elafin and Crohn-Disease

More than half of Crohn's disease patients develop intestinal fibrosis-induced intestinal strictures. Elafin is a human protease inhibitor that is down-regulated in the stricturing intestine of Crohn's disease patients. We investigated the efficacy of elafin in reversing intestinal fibrosis and elucidated its mechanism of action.. We developed a new method to mimic a stricturing Crohn's disease environment and induce fibrogenesis using stricturing Crohn's disease patient-derived serum exosomes to condition fresh human intestinal tissues and primary stricturing Crohn's disease patient-derived intestinal fibroblasts. Three mouse models of intestinal fibrosis, including SAMP1/YitFc mice, Salmonella-infected mice, and trinitrobenzene sulfonic acid-treated mice, were also studied. Elafin-Eudragit FS30D formulation and elafin-overexpressing construct and lentivirus were used.. Elafin reversed collagen synthesis in human intestinal tissues and fibroblasts pretreated with Crohn's disease patient-derived serum exosomes. Proteome arrays identified cathepsin S as a novel fibroblast-derived pro-fibrogenic protease. Elafin directly suppressed cathepsin S activity to inhibit protease-activated receptor 2 activity and Zinc finger E-box-binding homeobox 1 expression, leading to reduced collagen expression in intestinal fibroblasts. Elafin overexpression reversed ileal fibrosis in SAMP1/YitFc mice, cecal fibrosis in Salmonella-infected mice, and colonic fibrosis in trinitrobenzene sulfonic acid-treated mice. Cathepsin S, protease-activated receptor 2 agonist, and zinc finger E-box-binding homeobox 1 overexpression abolished the anti-fibrogenic effect of elafin in fibroblasts and all 3 mouse models of intestinal fibrosis. Oral elafin-Eudragit FS30D treatment abolished colonic fibrosis in trinitrobenzene sulfonic acid-treated mice.. Elafin suppresses collagen synthesis in intestinal fibroblasts via cathepsin S-dependent protease-activated receptor 2 inhibition and decreases zinc finger E-box-binding homeobox 1 expression. The reduced collagen synthesis leads to the reversal of intestinal fibrosis. Thus, modified elafin may be a therapeutic approach for intestinal fibrosis.

Topics: Animals; Cathepsins; Collagen; Constriction, Pathologic; Crohn Disease; Elafin; Fibrosis; Humans; Intestinal Obstruction; Intestines; Mice; Peptide Hydrolases; Polymethacrylic Acids; Protease Inhibitors; Proteome; Receptor, PAR-2; Trinitrobenzenesulfonic Acid; Zinc Finger E-box-Binding Homeobox 1

Antimicrobial peptide expression is associated with disease activity in inflammatory bowel disease (IBD) patients. IBD patients have abnormal expression of elafin, a human elastase-specific protease inhibitor and antimicrobial peptide. We determined elafin expression in blood, intestine, and mesenteric fat of IBD and non-IBD patients.. Serum samples from normal and IBD patients were collected from two UCLA cohorts. Surgical resection samples of human colonic and mesenteric fat tissues from IBD and non-IBD (colon cancer) patients were collected from Cedars-Sinai Medical Center.. High serum elafin levels were associated with a significantly elevated risk of intestinal stricture in Crohn's disease (CD) patients. Microsoft Azure Machine learning algorithm using serum elafin levels and clinical data identified stricturing CD patients with high accuracy. Serum elafin levels had weak positive correlations with clinical disease activity (Partial Mayo Score and Harvey Bradshaw Index), but not endoscopic disease activity (Mayo Endoscopic Subscore and Simple Endoscopic Index for CD) in IBD patients. Ulcerative colitis (UC) patients had high serum elafin levels. Colonic elafin mRNA and protein expression were not associated with clinical disease activity and histological injury in IBD patients, but stricturing CD patients had lower colonic elafin expression than non-stricturing CD patients. Mesenteric fat in stricturing CD patients had significantly increased elafin mRNA and protein expression, which may contribute to high circulating elafin levels. Human mesenteric fat adipocytes secrete elafin protein.. High circulating elafin levels are associated with the presence of stricture in CD patients. Serum elafin levels may help identify intestinal strictures in CD patients.

Topics: Abdominal Fat; Adipocytes; Adult; Case-Control Studies; Cell Line; Colitis, Ulcerative; Colon; Colonoscopy; Constriction, Pathologic; Crohn Disease; Elafin; Female; Fibroblasts; Healthy Volunteers; Humans; Intestinal Mucosa; Intestinal Obstruction; Male; Primary Cell Culture; Prospective Studies; Severity of Illness Index

The expression of elafin in inflammatory bowel disease (IBD) is controversial. Here, we detected the expression of elafin in the peripheral blood and colonic mucosa of patient with IBD and then explored its role and value in assessing the activity and severity of IBD.. Sixty-eight patients with IBD were selected as an experimental group. The control group included 38 healthy individuals. The expression of elafin mRNA in peripheral blood leukocytes and in serum was detected by qRT-PCR and enzyme-linked immunosorbent assay, respectively. The inflamed and noninflamed tissues were collected by colonoscopy. The expression of elafin in the intestinal mucosa was determined by immunohistochemistry staining and qRT-PCR. The expression of elafin between groups and among each stage of IBD was compared. The correlations of elafin expression with erythrocyte sedimentation rate and C-reactive protein were determined by Spearman's correlation analysis and with clinical disease activity indices (Best Crohn's Disease Activity Index and modified Mayo scores) by Pearson's correlation analysis.. Elafin mRNA levels decreased significantly in active ulcerative colitis (UC) but increased in remission UC. However, in Crohn's disease (CD), we did not detect the aforementioned significant differences. Although serum IL-8 levels increased, serum elafin concentrations decreased both in UC and in CD, but the differences among stages were not significant. The expression of elafin in the inflamed colonic mucosa in both CD and UC was lower than that in the normal mucosa in controls and lower than that in the noninflamed mucosa in IBD. Moreover, the relative expression of elafin mRNA in peripheral blood leukocytes in UC was negatively correlated with erythrocyte sedimentation rate, C-reactive protein, and modified Mayo scores, and in CD, it was negatively correlated with Best Crohn's Disease Activity Index scores.. Elafin decreased in active patients with IBD and was negatively correlated with disease activity, suggesting that elafin may play a protective role and could be used as an index to evaluate disease activity in IBD.

Topics: Adult; Aged; Blood Sedimentation; C-Reactive Protein; Case-Control Studies; Colitis, Ulcerative; Crohn Disease; Elafin; Female; Humans; Immunohistochemistry; Inflammatory Bowel Diseases; Intestinal Mucosa; Leukocytes; Male; Middle Aged; Real-Time Polymerase Chain Reaction

To quantify the expression of elafin mRNA in peripheral blood in patients with inflammatory bowel disease (IBD) and to explore its value in assessment of the activity and severity of IBD.. From July 1 2015 to August 15 2015, 23 patients with IBD admitted to Peking University First Hospital were selected, including 15 cases with ulcerative colitis (UC) and 8 cases with Crohn's disease (CD). Among those, 5 cases were in remission (UC 3, CD 2), 6 cases were mild active (UC 3, CD 3), 3 cases were moderate active (UC 1, CD 2), and 9 cases were severe active (UC 8, CD 1). A total of 21 healthy individuals were selected as the control group. Peripheral blood samples of IBD patients and healthy controls were collected. The expression of elafin mRNA in peripheral blood leukocytes was detected by fluorescence quantitative real-time PCR. Mann-Whitney test was performed for comparison between the two groups. The correlation between the expression of elafin mRNA in peripheral blood and IBD activity score was analyzed by Pearson correlation analysis after transformation of variables.. The median expression of elafin mRNA in peripheral blood leukocytes in IBD group and control group was 0.005 8 (0.000 2, 0.043 5) and 0.015 3 (0.002 1, 0.175 8), respectively, with no significant difference (P>0.05). However, in the active IBD patients it was lower than that in the controls (0.004 6 (0.000 2, 0.034 8) vs 0.015 3 (0.002 1, 0.175 8), P<0.05) and also lower than that in the remission patients(0.004 6 (0.000 2, 0.034 8) vs 0.023 1 (0.012 6, 0.043 5), P<0.05); in the active UC patients it was lower than that in the controls(0.003 7 (0.000 2, 0.027 0) vs 0.015 3 (0.002 1, 0.175 8), P<0.05). The expression of elafin mRNA in peripheral blood was negatively correlated with modified Mayo score in UC patients (r=-0.513, P<0.05) and with the Crohn's Disease Activity Index (CDAI) of Best score in CD patients (r=-0.889, P<0.05).. The expression of elafin mRNA in peripheral blood in active IBD patients is decreased, which may be correlated with the activity of IBD, and negatively correlated with corresponding disease activity score, suggesting that it may play a protective role in IBD and may be helpful in predicting disease activity.

Topics: Case-Control Studies; Colitis, Ulcerative; Crohn Disease; Elafin; Humans; Inflammatory Bowel Diseases; Leukocytes; Real-Time Polymerase Chain Reaction

Crohn's disease has a genetic background. Onset of the clinical manifestations, however, is suggested to be triggered by environmental factors. Microarray analysis has shown that the expression of transcripts aldolase B, elafin, MST-1, simNIPhom and SLC6A14 are altered in patients with ulcerative colitis. The primary aim of this study was to explore the expressions of these five transcripts in macroscopically inflamed and noninflamed mucosa in patients with Crohn's disease.. Mucosal specimens obtained from colon in consecutive patients with Crohn's disease (n=23) and controls (n=67) undergoing colonoscopy were analyzed using real-time PCR technique.. The expressions of the transcripts aldolase B, elafin, simNIPhom and SLC6A14 were increased, whereas the expression of MST-1 was decreased in noninflamed rectal mucosa in patients with Crohn's disease compared with controls. The expression of aldolase B was increased and the expressions of elafin and simNIPhom were decreased in inflamed colonic mucosa compared with noninflamed rectal mucosa in patients with Crohn's disease. No correlation, between the clinical activity of Crohn's disease (Mayo score or=6) and transcript expression was detected.. The mucosal transcript pattern in Crohn's disease may, based on the known biological function of the transcripts, explain some of the typical features of Crohn's disease and indicate a possible pathophysiological role of microbes. Our results may thereby contribute to the understanding of the pathogenesis and manifestations of Crohn's disease.

Topics: Adult; Aged; Aged, 80 and over; Amino Acid Transport Systems; Amino Acid Transport Systems, Neutral; Case-Control Studies; Colon; Crohn Disease; Elafin; Female; Fructose-Bisphosphate Aldolase; Hepatocyte Growth Factor; Humans; Immunohistochemistry; Intestinal Mucosa; Lymphokines; Male; Middle Aged; Polymerase Chain Reaction; Proto-Oncogene Proteins

Elafin (or skin-derived antileukoprotease) and secretory leukocyte protease inhibitor (SLPI) are serine antiproteases antagonizing human neutrophil elastase (HNE), thereby preventing tissue injury from excessive release of proteolytic enzymes by inflammatory cells. Furthermore, elafin and SLPI are "defensin-like" molecules with broad antimicrobial activity. The balance between proteases and antagonists may critically determine inflammatory processes in Crohn's disease (CD) and ulcerative colitis (UC). Real-time PCR was performed to quantitate colonic, proinflammatory cytokine IL-8, protease (HNE), and antiprotease mRNA (elafin and SLPI) in a total of 340 biopsies from 117 patients (47 CD, 45 UC, 25 controls). Histological inflammation was scored, and HNE, elafin, and SLPI were localized and semiquantified by immunostaining in 51 colonic paraffin sections (23 CD, 11 UC, 17 controls). Proinflammatory IL-8, degree of histological inflammation, and granulocyte content were similar in UC and CD. Elafin stained predominantly in the epithelium and SLPI in mucosal inflammatory cells. HNE mRNA levels and immunostaining were increased equally in both forms of inflammatory bowel disease. Levels of mRNA and immunostaining of the antiproteases elafin and SLPI were enhanced strongly in inflamed versus noninflamed UC. It is surprising that comparing inflamed versus noninflamed CD, this increase was significantly less pronounced for elafin and even lacking for SLPI. Despite comparable degrees of inflammation and protease levels, the induction of both antiproteases was attenuated in CD. This could contribute to the transmural depth of tissue destruction in CD. Elafin and SLPI may be added to the list of defensin-like peptides with diminished induction in CD versus UC.

Topics: Adult; Case-Control Studies; Colon; Crohn Disease; Elafin; Epithelial Cells; Frameshift Mutation; Humans; Inflammation; Intestinal Mucosa; Leukocyte Elastase; Leukocytes; Middle Aged; Nod2 Signaling Adaptor Protein; Secretory Leukocyte Peptidase Inhibitor