of NF-kappaB. Bioorg Med Chem Lett. 19:1726–1728. 2009. View Article : Google Scholar : PubMed/NCBI

32 

Ueki S, Yamashita K, Aoyagi T, Haga S, Suzuki T, Itoh T, Taniguchi M, Shimamura T, Furukawa H, Ozaki M, et al: Control of allograft rejection by applying a novel nuclear factor-kappaB inhibitor, dehydroxymethylepoxyquinomicin. Transplantation. 82:1720–1727. 2006. View Article : Google Scholar

33 

Matsumoto N, Ariga A, To-e S, Nakamura H, Agata N, Hirano S, Inoue J and Umezawa K: Synthesis of NF-kappaB activation inhibitors derived from epoxyquinomicin C. Bioorg Med Chem Lett. 10:865–869. 2000. View Article : Google Scholar : PubMed/NCBI

34 

Umezawa N, Matsumoto N, Iwama S, Kato N and Higuchi T: Facile synthesis of peptide-porphyrin conjugates: Towards artificial catalase. Bioorg Med Chem. 18:6340–6350. 2010. View Article : Google Scholar : PubMed/NCBI

35 

Hunter MM, Wang A, Hirota CL and McKay DM: Neutralizing anti-IL-10 antibody blocks the protective effect of tapeworm infection in a murine model of chemically induced colitis. J Immunol. 174:7368–7375. 2005. View Article : Google Scholar : PubMed/NCBI

36 

Khan WI and Ghia JE: Gut hormones: Emerging role in immune activation and inflammation. Clin Exp Immunol. 161:19–27. 2010.PubMed/NCBI

37 

Margolis KG and Gershon MD: Neuropeptides and inflammatory bowel disease. Curr Opin Gastroenterol. 25:503–511. 2009. View Article : Google Scholar : PubMed/NCBI

38 

Bampton PA and Dinning PG: High resolution colonic manometry - what have we learnt? - A review of the literature 2012. Curr Gastroenterol Rep. 15:3282013. View Article : Google Scholar

39 

Ameri P and Ferone D: Diffuse endocrine system, neuroendocrine tumors and immunity: What's new? Neuroendocrinology. 95:267–276. 2012. View Article : Google Scholar : PubMed/NCBI

40 

Farzi A, Reichmann F and Holzer P: The homeostatic role of neuropeptide Y in immune function and its impact on mood and behaviour. Acta Physiol (Oxf). 213:603–627. 2015. View Article : Google Scholar

41 

Spångéus A, Forsgren S and el-Salhy M: Does diabetic state affect co-localization of peptide YY and enteroglucagon in colonic endocrine cells? Histol Histopathol. 15:37–41. 2000.PubMed/NCBI

42 

Pyarokhil AH, Ishihara M, Sasaki M and Kitamura N: The developmental plasticity of colocalization pattern of peptide YY and glucagon-like peptide-1 in the endocrine cells of bovine rectum. Biomed Res. 33:35–38. 2012. View Article : Google Scholar : PubMed/NCBI

43 

Mortensen K, Christensen LL, Holst JJ and Orskov C: GLP-1 and GIP are colocalized in a subset of endocrine cells in the small intestine. Regul Pept. 114:189–196. 2003. View Article : Google Scholar : PubMed/NCBI

44 

El-Salhy M, Wilander E and Grimelius L: Immunocytochemical localization of gastric inhibitory peptide (GIP) in the human foetal pancreas. Ups J Med Sci. 87:81–85. 1982. View Article : Google Scholar : PubMed/NCBI

45 

Svendsen B and Holst JJ: Regulation of gut hormone secretion. Studies using isolated perfused intestines. Peptides. 77:47–53. 2016. View Article : Google Scholar

46 

Svendsen B, Pedersen J, Albrechtsen NJ, Hartmann B, Toräng S, Rehfeld JF, Poulsen SS and Holst JJ: An analysis of cosecretion and coexpression of gut hormones from male rat proximal and distal small intestine. Endocrinology. 156:847–857. 2015. View Article : Google Scholar

47 

Egerod KL, Engelstoft MS, Grunddal KV, Nøhr MK, Secher A, Sakata I, Pedersen J, Windeløv JA, Füchtbauer EM, Olsen J, et al: A major lineage of enteroendocrine cells coexpress CCK, secretin, GIP, GLP-1, PYY, and neurotensin but not somatostatin. Endocrinology. 153:5782–5795. 2012. View Article : Google Scholar : PubMed/NCBI

48 

Cardoso WV and Lü J: Regulation of early lung morphogenesis: Questions, facts and controversies. Development. 133:1611–1624. 2006. View Article : Google Scholar : PubMed/NCBI

49 

Darlington GJ: Molecular mechanisms of liver development and differentiation. Curr Opin Cell Biol. 11:678–682. 1999. View Article : Google Scholar : PubMed/NCBI

50 

Fausto N, Campbell JS and Riehle KJ: Liver regeneration. Hepatology. 43(Suppl 1): S45–S53. 2006. View Article : Google Scholar : PubMed/NCBI

51 

Rawlins EL and Hogan BL: Ciliated epithelial cell lifespan in the mouse trachea and lung. Am J Physiol Lung Cell Mol Physiol. 295:L231–L234. 2008. View Article : Google Scholar : PubMed/NCBI

52 

Barker N and Clevers H: Tracking down the stem cells of the intestine: Strategies to identify adult stem cells. Gastroenterology. 133:1755–1760. 2007. View Article : Google Scholar : PubMed/NCBI

53 

Barker N, van de Wetering M and Clevers H: The intestinal stem cell. Genes Dev. 22:1856–1864. 2008. View Article : Google Scholar : PubMed/NCBI

54 

Barker N, van Es JH, Kuipers J, Kujala P, van den Born M, Cozijnsen M, Haegebarth A, Korving J, Begthel H, Peters PJ and Clevers H: Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature. 449:1003–1007. 2007. View Article : Google Scholar : PubMed/NCBI

55 

Barker N, van Oudenaarden A and Clevers H: Identifying the stem cell of the intestinal crypt: Strategies and pitfalls. Cell Stem Cell. 11:452–460. 2012. View Article : Google Scholar : PubMed/NCBI

56 

Cheng C, Chan AO, Hui WM and Lam SK: Coping strategies, illness perception, anxiety and depression of patients with idiopathic constipation: A population-based study. Aliment Pharmacol Ther. 18:319–326. 2003. View Article : Google Scholar : PubMed/NCBI

57 

Rawdon BB and Andrew A: Origin and differentiation of gut endocrine cells. Histol Histopathol. 8:567–580. 1993.PubMed/NCBI

58 

Hoffman J, Kuhnert F, Davis CR and Kuo CJ: Wnts as essential growth factors for the adult small intestine and colon. Cell Cycle. 3:554–557. 2004. View Article : Google Scholar : PubMed/NCBI

59 

Korinek V, Barker N, Moerer P, van Donselaar E, Huls G, Peters PJ and Clevers H: Depletion of epithelial stem-cell compartments in the small intestine of mice lacking Tcf-4. Nat Genet. 19:379–383. 1998. View Article : Google Scholar : PubMed/NCBI

60 

Montgomery RK and Breault DT: Small intestinal stem cell markers. J Anat. 213:52–58. 2008. View Article : Google Scholar : PubMed/NCBI

61 

Potten CS1, Booth C, Tudor GL, Booth D, Brady G, Hurley P, Ashton G, Clarke R, Sakakibara S and Okano H: Identification of a putative intestinal stem cell and early lineage marker; musashi-1. Differentiation. 71:28–41. 2003. View Article : Google Scholar : PubMed/NCBI

62 

Kayahara T, Sawada M, Takaishi S, Fukui H, Seno H, Fukuzawa H, Suzuki K, Hiai H, Kageyama R, Okano H and Chiba T: Candidate markers for stem and early progenitor cells, Musashi-1 and Hes1, are expressed in crypt base columnar cells of mouse small intestine. FEBS Lett. 535:131–135. 2003. View Article : Google Scholar : PubMed/NCBI

63 

He XC, Yin T, Grindley JC, Tian Q, Sato T, Tao WA, Dirisina R, Porter-Westpfahl KS, Hembree M, Johnson T, et al: PTEN-deficient intestinal stem cells initiate intestinal polyposis. Nat Genet. 39:189–198. 2007. View Article : Google Scholar : PubMed/NCBI

64 

Yang Q, Bermingham NA, Finegold MJ and Zoghbi HY: Requirement of Math1 for secretory cell lineage commitment in the mouse intestine. Science. 294:2155–2158. 2001. View Article : Google Scholar : PubMed/NCBI

65 

Jenny M, Uhl C, Roche C, Duluc I, Guillermin V, Guillemot F, Jensen J, Kedinger M and Gradwohl G: Neurogenin3 is differentially required for endocrine cell fate specification in the intestinal and gastric epithelium. EMBO J. 21:6338–6347. 2002. View Article : Google Scholar : PubMed/NCBI

66 

Wang J, Cortina G, Wu SV, Tran R, Cho JH, Tsai MJ, Bailey TJ, Jamrich M, Ament ME, Treem WR, et al: Mutant neurogenin-3 in congenital malabsorptive diarrhea. N Engl J Med. 355:270–280. 2006. View Article : Google Scholar : PubMed/NCBI

67 

Lee CS, Perreault N, Brestelli JE and Kaestner KH: Neurogenin 3 is essential for the proper specification of gastric enteroendocrine cells and the maintenance of gastric epithelial cell identity. Genes Dev. 16:1488–1497. 2002. View Article : Google Scholar : PubMed/NCBI

68 

Naya FJ, Huang HP, Qiu Y, Mutoh H, DeMayo FJ, Leiter AB and Tsai MJ: Diabetes, defective pancreatic morphogenesis, and abnormal enteroendocrine differentiation in BETA2-neuroD-deficient mice. Genes Dev. 11:2323–2334. 1997. View Article : Google Scholar : PubMed/NCBI

69 

Ahlgren U, Jonsson J and Edlund H: The morphogenesis of the pancreatic mesenchyme is uncoupled from that of the pancreatic epithelium in IPF1/PDX1-deficient mice. Development. 122:1409–1416. 1996.PubMed/NCBI

70 

Schonhoff SE, Giel-Moloney M and Leiter AB: Minireview: Development and differentiation of gut endocrine cells. Endocrinology. 145:2639–2644. 2004. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

December 2016
Volume 38 Issue 6

Print ISSN: 1107-3756
Online ISSN:1791-244X

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
El-Salhy, M., Mazzawi, T., Umezawa, K., & Gilja, O.H. (2016). Enteroendocrine cells, stem cells and differentiation progenitors in rats with TNBS-induced colitis. International Journal of Molecular Medicine, 38, 1743-1751. https://doi.org/10.3892/ijmm.2016.2787
MLA
El-Salhy, M., Mazzawi, T., Umezawa, K., Gilja, O. H."Enteroendocrine cells, stem cells and differentiation progenitors in rats with TNBS-induced colitis". International Journal of Molecular Medicine 38.6 (2016): 1743-1751.
Chicago
El-Salhy, M., Mazzawi, T., Umezawa, K., Gilja, O. H."Enteroendocrine cells, stem cells and differentiation progenitors in rats with TNBS-induced colitis". International Journal of Molecular Medicine 38, no. 6 (2016): 1743-1751. https://doi.org/10.3892/ijmm.2016.2787