| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Departments of Pathology (M.S., T.S., H.S.) and Molecular Biology and Applied Physiology (K.T.), Tohoku University School of Medicine, Sendai, Miyagi 980-8575, Japan; and Division of Gastroenterology and Hepatology, Departments of Internal Medicine (M.S., A.T.), and Pathology, Clinical Service (M.K.), Jikei University School of Medicine, Tokyo 105-8461, Japan
Address all correspondence and requests for reprints to: Masayuki Saruta, M.D., Department of Pathology, Tohoku University School of Medicine, 2-1, Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan. E-mail: m-saruta{at}pg7.so-net.ne.jp.
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
by local mononuclear cells significantly increased in IBD patients (6, 7, 8), and these cytokines are well known to stimulate the hypothalamic-pituitary-adrenal (HPA) axis (9). Continuous chronic inflammation and shallow ulcerations extend proximally from anal margin in patients with UC, and both glucocorticoid treatment (10) and immunosuppressive agents (11) are occasionally effective against patients with UC. Therefore, it is reasonably postulated that some immune abnormalities and/or hypersensitivities may result in colonic mucosal inflammation associated with patients of UC. Urocortin 1 (Ucn1) was recently identified in the rat brain as a neuropeptide of the corticotropin-releasing factor (CRF) family (12). Human (h) Ucn1 has 95% homology in amino acid sequence to rat (r) Ucn1 and 45% to rat/human (r/h) CRF (13). Both immunoreactive Ucn1 (IR-Ucn1) and Ucn1 mRNA have been reported in pituitary glands (14), brain (15), stomach (16), colon (17), heart (18), synovium (19, 20), and lymphocytes (21). Ucn1 has been also found to bind with high affinity to both CRF type 1 (CRF1) and type 2 (CRF2) receptors and has a 6-fold higher affinity for CRF1 than CRF and approximately 40old higher affinity for CRF2 than CRF (12, 13). CRF2 is composed of at least three isoforms, CRF2(a), CRF2(b), and CRF2(c) (22). In humans, CRF2(a) is considered the major CRF2 isoform in skeletal muscle, heart, colon, and brain tissues, and both CRF2(b) and CRF2(c) are considered minor isoforms (17, 23, 24). Therefore, Ucn1 may participate in various physiological CRF receptor-mediated actions via both peripheral and central CRF receptors. Ucn1 also activates the HPA axis and integrates response to stress (12).
In this study, we examined the correlation between in situ production of Ucn1 and intramucosal chronic inflammation in patients with UC. We first studied Ucn1 expression in colonic mucosa with UC using immunohistochemistry. Ucn1-positive cells were further characterized by double immunohistochemistry. Each specimen in the UC group was then graded histopathologically based on the Matts’ classification (25). The number of Ucn1-positive and plasma cells and the ratio of Ucn1-positive cells to plasma cells in each inflammatory grade were compared among these groups. IR-Ucn1 in the colon tissue with UC was further characterized by reverse-phase HPLC coupled with RIA. We subsequently examined the mRNA expression of Ucn1, CRF1, and CRF2 in colonic mucosa with UC using mRNA in situ hybridization to further characterize biological and/or clinical significance of in situ Ucn1 production and actions in patients with UC.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The research protocol for this study was approved by the Ethics Committee of Tohoku University School of Medicine. All colonic mucosal specimens were obtained from endoscopic biopsy. Biopsy specimens of colonic mucosa were retrieved from surgical pathology specimens of 70 patients with UC and 14 nonpathologic specimens at Jikei University Hospital (Tokyo, Japan) and from 14 patients with nonspecific colitis at Tohoku University Hospital (Sendai, Japan). All patients were prepared with oral polyethyleneglycol lavage solutions but not with enema to decrease mechanical stimulation of colon. Clinical characteristics of the patients examined are shown in Table 1
. Forty-one of 70 patients with UC had never received glucocorticoid treatment, and 29 patients with UC were receiving glucocorticoid treatment. The diagnosis of UC was based on history and clinical examination, endoscopic examination, and pathological findings. Nonspecific colitis group consisted of 14 patients with diarrhea and colonoscopic appearance of colitis. These biopsy specimens were diagnosed as nonspecific colitis with hematoxylin and eosin staining. Control group consisted of 14 patients who underwent total colonoscopy with possible diagnosis of polyps. All patients had normal endoscopic findings, and these biopsy specimens were diagnosed as nonsignificant abnormalities with hematoxylin and eosin staining. Biopsy specimens were obtained from several areas of the large intestine (from cecum to rectum) in each subject.
|
Reagents and antisera
hUcn1 (1–40) and r/hCRF (1–41) were commercially obtained from Peptide Institute Inc. (Osaka, Japan). Chemical reagents for mRNA in situ hybridization were all provided from Ventana Medical Systems, Inc. (Tucson, AZ). Other chemicals used in this study were obtained from Wako Chemicals Co. Ltd. (Osaka, Japan). A specific antiserum against Ucn1 was raised in rabbit immunized with a peptide corresponding to rUcn1 (1–40) (Sawady Technology, Tokyo, Japan) (26). This Ucn1 antiserum completely cross-reacted with hUcn1 1–40 but not with other CRF-related peptides (15). VS38c (a monoclonal mouse antihuman plasma cell antibody), CD68 (PG-M1) (a monoclonal mouse antihuman macrophage antibody), and anti-5-hydroxytryptamine (5-HT) antibodies (a monoclonal mouse antihuman serotonin-containing cell antibody) were obtained from Dako A/S (Glostrup, Denmark) (27, 28).
Immunohistochemistry and double-antibody immunostaining
Immunohistochemistry was performed using an Envision system (Dako A/S) and a Histofine immunostaining kit (Nichirei Co. Ltd., Tokyo, Japan) on 3-µm paraffin-embedded sections, as previously reported (14, 17, 18). Optimal dilutions of the antibodies were 1:1000 for Ucn1 and CRF, 1:200 for VS38c, 1:200 for CD68, and 1:20 for 5-HT, respectively.
For absorption test of immunoreactivity, an antibody-antigen mixture containing an optimally diluted antiserum and the peptide (20 µmol/liter, final peptide concentration) was incubated at 4 C overnight. After centrifugation, the resultant supernatants were used as preabsorbed antibodies (14, 17). Cytoplasmic immunoreactivity in lamina propria inflammatory cells obtained with the anti-rUcn11–40 antiserum was prevented by preabsorption with corresponding synthetic peptide, hUcn1 (1–40) but not with r/hCRF (1–41).
For positive identification of immunoreactive cells, double-antibody immunostaining was performed. The following combinations were performed: 1) Ucn1/VS38c; 2) Ucn1/CD68; and 3) Ucn1/5-HT. The first step of immunohistochemistry for Ucn1 was performed with the Envison kit coupled with horseradish peroxidase (Dako A/S) using 3,3'-diaminobenzidine (DAB) solution as a chromogen (brown). After heating these Ucn1-stained tissue sections in a microwave oven for 5 min or placing in trypsin digestion solution for 10 min, the second step immunohistochemistry for VS38c (1:200), CD68 (1:200), or 5-HT (1:20) was carried out with biotin-streptavidin-alkaline phosphatase (Histofine immunostaining kit; Nichirei) using Vector blue (Vector Laboratories, Burlingame, CA) as a chromogen (blue). Positive staining was indicated by brown deposits for the horseradish peroxidase method, blue deposits for the biotin-streptavidin-alkaline phosphatase, and brown-blue deposits for the double-antibody immunostaining. For negative controls, the sections were incubated with normal rabbit IgG instead of the primary antibodies. No specific immunoreactivity was detected in these tissue sections.
Evaluation of positively staining cells
All immunopositive cells were evaluated as positive, regardless of the immunointensity. Both Ucn1- and VS38c-positive cells were counted within the lamina propria areas in each biopsy specimen. The area of lamina propria layer was measured by drawing a boundary around it, using the public domain Image program (version 1.62 f; National Institutes of Health, Bethesda, MD). For each tissue section, cell counts were expressed as cells per square millimeter lamina propria area. The ratio of Ucn1-positive to VS38c-positive cells was subsequently determined. For evaluation, two of the authors (M.S. and T.S.) independently counted Ucn1- and VS38c-positive cells.
To define the quantification of degrees of inflammation, each specimen in the UC groups was stained with hematoxylin and eosin and graded histopathologically based on the widely accepted histopathological classification by Matts (25) (Table 1
). Matts’ grade 5 corresponds to maximum intensity or degree of inflammation, whereas grade 1 implies no inflammation.
Statistical methods
The number of Ucn1-positive and VS38c-positive cells and the ratio for Ucn1-positive cells to plasma cells in each inflammatory grade of UC group were compared among groups, using the Kruskal-Wallis test and Bonferroni/Dunn test. These three categories were compared between glucocorticoid-treated and nonglucocorticoid-treated groups of UC patients using the Mann-Whitney U test.
RIA and HPLC
Tissues were extracted, as reported previously (29). Briefly, the tissue (approximately 750 mg) was boiled in 2 ml of 1 mol/liter acetic acid for 10 min. Eight milliliters of 50% methanol in 0.5 mol/liter acetic acid was added to each sample, and the tissue was homogenized. The homogenate was centrifuged by 15,000 x g for 30 min. The supernatant was separated, dried by air, reconstituted in assay buffer [0.1 mol/liter phosphate buffer (pH 7.5) containing 0.1% (wt/vol) BSA, 0.2% (vol/vol) Triton X-100, and 0.1% (wt/vol) sodium azide] and assayed. IR-Ucn1 in the tissue extract was measured by RIA, as previously reported (15). The assay showed no significant cross-reaction (<0.005%) with r/hCRF, human stresscopin-related peptide, human stresscopin (Peptide Institute), human urocortin III (Phoenix Pharmaceuticals, Inc., Belmont, CA), and other peptides tested.
Chromatographic characterization of tissue extracts was performed by reverse-phase HPLC using a µBondapak C18 column (3.9 mm x 300 mm; Waters, Milford, MA). The tissue extracts of colon samples obtained from UC patients were pooled, reextracted with a Sep-Pak C18 cartridge (Waters), reconstituted in 0.1% (vol/vol) trifluoroacetic acid, and loaded onto the column. Human brain tissue (thalamus) obtained at autopsy was similarly extracted and used as a control. The HPLC was performed with a linear gradient of acetonitrile containing 0.1% (vol/vol) trifluoroacetic acid from 10 to 60% at a flow rate of 1 ml/min·fraction over 50 min. Each fraction (1 ml) was collected, dried by air, reconstituted with assay buffer, and assayed.
mRNA in situ hybridization
mRNA in situ hybridization for Ucn1, CRF1, and CRF2(a) were performed in five UC patients without glucocorticoid treatment using the Discovery (Ventana Medical Systems) (30). Ucn1 cDNA (a 468-bp fragment corresponding to 2236/2954, Gene Bank accession no. AF038633) was amplified from total RNA of human adrenal by RT-PCR, as previously reported (18). CRF1 cDNA (a 475-bp fragment corresponding to 276/750 of the Gene Bank accession no. L23332) and CRF2(a) cDNA (a 233-bp fragment corresponding to 4/236 of the Gene Bank accession no. U34587) were also amplified from total RNA of human brain by RT-PCR. PCR products were ligated to pSPT19 vector (Roche Diagnostics, Mannheim, Germany), yielding the subclone pSPT19-Ucn1, pSPT19-CRF1, and pSPT19-CRF2(a), respectively. The nucleotide sequences were determined using a model 373A autosequencer (Perkin-Elmer, Chiba, Japan) and were confirmed to be identical with the registered sequences. Each plasmid was linealized by the digestion with EcoR1 (for antisense probe) or HindIII (for sense probe). Digoxigenin-labeled RNA probes were generated with digoxigenin-RNA labeling kit (Roche Diagnostics) following the manufacturer’s instruction.
Duplicated slides with the UC colonic mucosa sections were loaded onto the Discovery automated slide-processing system (Ventana Medical Systems). Baking and deparaffinization steps were performed as programmed in the protocol for the RipoMap in situ hybridization regent system (Ventana Medical Systems) on the instrument. In situ hybridization protocols after the deparaffinization step were designed based on the standard protocol described in the manufacturer’s RiboMap application note. The first fixation step was performed using formalin-based RiboPrep reagent (Ventana Medical Systems) for 30 min at 37 C. Reacted sections were subsequently acid treated using hydrochloride-based RiboClear reagent (Ventana Medical Systems) for 10 min at 37 C. The slides were then subjected to protease digestion using protease 2 for 2 min at 37 C. The reacted sections were incubated for hybridization with Ucn1 (3 ng/slide), CRF1 (200 ng/slide), and CRF2(a) (100 ng/slide) antisense RNA probe using RiboHybe hybridization buffer (Ventana Medical Systems) for 6 h at 65 C after a denaturing step for 10 min at 70 C. After three stringency wash steps using 0.1x RiboWash (Ventana Medical Systems; equivalent to 0.2x saline sodium citrate) for 6 min each at 65 C, the second fixation step was performed using RiboPrep reagent for 20 min at 37 C followed by incubation of biotin-labeled antidigoxigenin antibody (Sigma-Aldrich, Inc., St. Louis, MO) for 30 min at 37 C. After streptavidin-alkaline phosphatase conjugate incubation for 16 min at 37 C, the signal was detected automatically using BlueMap NBT/BCIP substrate kit for 6 h at 37 C. Finally, the sections were counterstained with a nuclear fast red for 1 min before covering slipping.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Ucn1 immunoreactivity was markedly detected in cytoplasm of inflammatory cells in the whole lamina propria (Fig. 1A). There were no Ucn1-positive inflammatory cells in colonic solitary lymphatic follicles. Immunoreactivity of VS38c (plasma cell marker) (Fig. 1B) and CD68 (macrophage marker) (Fig. 1C) in serial tissue sections demonstrated that the morphologic features and distribution of Ucn1-positive lamina propria cells were compatible with those of plasma cells but not with those of macrophages. In addition, immunostaining of CRF in serial tissue sections revealed that patterns of localization of Ucn1 immunopositive cells (Fig. 1D) differ from those of CRF immunopositive cells (Fig. 1E). IR-Ucn1 was also detected in mucosal cells near the base of the crypts (Fig. 1F). Immunostaining of 5-HT in serial tissue sections revealed that patterns of localization of Ucn1 immunopositive cells were similar to those of 5-HT-positive serotonin-containing cells (Fig. 1G). There were no significant regional differences of Ucn1 expression in the different portions of the colon examined.
|
). IR-Ucn1 was not detected in macrophages in double-antibody immunostained sections (Fig. 1I). These findings indicate that Ucn1-positive inflammatory cells in the colonic mucosa with UC were mostly lamina propria plasma cells. Serotonin-containing cells near the crypts were especially positive for Ucn1 in double-antibody immunostained sections (Fig. 1J).
The number of Ucn1-positive cells and plasma cells and the ratio of Ucn1-positive cells to plasma cells are summarized in Fig. 2, A–F. In the nonglucocorticoid-treated UC group, the number of Ucn1-positive cells and plasma cells significantly increased in proportion to its inflammation (P < 0.0001; Kruskal-Wallis test) (Fig. 2, A and B). Both Ucn1-positive cells and plasma cells also significantly increased in UC patients with severe grades of inflammation (grade 3 and 4 of the Matts’ classification), compared with the control group (P < 0.0001; Bonferroni/Dunn test) and nonspecific colitis group (P < 0.0001; Bonferroni/Dunn test) (Fig. 2, A and B). The ratio of Ucn1-positive cells to plasma cells were significantly higher in UC patients with the inflammatory grades of 2, 3, and 4 of the Matts’ classification, compared with the control group (P < 0.0001; Bonferroni/Dunn test) (Fig. 2C). There was a positive correlation between the number of Ucn1-positive cells and the overall severity of inflammation and between the number of plasma cells and the overall severity of inflammation. There were no significant differences between nonspecific colitis and the control groups.
|
, D and E). The ratio of Ucn1-positive cells to plasma cells was not higher in UC patients with glucocorticoid treatment than in the control groups (Fig. 2F). The number of Ucn1-positive cells was significantly lower in the patients with noninflammatory grade (grade 1 of Matts’ classification) than the control group (P < 0.0001; Bonferroni/Dunn test). The number of plasma cells was significantly lower in the patients with mild inflammatory grades (grade 1 and 2 of Matts’ classification) than in the control group (P < 0.0001; Bonferroni/Dunn test).
In the glucocorticoid-treated UC group, the number of Ucn1-positive cells was significantly lower in those with each inflammatory grade than the nonglucocorticoid-treated UC group (grade 1; P < 0.05, grades 2–4; P < 0.0001; Mann-Whitney U test) (Fig. 2G). The number of plasma cells was also significantly lower in the patients with severe inflammatory grades than the nonglucocorticoid-treated UC group (grade 3; P < 0.001, grade 4; P < 0.0001; Mann-Whitney’s U test) (Fig. 2H). The ratio of Ucn1-positive cells to plasma cells was significantly lower in the inflammatory grades (grade 2; P < 0.001, grades 3 and 4; P < 0.0001; Mann-Whitney U test) (Fig. 2I).
RIA and HPLC
IR-Ucn1 was detected in four of five colon samples obtained from patients with UC (range 1.3–14.0 pmol/g wet weight), whereas it was detected in only one sample (1.9 pmol/g wet weight) of 16 nonpathologic portions of the large intestine (Fig. 3). Colon samples of UC patients were pooled, and the IR-Ucn1 was further analyzed by reverse-phase HPLC. A small peak eluting in the position of authentic hUcn1 was found in the colon extracts of UC patients (Fig. 4A) as well as in the brain extract (Fig. 4B). A large part of IR-Ucn1 in the UC colon extract and the brain extract was eluted earlier than authentic human Ucn1, forming two or three peaks (from 26th to 35th fraction). Chromatographic profiles of IR-Ucn1 in the tissue extracts were similar between the colon of UC patients and the human brain (Fig. 4).
|
|
In the mucosa from UC patients, accumulation of Ucn1 mRNA hybridization signals was detected in lamina propria inflammatory cells, but not in cells located around the crypts (Fig. 5A). Immunoreactivity of VS38c (Fig. 5C) and CD68 (Fig. 5D) of serial tissue sections demonstrated that the morphologic features and distribution of Ucn1 mRNA-positive lamina propria cells were compatible with those of plasma cells but not with those of macrophages. No significant accumulation of hybridization signals was detected in negative control tissue section using a sense RNA probe for Ucn1 mRNA (Fig. 5B).
|
) and CRF2(a) mRNA (Fig. 5I) hybridization signals was detected in lamina propria inflammatory cells. In serial tissue sections, the morphologic features and distribution of CRF1 and CRF2(a) mRNA-positive lamina propria cells were compatible with those of plasma cells (Fig. 5, G and K) and macrophages (Fig. 5, H and L). No significant accumulations of hybridization signals were detected in negative control tissue sections using sense RNA probes for CRF1 and CRF2(a) mRNA (Fig. 5, F and J).
In serial tissue sections, morphologic features and distribution of Ucn1 mRNA-positive lamina propria plasma cells were similar to those of CRF1 and CRF2(a) mRNA-positive lamina propria cells (Fig. 6, A-C). Ucn1, CRF1, and CRF2(a) mRNAs were partially coexpressed in these lamina propria inflammatory cells.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In addition, both Ucn1-positive cells and plasma cells increased significantly in proportion to inflammatory grades in Matts’ classification in patients with UC without glucocorticoid treatment but not in those with glucocorticoid treatment. These findings demonstrated a positive correlation between local Ucn1 expression and severity of inflammation in the colonic mucosa of patients with UC who had never received glucocorticoid treatment. In our previous study (17), Ucn1-positive cells in the normal colonic mucosa were mainly macrophages. The differences from results of our present study may be due to the presence or the absence of the inflammation, but it awaits further investigations for clarification. IR-Ucn1 was also detected in mucosal cells around the base of the crypts, which correspond to 5-HT-positive enterochromaffin cells.
Plasma cells have been well known to play very important roles in the pathogenesis of UC. Previous reports showed that all types of plasma cells were significantly increased in number in the patients with UC (31), and the dense infiltration of plasma cells in the lower one third of colonic mucosa was independently associated with a shorter time to relapse (32). Plasma cells have been also reported to produce peptides like GH and proinflammatory cytokines such as IL-6 (33, 34). However, this is the first study to demonstrate that plasma cells produced Ucn1 in colonic mucosa of the patients with UC. Previous studies also showed that CRF was expressed in enterochromaffin cells in human normal colonic mucosa (35) and macrophages and epithelial cells of the colonic mucosa in patients with UC (36). This study also demonstrated that Ucn1 expression in the colonic mucosa of patients with UC is different from CRF expression (Fig. 1, D and E).
Consistent with the findings in immunohistochemistry, RIA showed the presence of IR-Ucn1 in four of five colon samples obtained from patients with UC, whereas IR-Ucn1 was not detected in control samples of the large intestine except one sample. These findings indicated that IR-Ucn1 concentrations were elevated in the colon tissue with UC. On the other hand, immunohistochemistry showed the presence of Ucn1-positive cells in nonpathologic colonic specimen. This may be due to a higher sensitivity of the method of immunohistochemistry for Ucn1. Reverse-phase HPLC confirmed the presence of the material eluting in the same position as authentic hUcn1 in the pooled colon extracts of the UC patients. The nature of materials eluting earlier on HPLC remains to be clarified. Such materials eluting in two or three earlier peaks were also found in brain (Fig. 4B) and heart extracts (18) and may represent larger molecular-weight forms of Ucn1, probably the precursor of Ucn1, precursor fragments, or their modified form(s). Ucn1 may be stored mainly in larger molecular forms in these tissues, including colonic mucosa with UC. On the other hand, we could not completely exclude the possibility that the Ucn1 antiserum used in the present study had a higher affinity to such larger molecular forms of Ucn1 described above.
mRNA in situ hybridization demonstrated that two subtypes of CRF receptors, CRF1 and CRF2(a), were coexpressed in lamina propria inflammatory cells in patients with UC. In serial tissue sections, these inflammatory cells corresponded to plasma cells and macrophages. In addition, these mRNA hybridization signals were detected in lamina propria macrophages but not in superficial macrophages. Morphologic features and distribution of Ucn1 mRNA-positive lamina propria plasma cells were identical with CRF1 and CRF2(a) mRNAs-positive lamina propria cells. On the other hand, we could not exclude the possibility that CRF2(b) mRNA was expressed in CRF2(a) mRNA-positive cells because the probe for CRF2(a) (4/236, Gene Bank accession no. U34587) used in the present study shares the nucleotide sequences of CRF2(b) (103/236, Gene Bank accession no. U34587). These results all suggest that in the colonic mucosa with chronic active inflammation, plasma cells actively produce Ucn1, which may act on both plasma cells and macrophages as autocrine/paracrine mediators via CRF receptors.
Both plasma cells and macrophages produce proinflammatory cytokines, such as IL-1ß, IL-6, and TNF. Results of recent studies demonstrated that Ucn1 enhanced production of IL-1ß and IL-6 in cultured peripheral blood mononuclear cells (19), and IL-1ß, IL-6, and TNF play important roles to induce its inflammatory activity in patients with IBD (6, 7, 8). IL-1ß, IL-6, and TNF are all multifunctional proinflammatory cytokines. IL-1ß and IL-6 both act to induce acute phase protein synthesis in hepatocytes (37, 38), and IL-6 acts to induce terminal differentiation of activated B cells into antibody producing cells, i.e. plasma cells (38). In addition, these proinflammatory cytokines stimulate secretion of CRF from the hypothalamus and ACTH from the pituitary (9). Ucn1 secreted by plasma cells may stimulate the production and/or secretion of some proinflammatory cytokines in the autocrine or paracrine fashion. In addition to enhancing local inflammatory actions in colonic mucosa, these cytokines may also activate the HPA axis and increase the secretion of glucocorticoid, which in turn suppresses the inflammation of UC. In addition, glucocorticoid treatment suppressed the proliferation of plasma cells and the ratio of Ucn1-positive cells to plasma cells, resulting in decreased Ucn1 expression, which is consistent with the hypothesis above.
Recent studies suggest that Ucn1 plays important roles in regulation of local inflammation in many different tissues. Ucn1 has been reported to work as proinflammatory agent in rheumatoid arthritis (19, 20), whereas another report demonstrated that Ucn1 exerts its effects as antiinflammatory agent in Helicobacter pylori-related gastritis (16). The CRF2-mediated actions of Ucn1 are mostly antiinflammatory, i.e. inhibition of heat-induced edema (39) and protection of cardiac myocytes from cell death induced by hypoxia (40). Recently Luckey et al. (41) demonstrated CRF1 might mediate postoperative gastric ileus. Ucn1 in the colonic mucosa with UC may therefore exert proinflammatory effects through the release of proinflammatory cytokines possibly via CRF1, whereas Ucn1 may work against the inflammation via CRF2 in the colon and the HPA axis activation together with some cytokines.
In conclusion, we demonstrated that Ucn1 is locally synthesized in both lamina propria plasma cells and enterochromaffin cells in patients with UC, and the number of Ucn1-positive cells and plasma cells and the ratio of Ucn1-positive cells to plasma cells all increased corresponding to the inflammatory activity, but glucocorticoid treatment suppressed these cells and the ratio above. These findings indicate that Ucn1 may act as an immune-inflammatory mediator in colonic mucosa with UC.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
Abbreviations: CRF, Corticotropin-releasing factor; CRF1, CRF type 1; CRF2, CRF type 2; DAB, 3,3'-diaminobenzidine; h, human; HPA, hypothalamic-pituitary-adrenal; 5-HT, 5-hydroxytryptamine; IBD, inflammatory bowel disease; IR-Ucn1, immunoreactive Ucn1; r, rat; UC, ulcerative colitis; Ucn1, urocortin 1.
Received February 5, 2004.
Accepted August 6, 2004.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
messenger RNA in the intestinal mucosa of inflammatory bowel disease, particularly in patients with disease in the inactive phase. J Gastroenterol 37:345–353[CrossRef][Medline]
receptor. Mol Endocrinol 12:1077–1085This article has been cited by other articles:
 |
|
 |
|
  C Wallon, P-C Yang, A V Keita, A-C Ericson, D M McKay, P M Sherman, M H Perdue, and J D Soderholm Corticotropin-releasing hormone (CRH) regulates macromolecular permeability via mast cells in normal human colonic biopsies in vitro Gut, January 1, 2008; 57(1): 50 - 58. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Kimura, T. Amano, H. Uehara, H. Ariga, T. Ishida, A. Torii, H. Tajiri, K. Matsueda, and S. Yamato Urocortin I is present in the enteric nervous system and exerts an excitatory effect via cholinergic and serotonergic pathways in the rat colon Am J Physiol Gastrointest Liver Physiol, October 1, 2007; 293(4): G903 - G910. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C Moss, P. Anton, T. Savidge, P. Newman, A. S Cheifetz, J. Gay, S. Paraschos, M. W. Winter, M. P Moyer, K. Karalis, et al. Urocortin II mediates pro-inflammatory effects in human colonocytes via corticotropin-releasing hormone receptor 2{alpha} Gut, September 1, 2007; 56(9): 1210 - 1217. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E Zoumakis, D K Grammatopoulos, and G P Chrousos Corticotropin-releasing hormone receptor antagonists Eur. J. Endocrinol., November 1, 2006; 155(suppl_1): S85 - S91. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Kokkotou, D. Torres, A. C. Moss, M. O'Brien, D. E. Grigoriadis, K. Karalis, and C. Pothoulakis Corticotropin-releasing hormone receptor 2-deficient mice have reduced intestinal inflammatory responses. J. Immunol., September 1, 2006; 177(5): 3355 - 3361. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E Gonzalez-Rey, A Fernandez-Martin, A Chorny, and M Delgado Therapeutic effect of urocortin and adrenomedullin in a murine model of Crohn's disease Gut, June 1, 2006; 55(6): 824 - 832. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Gonzalez-Rey, A. Chorny, N. Varela, G. Robledo, and M. Delgado Urocortin and Adrenomedullin Prevent Lethal Endotoxemia by Down-Regulating the Inflammatory Response Am. J. Pathol., June 1, 2006; 168(6): 1921 - 1930. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Wu, Y. Xu, H. Zhou, J. Tao, and S. Li Expression of urocortin in rat lung and its effect on pulmonary vascular permeability. J. Endocrinol., April 1, 2006; 189(1): 167 - 178. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Fukuda, K. Takahashi, T. Suzuki, M. Saruta, M. Watanabe, T. Nakata, and H. Sasano Urocortin 1, Urocortin 3/Stresscopin, and Corticotropin-Releasing Factor Receptors in Human Adrenal and Its Disorders J. Clin. Endocrinol. Metab., August 1, 2005; 90(8): 4671 - 4678. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS |
