interleukin-8 and Colitis--Ulcerative

Ulcerative colitis (UC) is a chronic disease affecting the large intestine. Cytokines, as inflammatory mediators, can enable pathological injury of the intestinal mucosa and play an important role in UC's pathogenesis. Traditional Chinese medicine (TCM) offers a wealth of theory and experience in UC's treatment.. The literature review and meta-analysis intended to examine TCM's effects in the treatment of UC patients who have the dampness-heat syndrome on the serum cytokines known to be related to UC's pathogenesis.. The research team conducted a comprehensive literature search for randomized controlled trials (RCTs) in seven databases. The search covered all publicly published documents from the establishment of a database until August 31, 2021. The team also performed a meta-analysis of the RCTs' results to compare the levels of cytokines in the intervention and control groups.. The study took place at Yueyang Hospital of Integrated Traditional Chinese and Western Medicine of Shanghai University of Traditional Chinese Medicine in Shanghai, China.. For the meta-analysis, the research team created two intervention groups, the oral TCM only group and the TCM+ Western Medicine (WM) group and a control group, the WM group. The team determined which RCT's measured a particular cytokine and which groups those RCTs compared, the team examined the differences between the groups postintervention.. The primary outcome measures were the RCTs' levels of 13 serum cytokines-interleukin 6 (IL-6), IL-8, tumor necrosis factor alpha (TNF-α), IL-17, IL-23, interferon-gamma (IFN-γ), IL-21, IL-1, IL-1β, IL-2, IL-4, IL-10, and IL-13. The team used the random effects model to combine the results for the serum markers as standardized mean differences (SMDs) and compared the two intervention groups to the control group.. The research team identified 22 studies that included 1957 participants. The team found that six proinflammatory cytokines were significantly lower in the combined TCM only and TCM+WM intervention groups than in the WM control group: (1) IL-6-SMD -2.60, 95%CI -3.37 to -1.83, P < .00001; (2) IL-8-SMD -2.49, 95%CI -3.34 to -1.64, P < .00001; (3) TNF-α-SMD -1.70, 95%CI -2.07 to -1.33, P < .00001; (4) IL-17 (TCM+WM group only)-SMD-2.99, 95%CI -4.66 to -1.31, P = .0005; (5) IL-23 (TCM+WM group only)-SMD -2.43, 95% CI -2.78 to -2.08, P < .00001; and (6) IFN-γ-SMD -1.47, 95% CI -1.81 to -1.12, P < .00001. The team found that two anti-inflammatory cytokines were significantly higher in the intervention group than in the control group: (1) IL-4-SMD 1.45, 95% CI 0.92-1.99, P < .00001, and (2) IL-10-SMD 1.33, 95% CI 0.97-1.69, P < .00001. For the results that the team couldn't combine, the levels of the proinflammatory cytokines IL-1, IL-1β, IL-2, and IL-21 were significantly lower in the combined intervention groups than in the control group (P < .05), and the level of the anti-inflammatory cytokine IL-13 in the intervention group was significantly higher than that in the control group (P < .05). The comprehensive analysis showed that oral TCM or a combination of TCM and WM could more significantly reduce the levels of the proinflammatory cytokines IL-6, IL-8, TNF-α, IL-17, IL-23, IFN-γ, IL-21, IL-1, IL-1β and IL-2 and increase the levels of the anti-inflammatory cytokines IL-4, IL-10 and IL-13.. Oral TCM or TCM+WM can reduce the proinflammatory response and increase the anti-inflammatory response of UC patients by regulating serum cytokines and can obtain a better clinical effect than WM only. These benefits can alleviate intestinal inflammation in patients and have a positive effect on clinical efficacy. In the future, more high-quality, large-sample, and long-term follow-up randomized controlled trial are necessary to support research analysis.

Topics: Anti-Inflammatory Agents; China; Colitis, Ulcerative; Cytokines; Hot Temperature; Humans; Interleukin-1; Interleukin-10; Interleukin-13; Interleukin-17; Interleukin-2; Interleukin-23; Interleukin-4; Interleukin-6; Interleukin-8; Medicine, Chinese Traditional; Syndrome; Tumor Necrosis Factor-alpha

To systematically assess effectiveness and safety of Bifidobacterium quadruple viable bacteria combined with mesalamine against ulcerative colitis (UC) in the Asian population.. An electronic search was conducted in PubMed, Embase, Cochrane Library, CNKI, VIP, and Wanfang databases for a random collection of controlled trials of Bifidobacterium quadruple viable bacteria combined with mesalamine against UC. Following data screening and extraction, a Cochrane risk assessment tool was adopted to evaluate the quality of the included studies, and RevMan 5.3 and Stata/SE 15.1 software were used for meta-analysis.. Nineteen articles which enrolled 1,707 subjects were included ultimately in this study. The experimental group performed better than the control group in improving the Mayo score (MD = -1.94, 95% CI = (-2.69, -1.19),. In conclusion, the current meta-analysis shows that Bifidobacterium quadruple viable bacterium combined with mesalamine has a satisfactory effect in the treatment of UC in China, and its safety is better than that of mesalamine or Bifidobacterium quadruple viable bacteria alone. However, randomized controlled trials with standardized designs and large sample sizes are still needed for further validation.

Topics: Bifidobacterium; C-Reactive Protein; Colitis, Ulcerative; Humans; Interleukin-4; Interleukin-8; Mesalamine; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Ulcerative colitis (UC) is a major type of inflammatory bowel disease (IBD), which is characterized by diffuse inflammation of the mucosa of the colon and rectum. Abdominal pain, diarrhea, and hematochezia are UC's main clinical manifestations. Pathogenesis of UC has not yet been clearly elucidated, but it is considered to result from dysregulated expressions of molecules engaged in proinflammatory and anti-inflammatory processes. CXCL8 is one of the most important proinflammatory factors which play a vital role in many inflammatory diseases including UC. The CXCL8-CXCR1/2 axis participates in the pathogenesis of UC through multiple signaling pathways, including PI3k/Akt, MAPKs and NF-κB signaling pathways. Meanwhile, more and more studies in recent years have shown that UC patients have specific non-coding RNA (ncRNA) expression profiles, which may be involved in the occurrence and development of inflammation. In this article, we analyzed the CXCL8-CXCR1/2 axis related signaling pathways and ncRNAs in UC, as well as recent advances in our understanding of the CXCL8-CXCR1/2 axis inhibition as a therapeutic strategy against UC.

Topics: Animals; Colitis, Ulcerative; Humans; Interleukin-8; Intestinal Mucosa; RNA, Untranslated; Signal Transduction

Ulcerative colitis (UC) is associated with chronic relapsing inflammation of the colon. Cytoapheresis is a therapeutic strategy of extracorporeal immunomodulation that has been used in several immunological disorders. In Japan, several open trials of cytoapheresis for UC patients accumulated encouraging results in bringing steroid-resistant patients into remission. In this paper, three types of cytoapheresis [leukocytapheresis (LCAP), granulocytapheresis (GCAP), and centrifugal leukocyte apheresis] are reviewed.

Topics: CD4-Positive T-Lymphocytes; Colitis, Ulcerative; Cytapheresis; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Leukocyte Count; Monocytes; Platelet Count

Inflammatory bowel diseases remain a significant chronic disease affecting children and adolescents. Although corticosteroids remain the standard form of therapy for many patients, an era of biological agents for therapy in inflammatory bowel diseases is upcoming with human trials using these agents now forthcoming. In addition, studies on maintenance of remission are beginning to address the selection of those patients most likely to benefit from aminosalicylate therapies, the risks of relapse from using cyclooxygenase inhibitors, the lack of benefit from lipoxygenase inhibitors, and possible future methodologies to examine the effectiveness of the immuno-suppressive agent 6-mercaptopurine. Further development of the hypothesis that new-onset disease may be different than longstanding disease can be appreciated with reports of cytokine analysis in new-onset inflammation and the high risk of progression of disease in new-onset ulcerative proctitis in children. With more reports on the role of intestinal epithelial cells in intestinal diseases, it is now clear that this cell layer is not a passive bystander with regard to the interactions between the luminal contents and immune system components found within the lamina propria; new studies suggest that novel therapeutic strategies may be possible. This review summarizes recently reported aspects of Crohn's disease and ulcerative colitis, with an emphasis on issues-pertinent for younger patients.

Topics: Anti-Inflammatory Agents; Budesonide; Chemokine CXCL5; Chemokines, CXC; Chemotaxis, Leukocyte; Child; Colitis, Ulcerative; Crohn Disease; Humans; Interleukin-10; Interleukin-8; Nutritional Status

Topics: Colitis, Ulcerative; Crohn Disease; Cytokines; Humans; Interferon-gamma; Interleukin-1; Interleukin-2; Interleukin-6; Interleukin-8; Intestinal Mucosa; Sulfasalazine; Tumor Necrosis Factor-alpha

Present investigation is conducted to investigate the clinical efficacy of mesalazine in combination with the Bifid Triple Viable Capsules on the ulcerative colitis (UC) and the resultant effect on the inflammatory factors (TNF-α, IL-8 and IL-10) of UC patients. A total of 120 UC patients who were admitted to this hospital for treatment between May 2014 and February 2018 were enrolled in this study and divided randomly into the research group and control group, with 60 patients in each group. For patients in the two groups, they underwent medication via mesalazine, while those in the research group additionally received the medication by Bifid Triple Viable Capsules. Following treatment, we evaluated the clinical efficacy, as well as the disease activity index (DAI) of UC, score of clinical symptoms, changes in the inflammatory factors (TNF-α, IL-8 and IL-10) and the adverse reactions to drugs before and after treatment. The total effectiveness rate in the research group was 90.0%, significantly higher than 72.5% in the control group, and the difference had statistical significance (P < 0.05). Before treatment, we assessed the UCDAI and clinical symptoms, and found that there were no statistically significant differences in these indicators between two groups (P>0.05); however, after treatment, both of UCDAI and clinical symptoms scores were decreased evidently in two groups (P<0.05), while the decreases in the research group were more significant (P < 0.05). In addition, following treatment, the levels of TNF-α and IL-8 were all decreased in two groups, with an acute increase in IL-10 (all P<0.01), and the alterations in these indicators in the research group were much more significant than those in the control group (all P <0.05). For adverse reactions, the incidence rate in the research group was 6.67%, significantly lower than 33.33% in the control group (P <0.05). Mesalazine in combination with Bifid Triple Viable Capsules shows a magnificent protective effect on the mucosa of UC patients, and curb the UC-related inflammatory reactions effectively. Thus, it is a safe and reliable method that is worthy of being promoted in clinical practice.

Topics: Adult; Bifidobacterium; Colitis, Ulcerative; Drug Therapy, Combination; Enterococcus; Female; Humans; Interleukin-10; Interleukin-8; Lactobacillus acidophilus; Male; Mesalamine; Probiotics; Severity of Illness Index; Treatment Outcome; Tumor Necrosis Factor-alpha

Treatment of active ulcerative colitis is associated with incomplete efficacy, adverse events, and loss of response. Toll-like receptor-9 mediates innate and adaptive immune response toward intestinal microorganisms. The oral synthetic oligonucleotide toll-like receptor-9 modulator has demonstrated anti-inflammatory properties in colitis murine models and a satisfactory safety profile in humans.. To evaluate the efficacy and safety of BL-7040 (a Toll-like receptor-9 modulator) in patients with moderately active ulcerative colitis.. Moderately active ulcerative colitis patients were included in this multicenter, open-label phase IIa trial. Concomitant mesalamine and steroids at a stable dose were allowed. Clinical outcome was evaluated using the Mayo score, histology, and mucosal cytokine levels. Side effects were registered.. Sixteen out of 22 patients completed a 5-week treatment course and 2-week follow-up. Six patients discontinued the study, three of them due to adverse events. Clinical remission was observed in two patients (12.5 %), and clinical response as well as mucosal healing were achieved in half (50 %) of the patients, while all others remained stable. Furthermore, mucosal neutrophil (p = 0.002) and mucosal interleukin-6 levels (p = 0.046) were significantly reduced in responders compared to non-responders. Toll-like receptor-9 was well tolerated with only one unrelated to study drug serious adverse event (hemoglobin decrease) and 29 mild-to-moderate adverse events.. Oral administration of the Toll-like receptor-9 agonist BL-7040 appeared efficacious, safe and well tolerated in patients with moderately active ulcerative colitis.

Topics: Administration, Oral; Adult; Colitis, Ulcerative; Colon; Female; Humans; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Neutrophils; Oligodeoxyribonucleotides; Toll-Like Receptor 9; Treatment Outcome; Tumor Necrosis Factor-alpha

The efficacy of germinated barley foodstuff (GBF) on tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and -8 (IL-8) in patients with ulcerative colitis (UC) has not yet been examined. The aim of the present study was to determine the effect of administration of GBF on serum TNF-α, IL-6 and -8 levels in UC patients in remission.. Forty-one patients with UC were divided into two groups, namely control and GBF group. Twenty-one patients in the control group received standard treatment while 20 patients in the GBF group received 30 g of GBF daily by oral administration during two months of the study along with standard drug therapy.. Levels of TNF-α, IL-6 and -8 all decreased in the GBF group compared with baseline during the two-month study, while in the control group all values rose. For IL-6 and -8 this effect was significant, P = 0.034 and 0.013, respectively.. The results of the present study showed that the consumption of GBF may reduce the level of serum TNF-α, IL-6 and -8 in patients with UC. This investigation was designed as a pilot study and the results may provide a basis for more future clinical trials.

Topics: Administration, Oral; Adult; Colitis, Ulcerative; Dietary Supplements; Female; Germination; Hordeum; Humans; Interleukin-6; Interleukin-8; Male; Prebiotics; Recurrence; Tumor Necrosis Factor-alpha

Our previous studies have demonstrated the efficacy of partition-herb moxibustion on ulcerative colitis (UC) rats. However, the mechanism of actions of partition-herb moxibustion is still unclear. Most Chinese acupuncturists believe that partition-herb improves the effect of moxibustion therapy. However, this argument lacks confirming experimental and clinical evidence. So, whether partition-herb does play a role in the mechanism of actions of partition-herb moxibustion was reported in this paper. A total of 123 patients were randomly divided into the partition-herb moxibustion group and the control group (partition-bran moxibustion group). Fourteen patients dropped out of the study. Finally, 109 patients finished the intervention. Sixty-one patients with UC received partition-herb moxibustion and 48 patients with UC received partition-bran moxibustion for 72 treatments, one treatment per day, every 12 treatments with an interval of 3 days. The expressions of interleukin-8 (IL-8) and intercellular adhesion molecule-1 (ICAM-1) in the colonic mucosa tissue in mild patients with UC were assayed by immunohistochemistry and RT-PCR. Histology of the colon also was observed at entry and after 72 treatments. Clinical therapeutic effect in partition-herb moxibustion was significantly better than those in partition-bran moxibustion (P < 0.05). Protein and mRNA expression of IL-8, ICAM-1 in the patients was inhibited by both partition-herb moxibustion and partition-bran moxibustion, of those the inhibition in the partition-herb moxibustion group was more obvious (P < 0.01). Partition-herb moxibustion was shown to significantly improve the histology of the colon, and partition-herb was shown to improve the therapeutic effect of moxibustion on treatment of UC patients.

Topics: Adolescent; Adult; Aged; Colitis, Ulcerative; Colon; Down-Regulation; Female; Humans; Immunohistochemistry; Intercellular Adhesion Molecule-1; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Moxibustion; Proteins; RNA, Messenger; Sigmoidoscopy

To evaluate the effectiveness and safety of oral N-acetyl-L-cysteine (NAC) co-administration with mesalamine in ulcerative colitis (UC) patients.. Thirty seven patients with mild to moderate UC were randomized to receive a four-wk course of oral mesalamine (2.4 g/d) plus N-acetyl-L-cysteine (0.8 g/d) (group A) or mesalamine plus placebo (group B). Patients were monitored using the Modified Truelove-Witts Severity Index (MTWSI). The primary endpoint was clinical remission (MTWSI < or = 2) at 4 wk. Secondary endpoints were clinical response (defined as a reduction from baseline in the MTWSI of > or = 2 points) and drug safety. The serum TNF-alpha, interleukin-6, interleukin-8 and MCP-1 were evaluated at baseline and at 4 wk of treatment.. Analysis per-protocol criteria showed clinical remission rates of 63% and 50% after 4 wk treatment with mesalamine plus N-acetyl-L-cysteine (group A) and mesalamine plus placebo (group B) respectively (OR = 1.71; 95% CI: 0.46 to 6.36; P = 0.19; NNT = 7.7). Analysis of variance (ANOVA) of data indicated a significant reduction of MTWSI in group A (P = 0.046) with respect to basal condition without significant changes in the group B (P = 0.735) during treatment. Clinical responses were 66% (group A) vs 44% (group B) after 4 wk of treatment (OR = 2.5; 95% CI: 0.64 to 9.65; P = 0.11; NNT = 4.5). Clinical improvement in group A correlated with a decrease of IL-8 and MCP-1. Rates of adverse events did not differ significantly between both groups.. In group A (oral NAC combined with mesalamine) contrarily to group B (mesalamine alone), the clinical improvement correlates with a decrease of chemokines such as MCP-1 and IL-8. NAC addition not produced any side effects.

Topics: Acetylcysteine; Adult; Aged; Anti-Inflammatory Agents, Non-Steroidal; Chemokine CCL2; Colitis, Ulcerative; Drug Therapy, Combination; Female; Free Radical Scavengers; Glutathione; Humans; Interleukin-6; Interleukin-8; Male; Mesalamine; Middle Aged; Pilot Projects; Severity of Illness Index; Treatment Outcome; Tumor Necrosis Factor-alpha

Adacolumn selective granulocyte and monocyte apheresis (GMA) depletes activated leukocytes in patients with ulcerative colitis (UC). However, this per se cannot fully explain the efficacy of GMA. We have investigated the effects of GMA on the expression of toll-like receptors (TLRs) and plasma interleukin-8 (IL-8). Twenty-two patients with clinical activity index (CAI) of 5-17, 15 with total colitis and 7 with left-sided colitis, were included. Each patient could receive up to 10 GMA sessions, at 1 or 2 sessions per week. GMA was added to the patients' ongoing medication following a relapse or worsening UC, but no additional medication was given. Further, at entry and pre-GMA, blood samples were taken for full blood cell count, expression of TLRs on leukocytes, and plasma IL-8. Seventy-five percent of patients achieved remission after the 10th session (CAI, < or =4; P < 0.005) and there was a marked fall in C-reactive protein (P < 0.01), plasma IL-8 (P < 0.001), and granulocytes (P < 0.05) but an increase in lymphocytes (P < 0.05). The expression of TLR2 on granulocytes was down-modulated (P < 0.05) together with suppression of inflammatory cytokines produced by peripheral blood leukocytes. In conclusion, GMA appears to be an effective adjunct therapy to induce remission in the majority of patients, who are then spared from excess drug therapy. The procedure is associated with sustained immunomodulation. Control studies should strengthen these findings.

Topics: Adolescent; Adult; Colitis, Ulcerative; Cytokines; Female; Granulocytes; Humans; Interleukin-8; Leukapheresis; Leukocytes; Lymphocyte Count; Male; Middle Aged; Toll-Like Receptor 2

Ulcerative colitis is characterized by relapsing mucosal inflammation where the lesions include tissue-damaging granulocytes. In addition, T cells and natural killer (NK) cells play important pathophysiologic roles. Chemokines are a large family of peptides that play key roles in the regulation of inflammation. The CXC-chemokines, growth-related oncogene (GRO)-alpha/CXCL1 and interleukin (IL)-8/CXCL8, both recruit neutrophils and possess mitogenic properties, whereas the interferon-dependent CXC-chemokines monokine induced by gamma-interferon (MIG)/CXCL9, interferon-gamma inducible protein of 10 kD/CXCL10, and IFN-inducible T cell alpha chemoattractant/CXCL11 recruit and activate T cells and NK cells.. The expression of CXC-chemokines was studied in eight controls and in 11 patients suffering from ulcerative colitis in the distal part of the colon, before and during topical treatment with corticosteroids. Perfusates (obtained before, after 7 days, and after 28 days of treatment) and pinch biopsies (obtained before and after 28 days of treatment) were collected by colonoscopy. The rectal release of GRO-alpha and MIG was determined by enzyme-linked immunosorbent assay (ELISA), and tissue expression of the chemokines was detected in colonic tissue by immunohistochemistry.. In perfusates, high levels of GRO-alpha, IL-8, and MIG were detected compared with controls (p=0.02, 0.005, and p=0.03, respectively). During treatment with corticosteroids, both GRO-alpha and MIG decreased. In clinical nonresponders, characterized by sustained inflammation, the levels of GRO-alpha and MIG remained elevated. Both epithelial cells and granulocytes, present in the submucosa, expressed GRO-alpha and MIG as detected by immunohistochemistry.. CXC-chemokines are likely to be important in the pathophysiology of ulcerative colitis and may become targets for novel treatment strategies. In addition, GRO-alpha may serve as a marker of disease activity.

Topics: Administration, Topical; Adrenal Cortex Hormones; Adult; Aged; Anti-Inflammatory Agents; Biomarkers; Chemokine CXCL1; Chemokine CXCL10; Chemokine CXCL9; Colitis, Ulcerative; Colon; Colonoscopy; Down-Regulation; Enema; Enzyme-Linked Immunosorbent Assay; Female; Gastrointestinal Agents; Humans; Immunohistochemistry; Interleukin-8; Male; Middle Aged; Perfusion; Prednisolone; Severity of Illness Index; Time Factors; Treatment Outcome

Enteric microflora of ulcerative colitis patients becomes aberrant. The abnormal interaction between microflora and intestinal mucosal immune system leads the mucosal inflammation. Probiotic administration may recover the commensal microflora and normalise the host-microbial interaction. In this experiment, we cocultured colonic biopsies from active ulcerative colitis patients with bifidobacterium to investigate the modulation effect of probiotics on inflamed colonic tissues and its possible mechanism. Colonic biopsies from active ulcerative colitis were cocultured for 24 h with Bifidobacterium longum. Tumour necrosis factor (TNF)-alpha and interleukin (IL)-8 in supernatants were measured using enzyme-linked immunosorbent assays, the biopsies were fixed using paraffin and the expression of NF-kappaB P65 of tissues was studied using immunohistochemical staining. The concentrations of TNF-alpha and IL-8 in supernatants of tissues cocultured with probiotics were lower than those cultured alone. The number of lamina propria mononuclear cells (LPMC) with nuclear factor-kappa B (NF-kappaB) P65 positive in cocultured tissues was also decreased. When cocultured with inflamed tissues of active ulcerative colitis, probiotics could inhibit NF-kappaB activation in LPMC and down-regulate inflammatory cytokine secretion from inflamed tissues of active ulcerative colitis.

Topics: Adolescent; Adult; Colitis, Ulcerative; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Intestinal Mucosa; L-Lactate Dehydrogenase; Male; Middle Aged; NF-kappa B; Probiotics; Tumor Necrosis Factor-alpha

Our goal was to determine the effect of transdermal nicotine on cytokine and mucin gene transcription in ulcerative colitis (UC). Sixty-four nonsmoking patients with active UC were randomly assigned to transdermal nicotine (maximum dose 22 mg/day) or placebo for 4 weeks. Clinical assessment and colonic mucosal biopsies were obtained at entry and after 4 weeks. Inflammatory and immunoregulatory cytokines were assessed by qualitative reverse transcriptase-polymerase chain reaction (RT-PCR). Based on this initial screen. IL-8 mRNA levels were measured by RT-competitive PCR. MUC1, MUC2, MUC3, MUC4, MUC5AC, MUC5B, and MUC6 mRNA concentrations were measured by quantitative dot blot analysis. Cytokine mRNA expression, except for IL-8, was similar in all patients. IL-8 mRNA levels were significantly decreased in the colonic mucosa of nicotine-treated patients who improved (p = 0.04). IL-8 mRNA values were similar before and after treatment in nonresponding nicotine-treated patients and in all placebo-treated patients. Mucin gene expression was similar in all patient groups. Beneficial effects of transdermal nicotine in active UC may result from decrease of IL-8 expression at the transcriptional level. Transdermal nicotine has no effect on mucin gene transcription.

Topics: Administration, Cutaneous; Adult; Blotting, Northern; Colitis, Ulcerative; Colon; Cytokines; Double-Blind Method; Female; Gene Expression; Humans; Interleukin-8; Intestinal Mucosa; Male; Mucins; Nicotine; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

The inflammatory activity of colonic mucosal lesions may be stimulated by intraluminal bacteria. Our aim was to investigate whether administration of broad-spectrum antibiotics decreases inflammatory activity in ulcerative colitis. To this end, we performed a randomized, 5-day study with either oral enterically coated amoxicillin-clavulanic acid (1 g + 250 mg, t.i.d.); i.v. methylprednisolone (40 mg/day) and oral placebo (t.i.d.); or both i.v. methylprednisolone and oral amoxicillin-clavulanic acid as above, in 30 patients with clinically active ulcerative colitis. Before and after 5 days of treatment, intestinal inflammation was assessed by the quantification of mucosal release of eicosanoids and interleukin-8 by rectal dialysis in each patient. Breath H2 excretion after oral lactulose was determined as an index of metabolic activity of colonic flora. The total release of (IL-8) interleukin-8 and eicosanoids significantly decreased in patients treated with antibiotic or steroids and antibiotic. Antibiotic treatment, but not steroids, markedly inhibited breath H2 excretion. In conclusion, short-term treatment with enteric-coated amoxicillin-clavulanic acid decreases the intraluminal release of IL-8 and other inflammatory mediators.

Topics: Adolescent; Adult; Aged; Amoxicillin-Potassium Clavulanate Combination; Anti-Inflammatory Agents; Colitis, Ulcerative; Drug Therapy, Combination; Eicosanoids; Female; Humans; Interleukin-8; Male; Methylprednisolone; Middle Aged; Tablets, Enteric-Coated

Ulcerative Colitis (UC) is a chronic nonspecific inflammatory disease of the colon and rectum. Fructus Mume (FM) and Rhizoma Coptidis (RC) exert effects on inflammatory and immune diseases. We evaluated the hypothesis of the FM and RC (FM-RC) herb pair remedy in alleviating dextran sulfate sodium (DSS)-induced colitis, through network pharmacology-based analyses, molecular docking, and experimental validation.. The Traditional Chinese medicine systematic pharmacology analysis platform(TCMSP) and Swiss database were used to predict potential targets of FM-RC and the GeneCards database was utilized to collect UC genes. Cytoscape software was used to construct and analyze the networks, and DAVID was utilized to perform enrichment analysis. AutoDock software was used to dock the core chemical components of the FM-RC herb pair with key UC targets. Animal experiments were performed to validate the prediction results and general conditions and body weight were observed. Pathological changes in colon tissue were observed by staining with hematoxylin and eosin. The levels of TNF-α, IL-8, IL-17, and IL-4 in serum and colon tissue were detected by ELISA.. Eighteen effective components of the herb couple were screened, and their potential therapeutic targets in the treatment of UC were acquired from 110 overlapped targets. GO and KEGG analyses revealed that these targets were highly correlated with protein autophosphorylation, plasma membrane, ATP binding, cancer pathways, the PI3K-AKt signaling pathway, and the Rap1 signaling pathway. Molecular docking established the core protein interactions with compounds having a docking energy < 0 kJ·mol. Collectively, FM-RC herb pair administration alleviated UC. These beneficial effects targeted MAPK1 signaling related to inflammation and immunity, which provided a basis for a better understanding of FM-RC in the treatment of UC.

Topics: Animals; Antineoplastic Agents; Colitis, Ulcerative; Drugs, Chinese Herbal; Inflammation; Interleukin-17; Interleukin-4; Interleukin-8; Molecular Docking Simulation; Phosphatidylinositol 3-Kinases; Rats; Tumor Necrosis Factor-alpha

Topics: Adenocarcinoma; Anti-Inflammatory Agents; Caco-2 Cells; Colitis, Ulcerative; Colonic Neoplasms; Cyclooxygenase 2; Dioscorea; Gene Silencing; Humans; Interleukin-8; Molecular Docking Simulation; NF-kappa B; Signal Transduction; Tumor Necrosis Factor-alpha

Complete endoscopic mucosal healing is defined as a Mayo endoscopic subscore of 0. Some patients diagnosed with a Mayo endoscopic subscore 0 may present with subsequent clinical relapse. Here, we aimed to demonstrate mucosal cytokine profile as a predictor of clinical relapse in ulcerative colitis patients with a Mayo endoscopic subscore of 0 as a marker of mucosal healing.. We conducted prospective observational pilot study to examine the relationship between mucosal cytokine expression and subsequent relapse of UC patients diagnosed with a Mayo endoscopic subscore of 0. We enrolled 55 patients, and expression of cytokines tumor necrosis factor-α, interferon γ, interleukin-1β, interleukin-2, interleukin-4, interleukin-5, interleukin-6, interleukin-7, interleukin-8, interleukin-9, interleukin-10, interleukin-12, interleukin-13, interleukin-15, interleukin-17A, interleukin-17F, interleukin-18, interleukin-21, interleukin-22, interleukin-23, interleukin-27, and interleukin-33 was measured by quantitative real-time PCR using rectal mucosa biopsy materials. Cytokine expression levels were compared between patients who relapsed between March 1, 2016, and March 30, 2020, of the study period and those who remained in remission.. Ten cytokines, including interleukin-2, interleukin-4, interleukin-8, interleukin-10, interleukin-12, interleukin-15, interleukin-17A, interleukin-21, interleukin-23, and interleukin-33, were significantly elevated in patients with subsequent relapse compared with those who remained in remission. Interleukin-8 expression was the most useful predictor.. In the rectal mucosa of ulcerative colitis patients with Mayo endoscopic subscore 0, levels of several cytokines were elevated in cases of subsequent relapse. Among these, interleukin-8 expression was the most useful for predicting relapse.

Topics: Colitis, Ulcerative; Colonoscopy; Humans; Interleukin-10; Interleukin-12; Interleukin-15; Interleukin-17; Interleukin-2; Interleukin-23; Interleukin-33; Interleukin-4; Interleukin-8; Intestinal Mucosa; Prospective Studies; Recurrence; Severity of Illness Index

Celiac disease (CeD) and inflammatory bowel disease (IBD) are accompanied by impaired immune responses. To study the immune regulation of these diseases, we evaluated the expression levels of pro-inflammatory (IL-8 and IL-17 A) and anti-inflammatory (IL-10) cytokines in intestinal biopsy specimens of CeD and IBD patients in comparison to healthy subjects.. Intestinal biopsies were collected from 33 patients with IBD, 47 patients with CeD, and 20 healthy individuals. Total RNA was extracted and mRNA expression levels of IL-8, IL-17 A and IL-10 were assessed by qPCR. P-value < 0.05 was considered statistically significant. The expression levels of IL-8 and IL-17 A were higher in biopsies of IBD (UC and CD) and CeD patients compared to the control group (P < 0.05). IBD patients (UC and CD) had higher IL-8 intestinal level than CeD patients (P < 0.0001 and P = 0.0007, respectively). The expression of IL-10 was significantly down-regulated in intestinal biopsies of CeD and IBD patients compared with controls (P < 0.001). In addition, the expression level of this cytokine was significantly lower in IBD patients (P < 0.001 for UC patients and P < 0.0001 for CD patients) than CeD group.. The three selected pro- and anti-inflammatory cytokines showed a similar expression pattern in both IBD and CeD patients. As IBD and CeD are immune-mediated disorders and are accompanied by inflammatory events, the understanding of the similarities and differences among them can help researchers to find out useful candidate therapeutic protocols. We suggest that larger cohort studies be organized to achieve more insights into this regulation.

Topics: Colitis, Ulcerative; Cytokines; Gene Expression; Humans; Inflammatory Bowel Diseases; Interleukin-10; Interleukin-17; Interleukin-8; Intestinal Mucosa

Recent progress in ulcerative colitis (UC) treatment has been remarkable, and various medications have been applied. However, some patients with UC are refractory to treatment and convert to surgery.. To investigate the role of colonic mucosal Wnt-5a expression in the pathogenesis of UC and the effect of bioactive Wnt-5a peptide on colitis in mice.. Wnt-5a peptide was intraperitoneally administered to mice every day from the beginning of dextran sulfate sodium (DSS) treatment. The severity of colitis was evaluated based on body weight change, colonic length, and histological scores. Colonic mucosal TNF-α and KC mRNA expression levels were measured. This study included 70 patients with UC in clinical remission. Wnt-5a, TNFα, and IL-8 mRNA expression in the rectal mucosa were measured by quantitative real-time polymerase chain reaction using biopsy materials. Wnt-5a mRNA expression levels were compared between patients who relapsed and those in remission. We examined the correlation of Wnt-5a expression with TNF-α and IL-8 expression.. Wnt-5a peptide significantly attenuated the severity of DSS-induced colitis. Moreover, mucosal TNF-α and KC mRNA expression were significantly suppressed by Wnt-5a peptide treatment. Wnt-5a mRNA levels were significantly lower in patients with subsequent relapse than in those who remained in remission. Mucosal Wnt-5a was inversely correlated with TNF α and IL-8 expression.. Wnt-5a peptide suppressed colitis in mice, and decreased Wnt-5a expression was strongly associated with relapse in patients with UC. Wnt-5a may have an inhibitory effect on mucosal inflammation in UC, and Wnt-5a peptide could be a new therapeutic strategy.

Topics: Animals; Colitis; Colitis, Ulcerative; Colon; Dextran Sulfate; Inflammation; Interleukin-8; Intestinal Mucosa; Mice; Recurrence; RNA, Messenger; Tumor Necrosis Factor-alpha

To explore the immune change of lung injury of Ulcerative colitis (UC) by observing the changes of inherent immunity and adaptive immunity of the lung and bowel in UC rat models after the treatment of Sodium Houttuyfonate combined with Matrine.. UC rat models were established with the mucous membrane of colon allergize combined with TNBS-alcohol enteroclysis for 1 week and 5 weeks. 1-week experimental rats were divided into normal group and model group, 5/each group. 5-weeks experimental rats were divided into normal group, model group, Sodium Houttuyfonate (2.9mg/ml) combined with Matrine (1.47mg/ml), and positive control sulfasalazine (10mg/ml), 5/each group. All rats were administered by gavage for 5 weeks. The histopathological and fibrotic changes in the lung and bowel were observed, and the expressions of Tumor Necrosis Factor (TNF)- α, interleukin (IL)-8 in the lung, bowel, and serum were detected by radio-immunity and immunohistochemistry, and the mRNA expressions of Toll-like receptor (TLR)-4, nuclear factor kappa (NF-κB), Macrophage migration inhibitory factor (MIF), Mucosal addressing cell adhesion molecule-1 (MadCAM1) and Pulmonary surfactant protein-A (SP-A) in the lung and bowel were detected by Real time-PCR.. Compared with the normal group, the model rats had significant histopathological and fibrotic changes both in the lung and bowel, and all treatment groups were improved. After treatment, TLR4, IL-8, MIF, and TNF-α in the lung decreased (P<0.05); NF-KB, IL-8, and MIF in the bowel increased (P<0.05); MadCAM1 both in lung and bowel decreased (P<0.05); SP-A decreased in bowel and increased in the lung (P<0.05).. The cause of lung injury in this model was found to be related to inherent immunity and adaptive immunity, while the cause of bowel injury in this model was found to be mainly related to adaptive immunity. Sodium Houttuyfonate combined with Matrine could improve bowel and lung injury.

Topics: Alkaloids; Alkanes; Animals; Colitis, Ulcerative; Interleukin-8; Lung; Lung Injury; Matrines; NF-kappa B; Quinolizines; Rats; Signal Transduction; Sulfites; Tumor Necrosis Factor-alpha

At present, the most direct method to evaluate mucosal healing in patients with ulcerative colitis (UC) is endoscopy, but it is costly and invasive. Therefore, it is necessary to find a biomarker with low cost, easy access, high sensitivity and specificity as an indicator of UC activity. This study aimed to examine the level of platelet to lymphocyte ratio (PLR) and interleukin-8 (IL-8) in UC patients and evaluate their roles in differential diagnosis and disease activity assessment.. A retrospective study involving 130 UC patients and 141 irritable bowel syndrome (IBS) patients was performed. The UC patients were divided into remission group and active group according to the Modified Mayo score. The receiver operating characteristic curve (ROC) analysis was performed to determine the optimal cutoff value of PLR and IL-8 in differential diagnosis between UC patients and IBS patients.. The levels of WBC, PLR, and IL-8 in UC patients were higher than those compared with IBS controls. The optimal cutoff to differentiate UC and IBS patients was 6.76 109/L, 114.70, and 19.42 pg/mL for WBC, PLR, and IL-8, respectively (sensitivity, 36.9% vs. 83.8% vs. 72.3%; specificity, 83.0% vs. 65.2% vs. 94.3%; AUC, 0.601 vs. 0.815 vs. 0.859). IL-8 had the highest AUC and specificity. Among 130 patients, 75 patients (57.6%) had mucosal inflammation. The cutoff value of IL-8 for predicting disease severity of UC patients was 22.21 pg/mL (AUC: 0.861). The sensitivity, specificity, and Youden index of IL-8 for predicting severe UC were 92.0%, 81.8%, and 0.702, respectively.. PLR and IL-8 showed great performance in distinguishing UC from IBS patients. Moreover, elevated IL-8 level indicated mucosal inflammation, reflecting disease severity in UC patients.

Topics: Biomarkers; Colitis, Ulcerative; Diagnosis, Differential; Humans; Inflammation; Interleukin-8; Irritable Bowel Syndrome; Lymphocytes; Retrospective Studies

The function of interleukin-22 (IL-22) in intestinal barrier homeostasis remains controversial. Here, we map the transcriptional landscape regulated by IL-22 in human colonic epithelial organoids and evaluate the biological, functional and clinical significance of the IL-22 mediated pathways in ulcerative colitis (UC). We show that IL-22 regulated pro-inflammatory pathways are involved in microbial recognition, cancer and immune cell chemotaxis; most prominently those involving CXCR2

Topics: Chemokines, CXC; Colitis, Ulcerative; Humans; Interleukin-22; Interleukin-8; Interleukins; Neutrophil Infiltration; Neutrophils; Receptors, Interleukin-8B; Ustekinumab

Dilodendron bipinnatum (Sapindaceae) stem bark decoction and macerate were used to treat uterine inflammation, pain in general, dermatitis and bone fractures. These homemade preparations also have diuretic, stimulant, expectorants and sedative effects and are effective in treating worm infections in the Brazilian Pantanal population. Our previous research confirmed the anti-inflammatory activity of the hydroethanolic extract of inner stem bark of D. bipinnatum (HEDb).. This work aimed to investigate the efficacy of HEDb in ameliorating experimental colitis in rats and to elucidate the possible mechanisms involved in the anti-ulcerative colitis properties of HEDb in rats and Caco-2 cell line.. The effects on cell viability, IL-8 and TNF-α in human colon adenocarcinoma (Caco-2) were determined by flow cytometer and ELISA. Wistar rats (n = 6-7) were orally gavaged with, vehicle (0.9% saline), HEDb at doses of 20, 100 or 500 mg/kg, or mesalazine at a dose of 500 mg/kg, at 48, 24 and 1 h prior to the administration of trinitrobenzene sulfonic acid via rectal administration to induce colitis. The anti-inflammatory effects of HEDb were assessed macroscopically, by myeloperoxidase (MPO) activity and for glutathione (GSH) concentration in the colon. Additionally, colonic histopathological analyses of UC severity were conducted by different staining methods (H&E, PAS and toluidine blue). Pro-inflammatory cytokines TNF-α and IL-1β were quantified in colonic tissue by ELISA and colonic expressions of COX-2 and IL-17 were analyzed by western blotting.. HEDb was shown to be non-cytotoxic with mean viability of 80% in Caco-2 cells. HEDb pre-treatments of 1, 5 or 20 μg/mL significantly reduced TNF-α production in Caco-2 cells by 21.8% (p < 0.05), 60.5 and 82.1% (p < 0.001) respectively following LPS treatment compared to LPS alone. However, no change in IL-8 production was observed. HEDb pre-treatment of rats subjected to TNBS significantly (p < 0.001) reduced colonic lesion score. Higher doses (100 and 500 mg/kg) caused a sharp downregulation of haemorrhagic damage, leukocyte infiltration, edema and restoration of mucus production. Moreover, mast cell degranulation was inhibited. Colonic MPO activity was reduced following all doses of HEDb, reaching 51.1% ± 1.51 (p < 0.05) with the highest dose. GSH concentration was restored by 58% and 70% following 100 and 500 mg/kg of HEDb, respectively. The oral treatment of HEDb at doses 20, 100 and 500 mg/kg decreased the concentrations of TNF-α and IL-1β at all doses in comparison to vehicle treated control. In addition, HEDb inhibited the COX-2 and IL-17 expressions with maximal effect at 500 mg/kg (60.3% and 65% respectively; p < 0.001). In all trials, the effect of HEDb at all doses being 20, 100 and 500 mg/kg was statistically comparable to mesalazine (500 mg/kg).. HEDb reduces colonic damage in the TNBS colitis model and relieves oxidative and inflammatory events, at least in part, by increasing mucus production, reducing leukocyte migration and reducing TNF-α (in vivo and in vitro), IL-1β, IL-17 and COX-2 expression. Therefore, HEDb requires further investigation as a candidate for treating IBD.

Topics: Animals; Antioxidants; Brazil; Caco-2 Cells; Cell Survival; Colitis, Ulcerative; Cyclooxygenase 2; Disease Models, Animal; Edema; Glutathione; Humans; Interleukin-17; Interleukin-1beta; Interleukin-8; Lipopolysaccharides; Mast Cells; Mucus; Peroxidase; Plant Bark; Plant Extracts; Rats, Wistar; Sapindaceae; Trinitrobenzenesulfonic Acid; Tumor Necrosis Factor-alpha

BACKGROUND This bioinformatics study aimed to identify differentially expressed genes (DEGs) and protein-protein interaction (PPI) networks associated with functional pathways in ulcerative colitis based on 3 Gene Expression Omnibus (GEO) datasets. MATERIAL AND METHODS The GSE87466, GSE75214, and GSE48958 MINiML formatted family files were downloaded from the GEO database. DEGs were identified from the 3 datasets, and volcano maps and heat maps were drawn after R language standardization and analysis, respectively. Venn diagram software was used to identify common DEGs. PPI analysis of common DEGs was performed using the Search Tool for the Retrieval of Interacting Genes. Gene modules and hub genes were visualized in the PPI network using Cytoscape. Enrichment analysis was performed for all common DEGs, module genes, and hub genes. RESULTS A total of 90 DEGs were selected, which included 3 functional modules and 1 hub gene module. CXCL8 module genes were mainly enriched in cytokine-mediated signaling pathways and interleukin (IL)-10 signaling. CCL20 module genes were mainly enriched in the IL-17 signaling pathway and cellular response to IL-1. Hub gene modules mainly involved IL-10, IL-4, and IL-13 signaling pathways. CXCL8, CXCL1, and IL-1ß were the top 3 hub genes and were mainly involved in IL-10 signaling. CONCLUSIONS Bioinformatics analysis using 3 GEO datasets identified CXCL8, CXCL1, and IL-1ß, which are involved in IL-10 signaling, as the top 3 hub genes in ulcerative colitis. The findings from this study remain to be validated, but they may contribute to the further understanding of the pathogenesis of ulcerative colitis.

Topics: Chemokine CXCL1; Colitis, Ulcerative; Computational Biology; Gene Regulatory Networks; Humans; Interleukin-1beta; Interleukin-8; Protein Interaction Maps; Transcriptome

Pregnancy may affect the disease course of IBD. Both pregnancy and IBD are associated with altered immunology and intestinal microbiology. However, to what extent immunological and microbial profiles are affected by pregnancy in patients with IBD remains unclear.. Faecal and serum samples were collected from 46 IBD patients (31 Crohn's disease (CD) and 15 UC) and 179 healthy controls during first, second and third trimester of pregnancy, and prepregnancy and postpartum for patients with IBD. Peripheral blood cytokine profiles were determined by ELISA, and microbiome analysis was performed by sequencing the V4 region of the bacterial 16S rRNA gene.. Proinflammatory serum cytokine levels in patients with IBD decrease significantly on conception. Reduced interleukin (IL)-10 and IL-5 levels but increased IL-8 and interferon (IFN)γ levels compared with healthy controls were seen throughout pregnancy, but cytokine patterns remained stable during gestation. Microbial diversity in pregnant patients with IBD was reduced compared with that in healthy women, and significant differences existed between patients with UC and CD in early pregnancy. However, these microbial differences were no longer present during middle and late pregnancy. Dynamic modelling showed considerable interaction between cytokine and microbial composition.. Serum proinflammatory cytokine levels markedly improve on conception in pregnant patients with IBD, and intestinal microbiome diversity of patients with IBD normalises during middle and late pregnancy. We thus conclude that pregnancy is safe and even potentially beneficial for patients with IBD.

Topics: Adult; Case-Control Studies; Colitis, Ulcerative; Crohn Disease; Cytokines; Feces; Female; Gastrointestinal Microbiome; Humans; Interferon-gamma; Interleukin-10; Interleukin-5; Interleukin-8; Pregnancy; Pregnancy Complications; Pregnancy Trimesters

CD14+ mononuclear phagocytes [MNPs] and T cells infiltrate colon in ulcerative colitis [UC]. Here we investigated how CD14+ MNPs and the cytokines they produce shape the colonic effector T cell profile.. Colonic or mesenteric lymph node [mLNs] CD4+ T cells isolated from UC or Crohn's disease [CD] patients were stimulated with cytokines or autologous CD14+ MNPs. Cytokine expression was assessed by intracytoplasmic staining and multiplex ELISA. Unsupervised phenotypic multicolour analysis of colonic CD14+ MNPs was performed using the FlowSOM algorithm.. Among CD14+CD64+HLA-DR+SIRPα + MNPs, only the pro-inflammatory cytokine-producing CD163- subpopulation accumulated in inflamed UC colon and promoted mucosal IL-1β-dependent Th17, Th17/Th1, Th17/Th22 but not Th1 responses. Unsupervised phenotypic analysis of CD14+CD64+ MNPs segregated CD163- monocyte-like cells and CD163+ macrophages. Unexpectedly, IL-12, IL-1β and CD163-, but not CD163+, cells induced IL-8 expression in colonic CD4+ T cells, which co-expressed IFN-γ and/or IL-17 in UC and not CD. The CD163- monocyte-like cells increased the frequency of IL-8+IL-17+/-IFN-γ +/- T cells through IL-1β and IL-12. Finally, colonic IL-8+ T cells co-expressing GM-CSF, TNF-α and IL-6 were detected ex vivo and, promoted by IL-12 in the mucosa and mLNs in UC only.. Our findings established a link between monocyte-like CD163- MNPs, IL-12, IL-1β and the detection of colonic memory IL-8-producing CD4+ T cells, which might all contribute to the pathogenesis of UC.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Case-Control Studies; CD4-Positive T-Lymphocytes; Colitis, Ulcerative; Crohn Disease; Female; Humans; Interleukin-12; Interleukin-8; Intestinal Mucosa; Lipopolysaccharide Receptors; Male; Middle Aged; Young Adult

Inflammatory bowel disease is a chronic, relapsing condition with two subtypes, Crohn's disease (CD) and ulcerative colitis (UC). Genome-wide association studies (GWASs) in UC implicate a FCGR2A variant that alters the binding affinity of the antibody receptor it encodes, FcγRIIA, for immunoglobulin G (IgG). Here, we aimed to understand the mechanisms whereby changes in FcγRIIA affinity would affect inflammation in an IgA-dominated organ. We found a profound induction of anti-commensal IgG and a concomitant increase in activating FcγR signaling in the colonic mucosa of UC patients. Commensal-IgG immune complexes engaged gut-resident FcγR-expressing macrophages, inducing NLRP3- and reactive-oxygen-species-dependent production of interleukin-1β (IL-1β) and neutrophil-recruiting chemokines. These responses were modulated by the FCGR2A genotype. In vivo manipulation of macrophage FcγR signal strength in a mouse model of UC determined the magnitude of intestinal inflammation and IL-1β-dependent type 17 immunity. The identification of an important contribution of IgG-FcγR-dependent inflammation to UC has therapeutic implications.

Topics: Animals; Antibodies, Bacterial; Colitis; Colitis, Ulcerative; Dextran Sulfate; Gastrointestinal Microbiome; Gene Expression Regulation; Genotype; Humans; Immunoglobulin G; Inflammation; Interleukin-1beta; Interleukin-8; Intestinal Mucosa; Macrophages; Mice; Phagocytes; Reactive Oxygen Species; Receptors, IgG; RNA, Messenger; Th17 Cells

Inflammatory bowel disease (IBD) is a common disorder of chronic intestinal inflammation that can be caused by the disruption of intestinal immune homeostasis.. We aimed to evaluate the role of enhancer of zeste homolog 2 (EZH2) in the inflammatory response and explore the association between EZH2 and necroptosis in human epithelial colorectal adenocarcinoma cell lines.. In both in vitro and in vivo models, expression of EZH2 in intestinal tissues was verified by histology. The expression of inflammatory cytokines in cell lines treated with EZH2 siRNA with or without stimulus was analyzed by quantitative real-time polymerase chain reaction. An intestinal necroptosis cell model was established to elucidate whether EZH2 is involved in necroptosis.. Our present data indicated that EZH2 expression was decreased in in vitro and in vivo models and in patients with inflammatory bowel disease. EZH2 downregulation increased the expression of inflammatory factors, including TNF-α, IL-8, IL-17, CCL5, and CCL20 in a Caco-2 cell model. The JNK pathway was activated with the reduction of EZH2. In the necroptosis model, downregulation of EZH2 was detected with the upregulation of necroptotic markers RIP1 and RIP3. In addition, EZH2 knockdown with siRNA increased p-JNK and p-c-Jun.. Our data suggest that EZH2 plays an important role in the development of intestinal inflammation and necroptosis. Hence, EZH2 could be a potential therapeutic target for IBD.

Topics: Animals; Caco-2 Cells; Chemokine CCL20; Chemokine CCL5; Colitis; Colitis, Ulcerative; Crohn Disease; Dextran Sulfate; Down-Regulation; Enhancer of Zeste Homolog 2 Protein; Gene Knockdown Techniques; Humans; In Vitro Techniques; Inflammation; Inflammatory Bowel Diseases; Interleukin-17; Interleukin-8; Intestinal Mucosa; JNK Mitogen-Activated Protein Kinases; MAP Kinase Signaling System; Mice; Necroptosis; Nuclear Pore Complex Proteins; Phosphoproteins; Proto-Oncogene Proteins c-jun; Real-Time Polymerase Chain Reaction; Receptor-Interacting Protein Serine-Threonine Kinases; RNA-Binding Proteins; Trinitrobenzenesulfonic Acid; Tumor Necrosis Factor-alpha

Inflammatory bowel disease (IBD) remains a major health challenge due in part to unsafe and limited treatment options, hence there is the need for alternatives. CXCL8/interleukin 8 (IL-8) is elevated in inflammation, and binds preferentially to G protein-couple receptors (GPCRs) CXCR1/2 of the CXC chemokine family to initiate cascades of downstream inflammatory signals. A mutant CXCL8 protein, CXCL8(3-72)K11R/G31P (G31P), competitively and selectively binds to CXCR1/2, making CXCL8 redundant. We explore the therapeutic potential of G31P in dextran sulfate sodium (DSS) induced ulcerative colitis (UC), and the corresponding effect if G31P treatment is augmented with Lactobacillus acidophilus (LACT). The treatment options administered significantly reduced TNF-α, IFN-γ, IL-1β, IL-6, and IL-8, but maintained elevated levels of IL-10. CD68 and F4/80 expressions were down-regulated and showed restricted infiltration to inflamed colon, while IL-17F levels were insignificantly different from the DSS treated mice. Also, we observed up-regulation of IL-17A in G31P + LACT but not G31P treated mice if compared with Control group. The treatments ameliorated colonic fibrosis by reducing VEGF, TGF-β, MMP-2 and MMP-9. In addition, we observed elevated levels of E-cadherin, and marginal up-regulation of occludin, suggesting the role of the treatments in regulating tight intestinal junction and adherence proteins. Mechanism-wise, G31P interferes with AKT and ERK signaling pathways. Our study suggests that G31P confers protection in IBD, particularly UC, and when G31P treatment is augmented with Lactobacillus acidophilus, the protection is variably enhanced.

Topics: Animals; Colitis, Ulcerative; Colon; Cytokines; Dextran Sulfate; Disease Models, Animal; Early Growth Response Protein 1; Fibrosis; Inflammation; Inflammation Mediators; Interleukin-8; Male; MAP Kinase Signaling System; Mice, Inbred C57BL; Mutant Proteins; Probiotics; Proto-Oncogene Proteins c-akt; Tight Junction Proteins

Ulcerative colitis (UC) is a kind of chronic inflammatory bowel diseases that seriously endangers human health. The pathogenesis of UC is closely related to the intestinal immune response. Cytokines exert a key role in the regulation of intestinal inflammatory and immune responses. Abnormalities in the function and quantity of various cytokines or imbalance of inflammatory factors and immune factors would lead to UC development. We aimed to investigate whether Kruppel-like transcription factor 2 (KFL2) participates in the development of ulcerative colitis by regulating inflammation, so as to provide a new direction for the clinical treatment.. 40 UC patients were enrolled in this study, including 20 patients with mild ulcerative colitis (MUC) and 20 with severe ulcerative colitis (SUC). 20 normal end of intestinal tissues surgically resected from patients with colorectal cancer in the same period were selected as the control group. Hematoxylin-eosin (HE) staining was used to detect the inflammatory infiltration of intestinal mucosa tissues. Expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-α) in peripheral blood mononuclear cells (PBMCs) of each group were detected by qRT-PCR (quantitative Real-Time Polymerase Chain Reaction). Immunohistochemistry was performed to observe the infiltration of IL-6 and TNF-α in intestinal mucosal tissues. Protein and mRNA levels of KLF2 in PBMCs of each group were detected by Western blot and qRT-PCR, respectively. The relationship between the mRNA level of KLF2 in PBMCs and expressions of IL-6, IL-8, IL-10, TNF-α were analyzed using qRT-PCR.. Inflammatory cells and cytokines were infiltrated in the intestinal mucosa of UC patients. IL-6, IL-8, IL-10, and TNF-α were overexpressed in PBMCs of UC patients than those of controls. Protein and mRNA levels of KLF2 in PBMCs of UC patients were remarkably lower than those of controls, which were more significant in SUC patients. Meanwhile, KLF2 was closely related to expressions of IL-6, IL-8, IL-10, and TNF-α in PBMCs of UC patients.. KLF2 was downregulated in PBMCs of UC patients than that of normal controls, which participated in the inflammatory response of UC by regulating expressions of IL-6, IL-8, IL-10, and TNF-α. KLF2 may suggest new treatments for ulcerative colitis.

Topics: Adult; Aged; Case-Control Studies; Colitis, Ulcerative; Cytokines; Female; Humans; Inflammation; Interleukin-10; Interleukin-6; Interleukin-8; Intestinal Mucosa; Intestines; Kruppel-Like Transcription Factors; Leukocytes, Mononuclear; Male; Middle Aged; Severity of Illness Index; Tumor Necrosis Factor-alpha

Ulcerative colitis (UC) is a chronic inflammatory disease and its involvement area in colon is influenced by a complex network of gene interactions. We analyzed the weighted gene co-expression networks in microarray dataset from colonic mucosa of patients with UC and identified one gene co-expression module that was highly associated with the progression of involved area in UC colon (Pearson coefficient=0.81, P<0.0001). In total, 523 hub genes in this module were found to be involved in immune system process after enrichment analysis in Gene Ontology. By the STRING and Cytoscape analysis, we observed that interleukin-8 (IL-8) and matrix metalloproteinase-9 (MMP-9) were centered in the network of hub genes. We then detected the expression of IL-8 and MMP-9 in mucosa from left-sided colon of patients using quantitative PCR and immunofluorescence assay respectively. Both quantitative PCR and immunofluorescence assay revealed the expression levels of IL-8 and MMP-9 were significantly different among the healthy controls, left-sided colitis group and pancolitis group (P<0.05). IL-8 and MMP-9 were detected with an enhanced expression in pancolitis as compared with leftsided colitis and healthy controls, respectively (P<0.05). This study demonstrates that immune system process is indispensable in the progression of disease in colon, and identifies that IL-8 and MMP-9 play potential critical roles for the progression.

Topics: Case-Control Studies; Cluster Analysis; Colitis, Ulcerative; Colon; Demography; Disease Progression; Female; Gene Expression Regulation; Gene Ontology; Gene Regulatory Networks; Humans; Interleukin-8; Intestinal Mucosa; Male; Matrix Metalloproteinase 9; Middle Aged; RNA, Messenger; Transcriptome

Cytosolic pattern recognition receptors play key roles in innate immune response. Nucleotide binding and oligomerisation domain containing protein 1 (NOD1) belonging to the Nod-like receptor C (NLRC) sub-family of Nod-like receptors (NLRs) is important for detection and clearance of intra-cellular Gram negative bacteria. NOD1 is involved in activation of pro-inflammatory pathways. Limited structural data is available for NOD1. Using different templates for each domain of NOD1, we determined the full-length homology model of NOD1. ADP binding amino acids within the nucleotide binding domain (NBD) of NOD1 were also predicted. Key residues in inter-domain interaction were identified by sequence comparison with Oryctolagus cuniculus NOD2, a related protein. Interactions between NBD and winged helix domain (WHD) were found to be conserved in NOD1. Functional and structural effect of single nucleotide polymorphisms within the NOD1 NBD domain associated with susceptibility risk to Ulcerative colitis (UC), an inflammatory disorder of the colon was evaluated by in silico studies. Mutations W219R and L349P were predicted to be damaging and disease associated by prediction programs SIFT, PolyPhen2, PANTHER, SNP&GO, PhD SNP and SNAP2. We further validated the effect of W219R and L349P mutation on NOD1 function in vitro. Elevated mRNA expression of pro-inflammatory cytokines IL8 and IL-1β was seen as compared to the wild type NOD1 in intestinal epithelial cell line HT29 when stimulated with NOD1 ligand. Thus, these mutations may indeed have a bearing on pathogenesis of inflammation during UC.

Topics: Colitis, Ulcerative; Databases, Genetic; Exons; HT29 Cells; Humans; Interleukin-1beta; Interleukin-8; Mutation; Nod1 Signaling Adaptor Protein; Polymorphism, Single Nucleotide; Signal Transduction

Type 2 autoimmune pancreatitis (type 2 AIP) develops in isolation or sometimes in association with ulcerative colitis. Its diagnosis requires the histologic confirmation of granulocytic epithelial lesions (GELs) with no diagnostic biomarker currently available. This study aimed to elucidate the tissue expression of cytokines and their diagnostic value in this condition. In quantitative polymerase chain reaction for multiple cytokines using tissue-derived mRNA, the expression level of interleukin (IL)-8 was markedly higher in type 2 AIP than in type 1 AIP (P<0.001). In immunostaining, IL-8 expression was detected in the ductal/ductular epithelium (11/13; 85%) and infiltrating neutrophils or lymphocytes (12/12; 100%) in type 2 AIP, but was almost entirely negative in type 1 AIP (n=13; both, P<0.001). Although obstructive pancreatitis adjacent to pancreatic cancers (peritumoral pancreatitis) exhibited IL-8 expression in the epithelium (3/12; 25%) and inflammatory cells (10/12; 83%), expression levels were significantly lower than those in type 2 AIP (P<0.001 and 0.020, respectively). The presence of either GELs or IL-8-positive epithelium discriminated type 2 AIP from type 1 AIP or obstructive pancreatitis with 92% sensitivity and 92% to 100% specificity. Furthermore, CD3/IL-8-coexpressing lymphocytes were almost restricted to type 2 AIP. Interestingly, a similar pattern of IL-8 expression was also observed in colonic biopsies of ulcerative colitis. In conclusion, the overexpression of IL-8 may underlie the development of GELs in type 2 AIP, and IL-8 immunostaining or IL-8/CD3 double staining may become an ancillary method for its diagnosis. The similar expression pattern of IL-8 in ulcerative colitis also suggests a pathogenetic link between the 2 conditions.

Topics: Adolescent; Adult; Autoimmune Diseases; Colitis, Ulcerative; Female; Granulocytes; Humans; Interleukin-8; Male; Middle Aged; Pancreatitis; Young Adult

Necroptosis is an inflammatory form of programmed cell death requiring receptor-interacting protein kinase 3 (RIP3) and mixed lineage kinase domain-like protein (MLKL).. The aim of this study is to examine in depth in vitro and ex vivo the contribution of necroptosis to intestinal inflammation.. In vitro: we used an intestinal cell line, HCT116RIP3, produced in our laboratory and overexpressing RIP3. Ex vivo: intestinal mucosal biopsies were taken from patients with inflammatory bowel disease (IBD) (20 with Crohn's disease; 20 with ulcerative colitis) and from 20 controls.. RIP3-induced necroptosis triggers MLKL activation, increases cytokine/alarmin expression (IL-8, IL-1β, IL-33, HMGB1), NF-kBp65 translocation and NALP3 inflammasome assembly. It also affects membrane permeability by altering cell-cell junctional proteins (E-cadherin, Occludin, Zonulin-1). Targeting necroptosis through Necrostatin-1 significantly reduces intestinal inflammation in vitro and in cultured intestinal explants from IBD.. We show for the first time in vitro and ex vivo that RIP3-driven necroptosis seriously affects intestinal inflammation by increasing pMLKL, activating different cytokines and alarmins, and altering epithelial permeability. The inhibition of necroptosis causes a significant decrease of all these effects. These data strongly support the view that targeting necroptosis may represent a promising new option for the treatment of inflammatory enteropathies.

Topics: Adolescent; Amino Acid Chloromethyl Ketones; Apoptosis; Cadherins; Caspase 1; Cell Adhesion; Cell Membrane Permeability; Cell Survival; Child; Child, Preschool; Colitis, Ulcerative; Crohn Disease; Epithelial Cells; HCT116 Cells; HMGB1 Protein; Humans; Imidazoles; Indoles; Inflammasomes; Inflammation; Interleukin-1beta; Interleukin-8; Intestinal Mucosa; Necrosis; NLR Family, Pyrin Domain-Containing 3 Protein; Phosphorylation; Protein Kinases; Protein Transport; Receptor-Interacting Protein Serine-Threonine Kinases; RNA, Messenger; Transcription Factor RelA; Tumor Necrosis Factor-alpha

To investigate interleukin (IL)-26 expression in the inflamed mucosa of patients with inflammatory bowel disease (IBD) and the function of IL-26.. Human colonic subepithelial myofibroblasts (SEMFs) were isolated from colon tissue surgically resected. The expression of IL-26 protein and its receptor complex was analyzed by immunohistochemistry. The gene expression induced by IL-26 was evaluated by real-time polymerase chain reaction. Intracellular signaling pathways were evaluated by immunoblotting and specific small interfering (si) RNA transfection.. These results suggest that IL-26 plays a role in the pathophysiology of IBD through induction of inflammatory mediators.

Topics: Biopsy; Colitis, Ulcerative; Colon; Crohn Disease; Humans; Immunohistochemistry; Interleukin-6; Interleukin-8; Interleukins; Intestinal Mucosa; Mitogen-Activated Protein Kinases; Myofibroblasts; NF-kappa B; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Primary Cell Culture; Real-Time Polymerase Chain Reaction; RNA Interference; RNA, Messenger; RNA, Small Interfering; Signal Transduction; STAT1 Transcription Factor; STAT3 Transcription Factor; Transcription Factor AP-1; Up-Regulation

To investigate the efficacy and safety of stellate ganglion block for the treatment of patients with chronic ulcerative colitis.. A total of 120 randomly selected patients with chronic ulcerative colitis treated in Cangzhou Central Hospital from January 2014 to January 2016 were included in this study. These patients were divided into two groups: control group (. After treatment, clinical symptoms and disease activity were shown to be alleviated by endoscopy in both the control and experimental groups. However, patients in the control group did not have obvious abdominal pain relief. In addition, the degree of pain relief in the experimental group was statistically better than that in the control group (. The application of stellate ganglion block effectively improves treatment efficacy in chronic ulcerative colitis, relieves clinical symptoms in patients, and reduces the level of inflammatory factors. Furthermore, this approach also had a positive impact on the disease to a certain extent.

Topics: Administration, Oral; Adult; Anti-Inflammatory Agents, Non-Steroidal; Autonomic Nerve Block; Colitis, Ulcerative; Colonoscopy; Enzyme-Linked Immunosorbent Assay; Female; Humans; Injections, Spinal; Interleukin-8; Lidocaine; Male; Middle Aged; Stellate Ganglion; Sulfasalazine; Treatment Outcome

To investigate the therapeutic effect of Jianpi Qingchang decoction (JPQCD) on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice.. C57BL/c mice were injected intragastrically with 5% DSS instead of drinking water for 7 d, and their body weight, diarrhea severity and fecal bleeding were monitored, while the mice in the control group were treated with standard drinking water, without DSS. After 7 d, the DSS drinking water was changed to normal water and the DSS group continued with DSS water. The control and DSS groups were given normal saline by intragastric injection. The 5-aminosalicylic acid (5-ASA) group was treated orally with 5-ASA at a dose of 100 mg/kg daily. The JPQCD group was treated orally with JPQCD at a dose of 17.1 g/kg daily. On day 14, the colon length was measured, the colorectal histopathological damage score was assessed, and protein levels of interleukin (IL)-1β, IL-8 and tumor necrosis factor-alpha (TNF-α) in colon supernatants were measured by enzyme-linked immunosorbent assay. mRNA expression of IL-1β, IL-8, TNF-α and nuclear factor-kappa B (NF-κB) was detected by real-time quantitative polymerase chain reaction. Western blotting was used to detect the protein expression of NF-κB and inhibitor of kappa B.. Acute inflammation occurred in the mice administered DSS, including the symptoms of losing body weight, loose feces/watery diarrhea and presence of fecal blood; all these symptoms worsened at 7 d. The colons of mice treated with DSS were assessed by histological examination, and the results confirmed that acute inflammation had occurred, as evidenced by loss of colonic mucosa and chronic inflammatory cell infiltration, and these features extended into the deeper layer of the colon walls. The expression levels of IL-1β, IL-8 and TNF-α in the DSS group were higher than those in the control group (. Activation of the NF-κB signaling pathway is inhibited by JPQCD, which shows the potential mechanism by which JPQCD treats UC.

Topics: Animals; Colitis; Colitis, Ulcerative; Colon; Dextran Sulfate; Drugs, Chinese Herbal; Inflammation; Interleukin-1beta; Interleukin-8; Male; Mesalamine; Mice; Mice, Inbred C57BL; NF-kappa B; Signal Transduction; Tumor Necrosis Factor-alpha

Periostin is a matricellular protein that interacts with various integrin molecules on the cell surface. Although periostin is expressed in inflamed colonic mucosa, its role in the regulation of intestinal inflammation remains unclear. We investigated the role of periostin in intestinal inflammation using Postn-deficient (Postn-/-) mice. Intestinal epithelial cells (IECs) were transfected by Postn small interfering RNAs. Periostin expression was determined in colon tissue samples from ulcerative colitis (UC) patients. Oral administration of dextran sulfate sodium (DSS) or rectal administration of trinitrobenzene sulfonic acid, induced severe colitis in wild-type mice, but not in Postn-/- mice. Administration of recombinant periostin induced colitis in Postn-/- mice. The periostin neutralizing-antibody ameliorated the severity of colitis in DSS-treated wild-type mice. Silencing of Postn inhibited inteleukin (IL)-8 mRNA expression and NF-κB DNA-binding activity in IECs. Tumor necrosis factor (TNF)-α upregulated mRNA expression of Postn in IECs, and recombinant periostin strongly enhanced IL-8 expression in combination with TNF-α, which was suppressed by an antibody against integrin αv (CD51). Periostin and CD51 were expressed at significantly higher levels in UC patients than in controls. Periostin mediates intestinal inflammation through the activation of NF-κB signaling, which suggests that periostin is a potential therapeutic target for inflammatory bowel disease.

Topics: Acute Disease; Animals; Antibodies, Neutralizing; Cell Adhesion Molecules; Cell Line; Colitis, Ulcerative; Colon; Cytokines; Disease Models, Animal; Enterocytes; Gene Silencing; Humans; Inflammation; Integrin alphaV; Interleukin-8; Intestines; Male; Mice, Inbred C57BL; NF-kappa B; Protein Binding; Recombinant Proteins; Signal Transduction

Our recent study has demonstrated that medium chain triglycerides (MCT) and monounsaturated fatty acids potentiate the beneficial effects of fish oil on risk factors of cardiovascular disease. In the present study, we have investigated the influence of MCT or olive oil on the protective and mucosal healing ability of fish oil in ulcerative colitis using cell simulation and animal models. Caco-2 cells grown in medium chain fatty acids enriched medium has exaggerated t-butyl hydroperoxide induced cell damage, GSH depletion, and IL-1β induced IL-8 synthesis, compared to the cells grown in oleic acid & hydroxytyrosol (OT) enriched medium. Further, combined treatment of cells with eicosapentaenoic acid, docosahexaenoic acid, and OT has remarkably attenuated the cell damage, and IL-8 synthesis, compared to individual treatments. To evaluate the effect of these lipid formulations in vivo, adult Wistar rats were fed diet enriched with high amount of medium chain triglycerides (MCT), virgin olive oil, or their combination with fish oil. Colitis was induced in rats using dextran sulfate sodium (DSS) for 7days followed by 10-days of recovery period. Rats of MCT group exhibit severe disease activity, higher levels of inflammatory cytokines in the colon compared to the olive oil group. Furthermore, there was persistent body weight loss, loose stools, higher levels of inflammatory cytokines in the rats of MCT group, even after DSS was withdrawn from drinking water. Conversely, fish oil has remarkably attenuated the DSS induced alterations in both MCT and olive oil diet groups with significantly greater effect in the olive oil group. Thus, MCT increase the susceptibility to colitis through oxidative damage and IL-8 synthesis in intestinal epithelial cells. The beneficial effects of virgin olive oil could be partially attributed to hydroxytyrosol. Combined treatment of hydroxytyrosol, oleic acid and n-3 fatty acids exhibit huge therapeutic benefits in colitis.

Topics: Animals; Caco-2 Cells; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Drug Synergism; Drug Therapy, Combination; Fatty Acids, Omega-3; Fish Oils; Humans; Interleukin-8; Male; Oleic Acid; Oxidative Stress; Phenylethyl Alcohol; Rats; Rats, Wistar

Kinases are key mediators of inflammation, highlighting the potential of kinase inhibitors as treatments for inflammatory disorders. Selective kinase inhibitors, however, have proved disappointing, particularly in the treatment of rheumatoid arthritis and inflammatory bowel disease. Consequently, to improve efficacy, attention has turned to multikinase inhibition.. The activity of a narrow spectrum kinase inhibitor, TOP1210, has been compared with selective kinase inhibitors (BIRB-796, dasatinib and BAY-61-3606) in a range of kinase assays, inflammatory cell assays, and in inflamed biopsies from patients with ulcerative colitis (UC). Effects on recombinant P38α, Src, and Syk kinase activities were assessed using Z-lyte assays (Invitrogen, Paisley, United Kingdom). Anti-inflammatory effects were assessed by measurement of proinflammatory cytokine release from peripheral blood mononuclear cells, primary macrophages, HT29 cells, inflamed colonic UC biopsies, and myofibroblasts isolated from inflamed colonic UC mucosa.. TOP1210 potently inhibits P38α, Src, and Syk kinase activities. Similarly, TOP1210 demonstrates potent inhibitory activity against proinflammatory cytokine release in each of the cellular assays and the inflamed colonic UC biopsies and myofibroblasts isolated from inflamed colonic UC mucosa. Generally, the selective kinase inhibitors showed limited and weaker activity in the cellular assays compared with the broad inhibitory profile of TOP1210. However, combination of the selective inhibitors led to improved efficacy and potency in both cellular and UC biopsy assays.. Targeted, multikinase inhibition with TOP1210 leads to a broad efficacy profile in both the innate and adaptive immune responses, with significant advantages over existing selective kinase approaches, and potentially offers a much improved therapeutic benefit in inflammatory bowel disease.

Topics: Benzamides; Biopsy; Colitis, Ulcerative; Cytokines; Dasatinib; HT29 Cells; Humans; Interleukin-8; Leukocytes, Mononuclear; Macrophages; Mitogen-Activated Protein Kinase 14; Myofibroblasts; Naphthalenes; Niacinamide; Primary Cell Culture; Protein Kinase Inhibitors; Pyrazoles; Pyrimidines; src-Family Kinases; Syk Kinase

Triggering receptor expressed on myeloid cells 1 (TREM-1) is a potent amplifier of inflammation. Recently, the antimicrobial peptide PGLYRP-1 was shown to be the ligand of TREM-1. Here, the ability of an anti-TREM-1 antibody to dampen the release of proinflammatory cytokines by colon lamina propria cells (LPCs) from patients with IBD was investigated and correlated with PGLYRP-1 levels.. Biopsies from patients with ulcerative colitis (UC, n = 45) or Crohn's disease (CD, n = 26) were compared with those from individuals undergoing colonoscopy for other reasons (n = 17). TREM-1 expression was analyzed on myeloid cells by flow cytometry. Cell culture experiments with LPCs were used to analyze PGLYRP-1 and inflammatory cytokine levels and assess the effect of anti-TREM-1 on cytokine secretion.. The frequency of TREM-1-expressing neutrophils and recruited macrophages was higher in inflamed than in noninflamed biopsies. The PGLYRP-1 level in inflamed tissue was higher than in noninflamed tissue; it was produced primarily by neutrophils, and its level correlated with the secretion of proinflammatory cytokines. Secretion of myeloperoxidase, tumor necrosis factor-α, interleukin-1β, and interleukin-8 by LPCs stimulated with the potent TREM-1 agonist consisting of PGLYRP-1 complexed with peptidoglycan was reduced in the presence of anti-TREM-1. Moreover, a blocking effect of anti-TREM-1 was apparent when LPCs from a subset of inflamed individuals with elevated PGLYRP-1 were stimulated with killed bacteria.. An anti-TREM-1 antibody can dampen secretion of proinflammatory cytokines in inflamed patients with elevated PGLYRP-1. Moreover, PGLYRP-1 + myeloperoxidase is a potential biomarker for predicting the effect of anti-TREM-1 therapy.

Topics: Adult; Aged; Antibodies; Biomarkers; Case-Control Studies; Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Cytokines; Female; Humans; Interleukin-1beta; Interleukin-8; Intestinal Mucosa; Macrophages; Male; Middle Aged; Neutrophils; Peptidoglycan; Peroxidase; Triggering Receptor Expressed on Myeloid Cells-1; Tumor Necrosis Factor-alpha; Young Adult

Vitamin D exerts anti-inflammatory actions both in vitro and in murine models of colitis. In previous studies, we demonstrated that vitamin D protects against the development of colitis by maintaining the integrity of the intestinal mucosal barrier.. We sought to evaluate whether deficient serum 25 hydroxyvitamin D [25(OH)D] concentrations are associated with increased mucosal inflammation, a loss of epithelial junctional proteins, and an increase in mucosal inflammatory cytokines in patients with ulcerative colitis (UC).. We prospectively enrolled 230 subjects with UC. Serum 25(OH)D concentrations were compared with the Mayo endoscopic score, the total Mayo score, and histologic activity. Colonic mucosal expression concentrations of vitamin D receptor (VDR), E-cadherin, zonula occluden 1 (ZO-1), occludin, claudin-2, tumor necrosis factor α (TNF-α), and interleukin 8 (IL-8) were compared between dichotomous groups with low or high serum 25(OH)D concentrations.. The mean serum 25(OH)D concentration was 21.8 ng/mL. Subjects stratified by concentrations included 12.6% ≥30 ng/mL, 45.6% ≥20 to <30 ng/mL, 37.4% ≥10 to <20 ng/mL, and 4.4% <10 ng/mL. There was an inverse association between serum 25(OH)D concentrations and mucosal inflammation as assessed by the Mayo endoscopy score (P = 0.01), disease activity as indicated by the total Mayo score (P = 0.001), and histologic activity (P = 0.02). A serum 25(OH)D concentration <20 ng/mL was associated with decreased mucosal transcript and protein expression concentrations of VDR, E-cadherin, and occludin as well as decreased protein expression of ZO-1, whereas TNF-α and IL-8 mucosal transcript expression concentrations were increased.. In UC patients, serum 25(OH)D concentration is inversely correlated with mucosal inflammation and disease activity. These results, coupled with the findings that serum 25(OH)D concentrations correlate with the mucosal expression of VDR as well as epithelial junction proteins and inversely with proinflammatory cytokines, suggest that vitamin D deficiency may contribute to UC inflammation by disrupting epithelial barrier function.

Topics: Adult; Antigens, CD; Cadherins; Colitis, Ulcerative; Colon; Female; Humans; Inflammation; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Occludin; Receptors, Calcitriol; Tumor Necrosis Factor-alpha; Vitamin D; Vitamin D Deficiency; Zonula Occludens-1 Protein

To explore Chinese medical theory of Fei and Dachang being interior-exteriorly correlated by observing changes of inherent immune response and acquired immune response in the lung tissue and the intestinal tissue of ulcerative colitis (UC) model rats and the intervention of Chinese compounds (CM).. Seventy rats were randomly divided into 5 groups, i.e., the normal control group (n = 10), the model group (n = 15), the treatment 1 group (n = 15, treated from Fei), the treatment 2 group (n = 15, treated from the intestine), and the Western medicine (WM) group [n = 15, treated with Sulfasalazine (SASP). Except those in the normal control group, the UC rat model was prepared by allergizing colon mucosa combined with TNBS-alcohol (50%) enema, and then intervened by medication (treated with CM complex prescription of treatment from lung, CM complex prescription of treatment from intestine, and SASP). After intragastric administration for 4 weeks, rats were sacrificed and samples taken. The expression of tumor necrosis factor α (TNF-α) and IL-8 contents in the lung tissue, the intestinal tissue, and the serum were detected by radioimmunoassay. Serum MedCAM-1 contents were detected using ELISA. Changes of the expression of Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB), neutrophil migration inhibition factor (MIF), mucosal addressin cell adhesion molecule-1 (MadCAM-1) mRNA in the lung tissue and the intestinal tissue were detected by real time PCR.. Compared with the normal control group, the expression levels of TNF-α, TLR4 mRNA, IL-8, MIF mR- NA, and MadCAM-1 mRNA obviously increased in the model group (P < 0.01). Compared with the model group, the expression levels of TNF-α, TLR4 mRNA, IL-8, MIF mRNA, and MadCAM-1 mRNA obviously decreased in the treatment 1 and 2 groups (P < 0.01). The expression of MadCAM-1 mRNA in the intestinal tissue was obviously higher in the model group than in the normal control group (P < 0.01), while the expressions of TNF-α and NF-κB mRNA was obviously lower in the model group than in the normal control group (P < 0.05, P < 0.01). Compared with the model group, the expression of MadCAM-1 mRNA all significantly deceased in each treatment group (P < 0.05, P < 0.01). Serum TNF-α contents were higher in the model group than in the normal control group (P < 0.05). Compared with the model group, serum TNF-α contents could be lowered in the treatment 1 and 2 groups (P < 0.05, P < 0.01).. The main mechanisms of the intestinal injury in this UC model might be related with activation of acquired immune response, accompanied with lowered functions of inherent immune response. The main mechanisms of the lung injury in this UC model might be related acquired immune response and inherent immune response. Treatment from Fei and treatment from Dachang both could obviously improve the immunodissonance of Fei and Dachang, indicating the special relation between the lung tissue and the intestinal tissue, thus providing experimental evidence for Chinese medical theory of Fei and Dachang being interior-exteriorly correlated.

Topics: Aleurites; Animals; Colitis, Ulcerative; Drugs, Chinese Herbal; Enema; Interleukin-8; Intestinal Mucosa; Intestines; Lung; Lung Injury; NF-kappa B; Rats; RNA, Messenger; Tumor Necrosis Factor-alpha

Ileal lesions of Crohn's disease [CD] patients are colonised by adherent-invasive Escherichia coli [AIEC] able to survive in macrophage cell lines. We analysed the ability of monocyte-derived macrophages [MDM] from CD patients to control AIEC intracellular replication and the pro-inflammatory cytokine response of the infected-MDM.. Peripheral blood MDM were obtained from 24 CD genotyped for NOD2 and ATG16L1 mutations, 5 ulcerative colitis [UC] patients and 12 healthy controls [HC]. The numbers of intracellular bacteria were determined using gentamicin assay. Cytokine secretion was quantified by ELISA assay.. We observed that higher levels of bacteria were internalised within MDM from CD patients than MDM from HC or UC patients. MDM from CD patients were unable to restrict AIEC intracellular replication. Infection of MDM from CD patients with AIEC resulted in significantly increased secretion of IL-6 and tumour necrosis factor alpha [TNF α] than did infection with non-pathogenic E. coli. AIEC-infected MDM from CD patients exhibited a disordered cytokines secretion compared with MDM from UC patients and HC. AIEC-infected MDM from patients with quiescent CD released significantly higher amounts of IL-6 and TNF-alpha than those with active disease or those from HC. The level of secreted TNF-alpha was correlated to the number of intracellular AIEC in MDM from CD patients. Treatment of MDM with infliximab did not change the MDM behaviour.. MDM from CD patients are unable to restrict intracellular AIEC replication, leading to disordered inflammatory response influenced by disease activity.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Autophagy-Related Proteins; Bacterial Adhesion; Bacterial Load; Carrier Proteins; Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Escherichia coli; Escherichia coli Infections; Female; Genotype; Humans; Infliximab; Interleukin-6; Interleukin-8; Macrophages; Male; Middle Aged; Nod2 Signaling Adaptor Protein; Polymorphism, Single Nucleotide; Tumor Necrosis Factor-alpha

Topics: Child; Colitis, Ulcerative; Colonoscopy; Drug Resistance; Female; Glucocorticoids; Humans; Interleukin-6; Interleukin-8; Leukapheresis; Protein Array Analysis; Treatment Outcome; Tumor Necrosis Factor-alpha

Inflammatory bowel diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC), are characterized by chronic intestinal inflammation due to immunological, microbial, and environmental factors in genetically predisposed individuals. Advances in the diagnosis, prognosis, and treatment of IBD require the identification of robust biomarkers that can be used for molecular classification of diverse disease presentations. We previously identified five genes, RELA, TNFAIP3 (A20), PIGR, TNF, and IL8, whose mRNA levels in colonic mucosal biopsies could be used in a multivariate analysis to classify patients with CD based on disease behavior and responses to therapy.. We compared expression of these five biomarkers in IBD patients classified as having CD or UC, and in healthy controls.. Patients with CD were characterized as having decreased median expression of TNFAIP3, PIGR, and TNF in non-inflamed colonic mucosa as compared to healthy controls. By contrast, UC patients exhibited decreased expression of PIGR and elevated expression of IL8 in colonic mucosa compared to healthy controls. A multivariate analysis combining mRNA levels for all five genes resulted in segregation of individuals based on disease presentation (CD vs. UC) as well as severity, i.e., patients in remission versus those with acute colitis at the time of biopsy.. We propose that this approach could be used as a model for molecular classification of IBD patients, which could further be enhanced by the inclusion of additional genes that are identified by functional studies, global gene expression analyses, and genome-wide association studies.

Topics: Adolescent; Adult; Aged; Chi-Square Distribution; Colitis, Ulcerative; Crohn Disease; DNA-Binding Proteins; Female; Gene Expression; Genetic Markers; Humans; Interleukin-8; Intestinal Mucosa; Intracellular Signaling Peptides and Proteins; Male; Middle Aged; Multivariate Analysis; Nuclear Proteins; Phenotype; Receptors, Polymeric Immunoglobulin; RNA, Messenger; Transcription Factor RelA; Tumor Necrosis Factor alpha-Induced Protein 3; Tumor Necrosis Factor-alpha; Young Adult

Infliximab is an established therapy for ulcerative colitis (UC). The aim of this study was to examine various serum cytokine levels and to identify possible markers predictive of therapeutic efficacy of infliximab for UC patients.. Twenty-one patients with moderately active UC were given intravenous infliximab (5 mg/kg) at 0, 2, and 6 weeks as induction therapy. The serum levels of 17 cytokines were determined using a Bio-Plex suspension array system before and 8 weeks after induction therapy. Partial Mayo score (PMS) and serum C-reactive protein levels were used for the determination of clinical activities at 0 and 8 weeks after the treatment. The overall therapeutic effect was determined at 26 weeks according to the PMS.. The median value of the PMS decreased significantly 8 weeks after the treatment (from 6 to 1.5, P < 0.05). However, C-reactive protein levels did not change significantly. Levels of serum interleukin (IL)-8 (P < 0.05) and macrophage inflammatory protein-1β (P < 0.005) significantly decreased 8 weeks after the induction. Serum levels of the other 15 cytokines did not change significantly. At 26 weeks, 13 of 20 patients (65%) were responders while 7 patients were non-responders. Levels of serum IL-6 at 8 weeks were significantly lower in responders than in non-responders (P < 0.05).. Serum IL-8 and macrophage inflammatory protein-1β seem to be sensitive markers for UC patients treated with infliximab, while IL-6 at 8 weeks after induction therapy may be predictive of subsequent response to infliximab.

Topics: Biomarkers; C-Reactive Protein; Chemokine CCL4; Colitis, Ulcerative; Cytokines; Forecasting; Gastrointestinal Agents; Induction Chemotherapy; Infliximab; Interleukin-6; Interleukin-8; Time Factors

Although current medications for ulcerative colitis (UC) are effective to some extent, there are still some limitation of their use due to the non-specific distribution, drug metabolism in the gastrointestinal tract, and severe adverse effects. In our previous studies, we developed oral redox nanoparticles (RNP(O)) that specifically accumulated and scavenged overproduced reactive oxygen species (ROS) in an inflamed colon. However, the mechanism leading to specific accumulation of RNP(O) in an inflamed colon is still unclear. In this study, we investigated the cellular uptake of RNP(O) into ROS-treated epithelial colonic cells in vitro, and compared to the untreated cells, found a significantly increased uptake in ROS-treated cells. In vivo, we discovered that orally administered RNP(O) were not internalized into the cells of a normal colon. A significant amount of disintegrated RNP(O) was detected in the cells of an inflamed colon of dextran sodium sulfate (DSS)-induced colitis mice, resulting in scavenging of ROS and suppression of inflammation with low adverse effects. Furthermore, we confirmed a significant reduction of disease activity and a robust dose response efficacy following RNP(O) treatment in acute DSS-induced colitis mice, outperforming the positive control 5-aminosalicylic acid. Oral administration of RNP(O) is a promising approach to develop a new therapy for UC disease.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Caco-2 Cells; Colitis, Ulcerative; Colon; Dextran Sulfate; Dose-Response Relationship, Drug; Humans; Hydrogen Peroxide; Interleukin-8; Leukocyte L1 Antigen Complex; Male; Mice; Mice, Inbred ICR; Nanoparticles; Oxidants; Oxidation-Reduction

Ulcerative colitis (UC) is associated with differential expression of genes involved in inflammation and tissue remodeling, including FOXO3a, which encodes a transcription factor known to promote inflammation in several tissues. However, FOXO3a expression in tissues affected by UC has not been examined. This study investigated the effects of FOXO3a on UC pathogenesis.. FOXO3a expression, in 23 patients with UC and in HT29 cells treated with tumor necrosis factor-α (TNF-α) for various durations, was detected by quantitative real-time polymerase chain reaction and Western blotting analysis. Enzyme-linked immunosorbent assay was used to quantify interleukin (IL)-8 expression in FOXO3a-silenced HT29 cells treated with TNF-α for various durations.. The messenger RNA and protein expression of FOXO3a were significantly lower in UC tissues than those in normal subjects (P < 0.01). TNF-α treatment for 0, 0.5, 1, 6, and 24 h induced FOXO3 degradation in HT29 cells. FOXO3a silencing increased IL-8 levels in HT29 cells treated with TNF-α for 6 h (P < 0.05).. FOXO3a may play an important role in the intestinal inflammation of patients with UC.

Topics: Adult; Blotting, Western; Colitis, Ulcerative; Enzyme-Linked Immunosorbent Assay; Female; Forkhead Box Protein O3; HT29 Cells; Humans; Inflammation; Interleukin-8; Intestinal Mucosa; Intestines; Male; Middle Aged; Real-Time Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

Background. Patients with UC have shown an important defect in the secretion and maintenance of the mucosal barrier as part of inadequate expression of mucin genes. The aim of the present study was to determine the expression of MUC12, MUC16, and MUC20 in colonic tissue from patients with UC in regard to their clinical outcomes. Methods. We included a total of 40 patients with UC and 30 normal controls. Mucin gene expression was performed by RT-PCR and protein expression was detected by immunohistochemistry. Results. Patients with active UC showed no significant expression of MUC12 gene in mucosa compared to the group of patients with UC in remission and the normal control group. MUC16 gene expression was significantly increased in the UC active and remission groups compared to the normal control group (P = 0.03). MUC20 gene expression was found significantly decreased in patients with active UC compared to both remission group (P = 0.001) and normal controls (P = 0.001). Furthermore, an association was found between MUC20 gene expression and the presence of histological remission in patients with UC (P = 0.003, OR = 0.37). Conclusions. An increased gene expression of MUC16 and MUC20 was found in patients with remission UC.

Topics: Adult; Aged; CA-125 Antigen; Colitis, Ulcerative; Colon; Female; Gene Expression Regulation; Humans; Interleukin-6; Interleukin-8; Male; Membrane Proteins; Middle Aged; Mucins; RNA, Messenger

Ulcerative colitis (UC) is idiopathic, chronic and relapsing inflammatory bowel disease. Factors which initiate and perpetuate UC are not well understood. It is still unclear if any relationship exists between cytokines, oxidative stress, gastrointestinal (GI) motility, and small intestinal bacterial overgrowth (SIBO) in UC patients.. To examine the relationship between these factors among UC patients.. A total of 120 UC patients and 125 age and sex matched controls with no GI symptoms were enrolled. Plasma levels of IL-6, IL-8, TNF-α and IL-10 were measured in all subjects by using ELISA. Lipid peroxidation (LPO) and reduced glutathione (GSH) were measured by standard methods. Orocecal transit time (OCTT) and SIBO were measured by lactulose and glucose hydrogen breath tests respectively.. Out of the 120 UC patients, 74 were male with mean±SD age of 45.6±17.5years. Plasma levels of IL-6, IL-8, TNF-α and IL-10 in UC patients were significantly higher (p<0.01) as compared to controls. LPO in UC patients was significantly increased (p<0.01) while GSH was significantly decreased (p<0.01) as compared to controls. OCTT and SIBO were significantly higher in UC patients as compared to controls. UC patients with elevated inflammatory cytokines showed delayed OCTT and increased SIBO. It was also observed that there was a significant correlation between SIBO with IL-6, IL-8, TNF-α, and IL-10, LPO and GSH.. This study indicates that increase in cytokines and decrease in anti-oxidants in UC patients would have resulted in oxidative stress causing delayed GI motility leading to SIBO.

Topics: Adult; Aged; Case-Control Studies; Colitis, Ulcerative; Cytokines; Enzyme-Linked Immunosorbent Assay; Gastrointestinal Motility; Gastrointestinal Transit; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Lipid Peroxidation; Middle Aged; Oxidative Stress; Tumor Necrosis Factor-alpha; Young Adult

MicroRNAs (miRNAs) are important posttranscriptional regulators of gene expression. The precise role of miRNAs in ulcerative colitis (UC) is not completely understood. The purpose of this study was to identify miRNAs that are induced in patients with active UC and to assess the effect of miR-155 on improving intestinal inflammation.. The miRNA profiles in patients with active UC (n = 20) and healthy subjects (n = 16) were examined using miRNA microarrays. miR-155 upregulation was confirmed by quantitative RT-PCR. Regulation of the target gene FOXO3a expression by miR-155 was assessed using luciferase reporter construct assays and miR-155 mimic or inhibitor transfections. The effects of FOXO3a or miR-155 on IκBα or IL-8 were detected by Western blot or enzyme-linked immunosorbent assay in HT29 cells, respectively.. We identified 68 miRNAs that were differentially expressed (33 upregulated and 35 downregulated) in patients with active UC compared with healthy controls. One of the upregulated miRNAs in the UC tissue was miR-155 (1.22-fold, P < 0.03), which plays a key role in the regulation of inflammatory pathways. In patients with active UC, miR-155 was significantly upregulated, and the expression of FOXO3a dramatically decreased. Luciferase reporter assays demonstrated that miR-155 directly targets FOXO3a and affects the protein expression of FOXO3a in HT29 cells. Moreover, silenced FOXO3a and the overexpression of miR-155 increased the levels of IL-8 in TNF-α-treated HT29 cells by suppressing the inhibitory IκBα.. miR-155 appears to play a role in the intestinal inflammation of patients with active UC by downregulating the expression of FOXO3a. This process may activate the nuclear factor kappa B signaling pathway.

Topics: 3' Untranslated Regions; Adult; Case-Control Studies; Colitis, Ulcerative; Down-Regulation; Female; Forkhead Box Protein O3; Forkhead Transcription Factors; Gene Expression Profiling; Gene Expression Regulation; Gene Knockdown Techniques; HT29 Cells; Humans; I-kappa B Proteins; Interleukin-8; Male; MicroRNAs; Middle Aged; NF-KappaB Inhibitor alpha; Oligonucleotide Array Sequence Analysis; Signal Transduction; Tumor Necrosis Factor-alpha; Up-Regulation

CXCL16 is a scavenger receptor which has been connected to phagocytosis of bacterial antigens in experimental colitis. It has also been shown to have a pivotal role in the development of experimental colitis in mice. The increased expression of CXCL16 has been demonstrated in inflamed lesions of patients with Crohn disease. Our aim was to study the expression of CXCL16 in the colon of patients with ulcerative colitis.. Relative quantitative reverse transcription-polymerase chain reaction was applied to explore the gene expressions of CXCL16, its receptor CXCR6, and interleukin 8, an inflammatory marker, in the colonic biopsies of children with active ulcerative colitis (n=19), children with ulcerative colitis in remission (n=9) and children with no inflammatory condition in colon (n=14).. An increased expression of CXCL16 in the colonic biopsies of children with ulcerative colitis was found both in active disease (p=0.006) and in remission (p=0.033), when compared to children without inflammatory condition. The gene expressions of interleukin 8 and CXCL16 correlated with each other (rs=0.67, p=0.01). The expression of CXCR6 mRNA was comparable between the study groups (p=0.50).. The gene expression of CXCL16 was increased in patients with ulcerative colitis both in active disease and in remission suggesting an important role of the molecule in the pathogenesis of the condition.

Topics: Adolescent; Chemokine CXCL16; Chemokines, CXC; Child; Child, Preschool; Colitis, Ulcerative; Gene Expression; Humans; Interleukin-8; Receptors, Chemokine; Receptors, CXCR6; Receptors, Scavenger; Receptors, Virus; RNA, Messenger

[. To observe the effect of moxibustion at different duration on colonic mucosal morphological chan-ObjectiveTo observe the effect of moxibustion at different duration on colonic mucosal morphological changes, serum and colonic cytokine levels in ulcerative colitis (UC) rats, so as to provide experimental evidence for clinical treat-ges, serum and colonic cytokine levels in ulcerative colitis (UC) rats, so as to provide experimental evidence for clinical treatment of UC.. SD rats were randomly divided into blank control, UC model, 3 cones-moxibustion (3-cones-M), 6-SD rats were randomly divided into blank control, UC model, 3 cones-moxibustion (3-cones-M), 6-cones-M and 9-cones-M groups, with 6 rats in each group. UC model was established by intra-rectal injection of mixture solution ofcones-M and 9-cones-M groups, with 6 rats in each group. UC model was established by intra-rectal injection of mixture solution of 5% trinitro-benzene-sulfonic acid (TNBS, 100 mg/kg) and 50% alcohol (1 1) under anesthesia and oral administration of 5%5% trinitro-benzene-sulfonic acid (TNBS, 100 mg/kg) and 50% alcohol (1 : 1) under anesthesia and oral administration of 5% dextran sodium sulfate. Moxibustion (ignited moxa cones) was applied to "Tianshu" (ST 25) and "Daheng" (SP 15), once daily indextran sodium sulfate. Moxibustion (ignited moxa cones) was applied to "Tianshu" (ST 25) and "Daheng" (SP 15), once daily in the first 7 days, and once every other day in the subsequent 14 days. Serum IL-8 and IL- 10 contents were assayed by ELISA andthe first 7 days, and once every other day in the subsequent 14 days. Serum IL-8 and IL-10 contents were assayed by ELISA and colonic toll-like receptor 9 (TLR-9) and nuclear factor-icB p 65 (NE-KB p 65) protein expression levels detected by Western blot.colonic toll-like receptor 9 (TLR-9) and nuclear factor-mB p 65 (NF-mB p 65) protein expression levels detected by Western blot. The colonic mucosal structure was observed by light microscope after H. E. staining, and by electron microscope, respectively.The colonic mucosal structure was observed by light microscope after H. E. staining, and by electron microscope, respectively.. In comparison with the blank control group, the Disease Activity Index (DAI), serum IL-8 content, colonic TLR-9 andResults - In comparison with the blank control group, the Disease Activity Index (DAI), serum IL-8 content, colonic TLR-9 and NE-KB p 65 protein expression levels were significantly increased in the model group ( P<0. 05), and serum IL-la content wasNF-mB p 65 protein expression levels were significantly increased in the model group ( P < 0.05), and serum IL-10 content was notably decreased in the model group (P < 0.05). While in comparison with the model group, the DAI, serum IL-8 content, co-notably decreased in the model group (P<0.05). While in comparison with the model group, the DAI, serum IL-8 content, coIonic TLR-9 and NE-kappaB p 65 protein expression levels in the 3-cones-M, 6-cones-M and 9-cones-M groups were remarkably down-lonic TLR-9 and NF-mB p 65 protein expression levels in the 3-cones-M. 6-cones-M and 9-cones-M groups were remarkably down- regulated (P < 0.05), and serum IL-10 contents considerably up-regulated in the three moxibustion groups (P < 0.05). No significant differences were found among the three moxibustion groups in the DAI (P > 0.05). The serum IL-8 contents were significantly lower and serum IL-10 contents were considerably higher in the 6-cones-M and 9-cones-M groups than in the 3-cones-M group (P < 0.05). The changes of colonic TLR-9 and NF-kappaB p 65 protein expression were more remarkable in the 9-cones-M group than in the 3-cones-M and 6-cones-M groups (P < 0.05). Results of H.E. staining and electron microscopy showed that in the model group, mucosal injury, partial disorganization of the glandular organ, edema and congestion and inflammatory cell infiltration, mucosal epithelial microvili injury with disordered arrangement, etc. under light microscope, and local mucosal defect, apoptotic bodies and mucolysis under electron microscope were found, these situations were obviously lighter in rats of the three moxibustion groups, particularly in the 9-cones-M group.. Moxibustion intervention can relieve colonic mucosal injury in UC mice, which may be closely associated with its effects in suppressing serum proinflammatory cytokine IL-8, up-regulating anti-inflammatory cytokine IL-10 level, and inhibiting colonic NF- KB p 65 and TLR-9 protein expression, and the effects of longer duration of moxibustion are better.

Topics: Acupuncture Points; Animals; Colitis, Ulcerative; Colon; Cytokines; Disease Models, Animal; Female; Humans; Interleukin-10; Interleukin-8; Intestinal Mucosa; Male; Moxibustion; Rats; Rats, Sprague-Dawley; Signal Transduction; Toll-Like Receptor 9

To investigate the efficacy of moxibustion in ulcerative colitis (UC) rats from morphological, immunological and molecular biological perspectives.. Thirty-two Sprague-Dawley rats were randomly assigned to a blank control group (normal rats, n = 6) and a model replication (MR) group (UC rats, n = 26). A UC model was established by 2,4,6-trinitrobenzenesulfonic acid/dextran sulfate sodium enema. Rats in the MR group were further randomly assigned to a 9-min moxibustion (9M) group (9 moxa-cone, n = 6), 6-min moxibustion (6M) group (6 moxa-cone, n = 6), 3-min moxibustion (3M) group (3 moxa-cone, n = 6), and a waiting list control (WLC) group (no moxibustion treatment, n = 6). Rats in the moxibustion treatment group were treated in 14 sessions over 28 d. Disease activity, local tissue morphology, serum level of interleukin (IL)-8 and IL-10, and expression of Toll-like receptor (TLR)9 as well as nuclear factor (NF)-κB p65 in colonic tissue were determined by disease activity index (DAI), hematoxylin and eosin staining, electron microscopy, enzyme-linked immunosorbent assay and Western blotting, respectively.. DAI was lowest in the 9M group and highest in the WLC group. The differences in DAI between the moxibustion treatment (3M, 6M, 9M) and no treatment groups were significant for all one-to-one comparisons (0.60 ± 0.54 vs 1.20 ± 0.44, 0.60 ± 0.54 vs 1.80 ± 0.45, 0.60 ± 0.54 vs 3.0 ± 0.45, respectively, P < 0.05). Light and electron microscopy showed that the neatness of the glandular arrangement in colonic mucosal epithelia gradually increased in the WLC, 3M, 6M to 9M groups. IL-8 level successively decreased while IL-10 level increased from the WLC to 3M, 6M and 9M groups. The differences among these groups were significant for all comparisons (105.46 ± 8.75 vs 76.61 ± 3.58, 105.46 ± 8.75 vs 69.78 ± 1.87, 105.46 ± 8.75 vs 67.41 ± 1.84, respectively, P < 0.01 for IL-8; and 30.83 ± 1.29 vs 75.64 ± 1.90, 30.83 ± 1.29 vs 80.90 ± 3.16, 30.83 ± 1.29 vs 83.46 ± 2.37, respectively, P < 0.01 for IL-10), except comparison of 6M vs 9M. Expression of TLR9 and NF-κB p65 decreased in order: highest in the WLC group and lowest in the 9M group. In addition, the differences among the WLC, 3M, 6M and 9M groups were significant for all comparisons (0.492 ± 0.026 vs 0.380 ± 0.022, 0.492 ± 0.026 vs 0.355 ± 0.005, 0.492 ± 0.026 vs 0.327 ± 0.015, respectively, P < 0.05 for TLR9; and 0.436 ± 0.041 vs 0.326 ± 0.022, 0.436 ± 0.041 vs 0.293 ± 0.006, 0.436 ± 0.041 vs 0.265 ± 0.017, respectively, P < 0.05 for NF-κB p65).. Moxibustion repairs damaged colonic mucosa, suppresses serum IL-8, activates serum IL-10 level, and decreases expression of TLR-9 and NF-κB p65 in UC rats.

Topics: Animals; Colitis, Ulcerative; Colon; Dextran Sulfate; Disease Models, Animal; Female; Inflammation Mediators; Interleukin-10; Interleukin-8; Intestinal Mucosa; Male; Moxibustion; Rats, Sprague-Dawley; Time Factors; Toll-Like Receptor 9; Transcription Factor RelA; Trinitrobenzenesulfonic Acid; Wound Healing

To observe the effect of total flavonoids of Oldenlendia difflusa (FOD) on NF-kappaB and IL-8, TNF-alpha, IL-10 expressions of ulcerative colitis (UC) model rats, and explore its immunological mechanism of anti-UC.. Sixty Kunming male mice with the average weight of (20 +/- 2) g were randomly divided into six groups. The control group (cont) was orally administered with distilled water. Whereas the remaining five groups were fed with 4% dextran sulphate sodium (DSS) solution for seven days to induce acute UC, and orally administered with the following drugs: distilled water (for the DSS group), SASP at dose of 500 mg x kg(-1) x d(-1) for the DSS + SASP group, FOD at dose of 60 mg x kg(-1) x d(-1) for the DSS + FOD-H group, FOD at dose of 40 mg x kg(-1) x d(-1) for the DSS + FOD-M group, and FOD at dose of 26.7 mg x kg(-1) x d(-1) for the DSS + FOD-L group. During the modeling and drug administration, the mice were scored for DAI. Seven days later, the mice were put to death, and their colonic tissue samples were collected to evaluate colonic mucosal lesions. The NF-kappaB p65, IL-8, TNF-alpha, IL-10 expressions were tested by immunohistochemical staining and ELISA.. Seven-day feeding with 4% DSS solution could successfully induce acute UC in mice. Compared with the cont group, the DSS group showed significantly higher DAI and colonic mucosal lesions, remarkable increase in NF-kappaB p65, IL-8, TNF-alpha expression in colonic tissues, and notable decrease in IL-10 expression (P < 0.05). FOD could prevent acute UC in mice included by DSS. Seven-day administration of 60 mg x kg(-1) x d(-1) or 40 mg x kg(-1) x d(-1) FOD could completely or partially resist the above mentioned changes caused by DSS. Compared with the DSS group, the DSS + FOD-H group and the DSS + FOD-M group showed reduction in colonic mucosal lesions, down-regulation in IL-8, TNF-alpha and NF-kappaB p65 expressions and up-regulation in IL-10 expression (P < 0.05).. FOD could significantly resist UC in mice. Its mechanism may be related to the inhibition of NF-kappaB p65 activation, the reduction of IL-8 and TNF-alpha expressions and the increase in the anti-inflammatory factor IL-10.

Topics: Animals; Anti-Inflammatory Agents; Colitis, Ulcerative; Drugs, Chinese Herbal; Flavonoids; Humans; Interleukin-8; Male; Mice; NF-kappa B; Oldenlandia; Transcription Factor RelA; Tumor Necrosis Factor-alpha

We examined the relationship among the multidrug resistance (MDR1) gene product P-glycoprotein (P-gp), ulcerative colitis, and immune status under ulcerative colitis. MDR1 P-gp expression and interleukin-8 levels in ulcerative colitis were determined using immunohistochemistry and a double-antibody sandwich avidin-biotin complex-enzyme-linked immunosorbent assay, respectively. Nitric oxide content and nitric oxide synthase activity in the colonic mucosa were determined using a colorimetric method; CD4(+) and CD25(+) T cell subset percentages in the peripheral blood were determined by flow cytometry. The positive expression rate of P-gp in patients with ulcerative colitis (17.4%) was significantly lower than that in the control group (31.4%). The expression rate decreased to 10.1, 9.2, and 8.3% after 12, 18, and 24 months of treatment, respectively, which were significantly lower than the expression rate before treatment (17.4%). P-gp expression levels during the remission phase and active phase of ulcerative colitis were 15.2 and 17.1%, respectively, which were significantly lower than that in normal controls (31.4%). Compared with P-gp-negative patients, nitric oxide content, nitric oxide synthase activity, and interleukin-8 levels were significantly higher in P-gp-positive patients with moderately active, severely active, early onset, chronic relapsing, chronic persistent, and acute fulminant ulcerative colitis. CD4(+) and CD25(+) T cell subsets were significantly lower in the peripheral blood of patients with severely active and acute fulminant ulcerative colitis than in control subjects. Expression of the multidrug resistance gene and its product P-gp was observed in normal colon tissues and may be closely related to ulcerative colitis.

Topics: Adolescent; Adult; Aged; ATP Binding Cassette Transporter, Subfamily B; Case-Control Studies; China; Colitis, Ulcerative; Female; Gastrointestinal Agents; Humans; Immunosuppressive Agents; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Nitric Oxide; Nitric Oxide Synthase; T-Lymphocyte Subsets; Young Adult

Ulcerative colitis (UC) is associated with differential colonic expression of genes involved in immune response (e.g. IL8) and barrier integrity (e.g. cadherins). MicroRNAs (miRNAs) are regulators of gene expression and are involved in various immune-related diseases. In this study, we investigated (1) if miRNA expression in UC mucosa is altered and (2) if any of these changes correlate with mucosal mRNA expression. Integration of mRNA and miRNA expression profiling may allow the identification of functional links between dysregulated miRNAs and their target mRNA.. Colonic mucosal biopsies were obtained from 17 UC (10 active and 7 inactive) patients and 10 normal controls. Total RNA was used to analyze miRNA and mRNA expression via Affymetrix miRNA 2.0 and Affymetrix Human Gene 1.0ST arrays, respectively. Both miRNA and gene expression profiles were integrated by correlation analysis to identify dysregulated miRNAs with their corresponding predicted target mRNA. Microarray data were validated with qRT-PCR. Regulation of IL8 and CDH11 expression by hsa-miR-200c-3p was determined by luciferase reporter assays.. When comparing active UC patients vs. controls, 51 miRNAs and 1543 gene probe sets gave significantly different signals. In contrast, in inactive UC vs. controls, no significant miRNA expression differences were found while 155 gene probe sets had significantly different signals. We then identified potential target genes of the significantly dysregulated miRNAs and genes in active UC vs. controls and found a highly significant inverse correlation between hsa-miR-200c-3p and IL8, an inflammatory marker, and between hsa-miR-200c-3p and CDH11, a gene related to intestinal epithelial barrier function. We could demonstrate that hsa-miR-200c-3p directly regulates IL8 and CDH11 expression.. Differential expression of immune- and barrier-related genes in inflamed UC mucosa may be influenced by altered expression of miRNAs. Integrated analysis of miRNA and mRNA expression profiles revealed hsa-miR-200c-3p for use of miRNA mimics as therapeutics.

Topics: Base Sequence; Biopsy; Cadherins; Case-Control Studies; Colitis, Ulcerative; Colon; Epithelial Cells; Female; Gene Expression Profiling; Gene Expression Regulation; Genes, Reporter; HT29 Cells; Humans; Inflammation; Interleukin-8; Intestinal Mucosa; Luciferases; Male; MicroRNAs; Middle Aged; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis; Real-Time Polymerase Chain Reaction; Reproducibility of Results; RNA, Messenger

To explore the efficacy of muscovite on iodoacetamide -induced ulcerative colitis in rats and elucidate its possible mechanism.. Ulcerative colitis was induced in female Sprague-Dawley (SD) rats by an intracolonic injection of iodoacetamide. A total of 48 rats were divided randomly(by the method of random digits table) into 6 groups: control group, model group, low-dose muscovite group (360 mg/kg), high-dose muscovite group (720 mg/kg), 5-aminosalicylie acid (5-ASA) group and muscovite plus 5-ASA group (combined treatment), and each group had 8 rats. The body weight, disease activity index (DAI), macroscopic damage and microscopic score of rats in each group were subsequently evaluated after dosing for 7 days. The protein levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8) and myeloperoxidase (MPO) activity were detected by enzyme-linked immunosorbent assay(ELISA) while the activity of nuclear facor(NF)-κB was determined by immunohistochemistry.One way ANOVA and rank-sum test were used.. After doing, body weight macroscopic damage, microscopic score, TNF-α concentration, MPO and NF-κB activity of rats in each group were all significantly correlated with the dose of muscovite (r = 0.573, -0.647, -0.569, -0.681, -0.811, -0.842, all P < 0.05). High-dose muscovite group had no significant difference with 5-ASA group in body weightt, DAI, macroscopic damage, microscopic score, IL-8 concentration, TNF-α concentration, MPO and NF-κB activities((166 ± 5) vs (167 ± 5) g, 0.33 (0.00, 1.17) vs 0.17 (0.00, 0.83), 2.50 (2.00, 4.00) vs 3.00 (2.00, 3.00), 3.00 (2.00, 3.00) vs 2.50 (2.00, 3.00), (109 ± 17) vs (111 ± 15) pg/ml, (166 ± 38) vs (155 ± 45) pg/ml, (52 ± 6) vs (49 ± 4) U/g, 7.39 ± 0.42 vs 7.41 ± 0.34, all P > 0.05). The MPO and NF-κB activities of combined treatment group were lower than those of 5-ASA group((40 ± 4) vs (49 ± 4) U/g, 4.67 ± 0.72 vs 7.41 ± 0.34, all P < 0.05). However, other indices showed no significant difference with 5-ASA group (all P > 0.05).. Rectal administration of muscovite ameliorates colonic inflammation of iodoacetamide-induced colitis. Its underlying mechanism is probably due to the regulation of inflammatory response. Muscovite may be a potential therapeutic agent for treatment of ulcerative colitis.

Topics: Aluminum Silicates; Animals; Colitis, Ulcerative; Enzyme-Linked Immunosorbent Assay; Female; Interleukin-8; NF-kappa B; Peroxidase; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha

To observe the effect of Qingchang Huashi Recipe (QHR) on the activation and expressions of nuclear factor kappaB (NF-kappaB), Toll-like receptors (TLRs), and contents of interleukin-8 (IL-8), thus exploring its possible mechanisms for treating ulcerative colitis (UC).. The HT-29 cells were induced to inflammation model by tumor necrosis factor-alpha (TNF-alpha) and lipopolysaccharides (LPS). HT-29 cells were divided into 6 groups, i.e., the vehicle control group, the model control group, the sulfasalazine (SASP) group, the high dose QHR group, the middle dose QHR group, the low dose QHR group. Effects on the cell growth were detected by MTT. The chemoattractant of macrophages was observed using Transwell. The expressions of NF-kappaB and TLR4 protein were detected using immune cell fluorescence method. The content of IL-8 was detected by ELISA.. The growth of cells were not inhibited in each group. Statistical difference existed in each dose QHR group in inhibiting the chemoattractant of macrophages, reducing activation of NF-kappaB, lowing expressions of TLR4 protein, and decreasing the secretion of IL-8, when compared with the model control group (P < 0.05). No statistical difference existed in inhibiting the chemoattractant of macrophages between the high dose QHR group and the vehicle control group (P > 0.05). But its inhibition on NF-kappaB activation was higher in the high dose QHR group than in the SASP group (P < 0.05).. QHR could obviously attenuate the inflammatory reaction of HT-29 cells, inhibit the chemoattractant of macrophages, reduce the activation of NF-kappaB, lower expressions of TLR-4, and attenuate the secretion of IL-8, which might be one of its mechanisms for treating UC.

Topics: Colitis, Ulcerative; Drugs, Chinese Herbal; HT29 Cells; Humans; Inflammation; Interleukin-8; NF-kappa B; Signal Transduction; Toll-Like Receptor 4

Topics: Adult; Biomarkers; Case-Control Studies; Colitis, Ulcerative; Colon; Humans; Interleukin-8; Intestinal Mucosa; Real-Time Polymerase Chain Reaction; RNA, Messenger; Up-Regulation

The aim of this study is to understand intracellular regulatory mechanisms in peripheral blood mononuclear cells (PBMCs), which are either common to many autoimmune diseases or specific to some of them. We incorporated large-scale data such as protein-protein interactions, gene expression and demographical information of hundreds of patients and healthy subjects, related to six autoimmune diseases with available large-scale gene expression measurements: multiple sclerosis (MS), systemic lupus erythematosus (SLE), juvenile rheumatoid arthritis (JRA), Crohn's disease (CD), ulcerative colitis (UC) and type 1 diabetes (T1D). These data were analyzed concurrently by statistical and systems biology approaches tailored for this purpose. We found that chemokines such as CXCL1-3, 5, 6 and the interleukin (IL) IL8 tend to be differentially expressed in PBMCs of patients with the analyzed autoimmune diseases. In addition, the anti-apoptotic gene BCL3, interferon-γ (IFNG), and the vitamin D receptor (VDR) gene physically interact with significantly many genes that tend to be differentially expressed in PBMCs of patients with the analyzed autoimmune diseases. In general, similar cellular processes tend to be differentially expressed in PBMC in the analyzed autoimmune diseases. Specifically, the cellular processes related to cell proliferation (for example, epidermal growth factor, platelet-derived growth factor, nuclear factor-κB, Wnt/β-catenin signaling, stress-activated protein kinase c-Jun NH2-terminal kinase), inflammatory response (for example, interleukins IL2 and IL6, the cytokine granulocyte-macrophage colony-stimulating factor and the B-cell receptor), general signaling cascades (for example, mitogen-activated protein kinase, extracellular signal-regulated kinase, p38 and TRK) and apoptosis are activated in most of the analyzed autoimmune diseases. However, our results suggest that in each of the analyzed diseases, apoptosis and chemotaxis are activated via different subsignaling pathways. Analyses of the expression levels of dozens of genes and the protein-protein interactions among them demonstrated that CD and UC have relatively similar gene expression signatures, whereas the gene expression signatures of T1D and JRA relatively differ from the signatures of the other autoimmune diseases. These diseases are the only ones activated via the Fcɛ pathway. The relevant genes and pathways reported in this study are discussed at length, and may be helpful in the d

Topics: Apoptosis; Arthritis, Juvenile; Autoimmune Diseases; B-Cell Lymphoma 3 Protein; Cell Proliferation; Chemokine CXCL1; Chemokine CXCL5; Chemokine CXCL6; Chemokines, CXC; Colitis, Ulcerative; Crohn Disease; Diabetes Mellitus, Type 1; Gene Expression; Humans; Inflammation; Interferon-gamma; Interleukin-8; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; MAP Kinase Signaling System; Mitogen-Activated Protein Kinases; Multiple Sclerosis; Protein Interaction Maps; Proto-Oncogene Proteins; Receptors, Calcitriol; Receptors, IgE; Signal Transduction; Transcription Factors; Transcriptome

Ulcerative colitis (UC) is associated with colorectal cancer. Chronic inflammation may also play a role in the pathogenesis of sporadic colorectal cancer (SCC), particularly in younger patients (<55 years). We evaluated whether single nucleotide polymorphisms of the OCTN1 and OCTN2 genes are associated with UC, SCC, and with UC cases with cancer progression (UCCP).. We evaluated the OCTN1 and OCTN2 polymorphisms in 200 patients with UC, 59 patients with UCCP, 200 patients with SCC, and 200 controls (HC). IL-8 expression was also assessed by real-time polymerase chain reaction (PCR). Additionally, we transfected human colon carcinoma Caco2 cells, homozygous for OCTN1/1672T variant, with the OCTN1/1672C allele and NF-κB activity was evaluated by luciferase based reporter assay and IL-8 mRNA expression by real-time PCR.. OCTN2 polymorphisms did not present a significant association with any group of patients compared to normal controls. Conversely, homozygosity for the OCTN1/1672T variant was significantly associated with UC (P = 0.047 vs. HC), with UCCP (UCCP vs. HC, P < 0.001), and with SCC developing in early age (<55 years) (P = 0.021 vs. HC). Importantly, IL-8 mRNA expression was higher in UC and UCCP patients homozygous for the OCTN1 1672T variant compared to the other genotypes. Moreover, in Caco2 cells transfection of the OCTN1/1672C variant reduced the activity of the proinflammatory factor NF-κB.. Our data demonstrate that OCTN1 could have a role in modulating the severity of chronic inflammation associated with SCC in early age and in UC patients, and that its polymorphisms may help to predict malignant progression of IBD.

Topics: Caco-2 Cells; Cell Transformation, Neoplastic; Chi-Square Distribution; Colitis, Ulcerative; Colorectal Neoplasms; Disease Progression; Female; Humans; Interleukin-8; Male; Middle Aged; NF-kappa B; Organic Cation Transport Proteins; Polymorphism, Single Nucleotide; RNA, Messenger; Solute Carrier Family 22 Member 5; Statistics, Nonparametric; Symporters

Crohn's disease (CD) and ulcerative colitis (UC), known as inflammatory bowel diseases (IBD), are characterized by an abnormal immunological response to commensal bacteria colonizing intestinal lumen and mucosa. Among the latter, strains of adherent-invasive Escherichia coli (AIEC), capable of adhering to and invading epithelium, and to replicate in macrophages, have been described in CD adults. We aimed at identifying and characterizing AIEC strains in pediatric IBD.. In all, 24 CD children, 10 UC, and 23 controls were investigated. Mucosal biopsies, taken during colonoscopy, were analyzed for the presence of AIEC strains by an adhesive-invasive test. Protein expression of the specific AIEC receptor, the carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), was evaluated by western blot and immunohistochemistry, while tumor necrosis factor alpha (TNF-α) and interleukin (IL)-8 mRNA expression was detected by real-time polymerase chain reaction (PCR), after bacterial infection. Transmission electron microscopy and trans-epithelial electric resistance assays were performed on biopsies to assess bacteria-induced morphological and functional epithelial alterations.. Two bacterial strains, EC15 and EC10, were found to adhere and invade the Caco2 cell line, similar to the well-known AIEC strain LF82 (positive control): they upregulated CEACAM6, TNF-α, and IL-8 gene/protein expression, in vitro and in cultured intestinal mucosa; they could also survive inside macrophages and damage the epithelial barrier integrity. Lesions in the inflamed tissues were associated with bacterial infection.. This is the first study showing the presence of adhesive-invasive bacteria strains in the inflamed tissues of children with IBD. Collective features of these strains indicate that they belong to the AIEC spectrum, suggesting their possible role in disease pathogenesis.

Topics: Adolescent; Animals; Antigens, CD; Bacterial Adhesion; Blotting, Western; Case-Control Studies; Cell Adhesion Molecules; Cells, Cultured; Child; Colitis, Ulcerative; Crohn Disease; Escherichia coli; Escherichia coli Infections; Female; Fluorescent Antibody Technique; GPI-Linked Proteins; Humans; Immunoenzyme Techniques; Interleukin-8; Intestinal Mucosa; Macrophages; Male; Mice; Organ Culture Techniques; Real-Time Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha

Milk fat globule-epidermal growth factor 8 (MFG-E8) plays an important role in maintaining intestinal barrier homeostasis and accelerating intestinal restitution. However, studies of MFG-E8 expression in humans with ulcerative colitis are lacking. We examined MFG-E8 expression in colonic mucosal biopsies from ulcerative colitis patients and healthy controls (n = 26 each) by real-time quantitative polymerase chain reaction (PCR), Western blot analysis and immunohistochemistry. MFG-E8 mRNA and protein expression was lower in ulcerative colitis patients than in controls. MFG-E8 expression was inversely correlated with mucosal inflammatory activity and clinical disease activity in patients. MFG-E8 was present in human intestinal epithelial cells both in vivo and in vitro. Apoptosis induction was also detected in the intestinal epithelium of ulcerative colitis patients by terminal-deoxynucleoitidyl transferase mediated nick-end labeling assay. We used lentiviral vectors encoding human MFG-E8 targeting short hairpin RNA to obtain MFG-E8 knockdown intestinal epithelia cell clones. MFG-E8 knockdown could promote apoptosis in intestinal epithelial cell lines, accompanied by a decrease in level of the antiapoptotic protein B-cell lymphoma 2 (BCL-2) and induction of the proapoptotic protein BCL2-associated protein X (BAX). The addition of recombinant human MFG-E8 led to decreased BAX and cleaved caspase-3 levels and induction of BCL-2 level in intestinal epithelia cells. MFG-E8 knockdown also attenuated wound healing on scratch assay of intestinal epithelial cells. The mRNA level of intestinal trefoid factor 3, a pivotal factor in intestinal epithelial cell migration and restitution, was downregulated with MFG-E8 knockdown. In conclusion, we demonstrated that decreased colonic MFG-E8 expression in patients with ulcerative colitis may be associated with mucosal inflammatory activity and clinical disease activity through basal cell apoptosis and preventing tissue healing in the pathogenesis of ulcerative colitis.

Topics: Adult; Aged; Antigens, Surface; Apoptosis; Caco-2 Cells; Colitis, Ulcerative; Colon; Female; Flagellin; HT29 Cells; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Milk Proteins; RNA, Messenger; Tumor Necrosis Factor-alpha; Wound Healing; Young Adult

To establish a rat model of ulcerative colitis with syndrome of spleen deficiency and dampness stagnancy.. Sixty rats were divided into normal control group, ulcerative colitis group, ulcerative colitis with syndrome of spleen deficiency and dampness stagnancy group (model group) and strengthening spleen for resolving dampness group. Ulcerative colitis in rats was induced by enema containing trinitrobenzene sulfonic acid (TNBS) and ethanol. The rats in the model group were suffered from standing in water, limiting sleeping time and abnormal diet based on administration of TNBS and ethanol. The rats in the spleen strengthening and dampness resolving group were gastrically administered with Shenlin Baizhu Powder, a compound traditional Chinese herbal medicine. Symptoms, signs and pathological changes in colon tissue of rats were observed after modeling. The levels of interleukin (IL)-6, IL-8 and tumor necrosis factor-α (TNF-α) in serum of rats were measured by enzyme-linked immunosorbent assay.. The rats in the model group showed lethargy, poor appetite, loss of energy, diarrhea and bloody stool. Their body weight decreased significantly compared with the normal control group, and similar changes were found in the comparison of food intake, drinking amount, urine amount, stool wet weight and assay of spontaneous activity (P<0.05). When observed under a light microscope, the colon tissues of rats in the model group showed mucosal edema, congestion, inflammatory cell infiltration and ulceration. The degree of colon injury and IL-6, IL-8 and TNF-α levels were significantly increased (P<0.05) as compared to those in the normal control group. The changes mentioned above were improved by Shenlin Baizhu Powder (P<0.05).. The rat model of ulcerative colitis with syndrome of spleen deficiency and dampness stagnancy is successfully induced and has the characteristics of ulcerative colitis of humans both in pathological changes and in syndrome.

Topics: Animals; Colitis, Ulcerative; Disease Models, Animal; Female; Interleukin-6; Interleukin-8; Male; Medicine, Chinese Traditional; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

To reveal specific gene activation in nitric oxide (NO)-related inflammation we studied differential gene expression in inflammatory bowel disease (IBD).. Total RNA was isolated from 20 biopsies of inflamed mucosa from Crohn's disease (CD) and ulcerative colitis (UC) patients each as well as from six controls, labeled with (32)P-dCTP and hybridized to a human NO gene array. Significant genes were analyzed for functional gene interactions and heatmaps generated by hierarchical clustering. A selection of differentially expressed genes was further evaluated with immunohistochemical staining.. Significant gene expression differences were found for 19 genes in CD and 23 genes in UC compared to controls, both diseases with high expression of ICAM1 and IL-8. Correlation between microarray expression and corresponding protein expression was significant (r = 0.47, p = 0.002). Clustering analysis together with functional gene interaction analysis revealed clusters of coregulation and coexpression in CD and UC: transcripts involved in angiogenesis, inflammatory response mediated by the transcription factor hypoxia-inducible factor 1, and tissue fibrosis. Also, a fourth cluster with transcripts regulated by the transcription factor Sp1 was found in UC.. Expression analysis in CD and UC revealed disease-specific regulation of NO-related genes, which might be involved in perpetuating inflammatory disease activity in IBD.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Case-Control Studies; Chi-Square Distribution; Cluster Analysis; Colitis, Ulcerative; Crohn Disease; Female; Fibrosis; Gene Expression; Gene Expression Profiling; Humans; Hypoxia-Inducible Factor 1; Intercellular Adhesion Molecule-1; Interleukin-1beta; Interleukin-8; Male; Matrix Metalloproteinase 1; Middle Aged; Neovascularization, Pathologic; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Oligonucleotide Array Sequence Analysis; Signal Transduction; Sp1 Transcription Factor; Statistics, Nonparametric; Young Adult

Pentraxin 3 (PTX3) is the first identified long pentraxin, and it is rapidly produced and released by several cell types in response to proinflammatory signals. The aim of this study was to investigate the behavior of neutrophils to produce PTX3 protein in response to proinflammatory cytokine IL-8 in vitro, as well as identify the expression pattern of PTX3 in human ulcerative colitis lesions. Pentraxin 3 protein was found to be present following release upon IL-8 stimulation in cultured neutrophils together with lactoferrin(+)-specific granules localized in neutrophil extracellular traps (NETs) formed by extruded DNA. Neutrophils in colonic mucosal tissue of patients with ulcerative colitis were the main cellular source of PTX3 protein, the expression of which is correlated well with the histological grades of inflammation. Immunofluorescence analysis against anti-lactoferrin antibody revealed the formation of NETs released from neutrophils within crypt abscess lesions, which were found to be activated through the expression of IL-8 receptor B (CXCR2). Of interest, neutrophils depleted of PTX3 protein were displayed, supporting the release of PTX3 from neutrophils in crypt abscess. We suspected that PTX3 protein may contribute to cell-mediated immune defense in inflamed colon tissue, and in particular in crypt abscess lesions, of patients with ulcerative colitis.

Topics: Aged; Antibodies, Monoclonal; Biopsy; C-Reactive Protein; Cells, Cultured; Colitis, Ulcerative; Cytokines; Female; Humans; Inflammation; Interleukin-8; Male; Microscopy, Electron, Scanning; Microscopy, Fluorescence; Middle Aged; Neutrophils; Serum Amyloid P-Component

Recent studies demonstrated that depression was associated with mucosal inflammation in patients with ulcerative colitis (UC). This association had not been studied in patients with UC with ileal pouch-anal anastomosis (IPAA) after restorative proctocolectomy. We hypothesized that depression and mucosal proinflammatory cytokines in UC-patients with pouchitis were associated.. We assessed 18 IPAA-UC-patients with pouchitis and 19 IPAA-UC-patients without pouchitis. Mucosal biopsies were taken from the areas with maximal inflammation in the pouch or from the posterior wall of the pouch if the pouch had a normal endoscopic appearance. Disease activity was assessed by the Pouch Disease Activity Index. The expression of mucosal proinflammatory gene transcripts (interleukin-8 [IL-8] and interleukin-1ß [IL-1b]) was quantified by real-time polymerase chain reaction. Depression was assessed by the Hospital Anxiety and Depression Scale (HADS). Pearson correlations between depression and cytokine transcripts were calculated.. The correlation of HADS depression scores of patients with pouchitis with IL-8 was r=0.51 (p=0.03) and with IL-1ß was r=0.47 (p=0.04). The correlation between the HADS depression scores of patients without pouchitis with IL-8 was r=-0.19; (p=0.21) and with IL-1ß was r=-0.12 (p=0.30).. Depression is associated with mucosal proinflammatory cytokines in patients with pouchitis after restorative proctocolectomy in patients with UC.

Topics: Adult; Colitis, Ulcerative; Cross-Sectional Studies; Depression; Female; Humans; Interleukin-1beta; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Pilot Projects; Polymerase Chain Reaction; Pouchitis; Proctocolectomy, Restorative; Surveys and Questionnaires

Topics: Colitis, Ulcerative; Depression; Female; Humans; Interleukin-1beta; Interleukin-8; Intestinal Mucosa; Male; Pouchitis

Dendritic cell (DC) function is believed to be of critical importance for the pathogenesis of inflammatory bowel disease (IBD). To date, most research in animal models and the few human data available is restricted to myeloid DC, while plasmacytoid DC (pDC) capable of controlling both innate and adaptive immune responses have not yet been investigated systematically in human Crohn's disease (CD) or ulcerative colitis (UC). CD11c(-) , CD303(+) /CD304(+) and CD123(+) pDC from peripheral blood (n = 90), mucosal tissue (n = 28) or mesenteric lymph nodes (n = 40) (MLNs) of patients with UC and CD or controls were purified and cultured. Thereafter, pDC were enumerated, phenotyped and cytokine secretion measured by flow cytometry (FACS), immunohistochemistry and/or cytometric bead array, respectively. Interferon (IFN)-α secretion following cytosine phosphatidyl guanine (CpG) A oligodeoxynucleotide (ODN) 2216 (5'-GGGGGACGATCGTCGGGGGG-3') stimulation was assessed by enzyme-linked immunosorbent assay (ELISA). We found a significantly higher frequency of pDC in the inflamed colonic mucosa and MLN of IBD patients. Moreover, the fraction of CD40 and CD86 expressing cultured peripheral blood pDC was significantly higher in flaring UC and CD patients and their secretion of tumour necrosis factor (TNF)-α, interleukin (IL)-6 and IL-8 were increased significantly compared with controls. In contrast, the IFN-α secretion of peripheral blood pDC isolated from flaring IBD, particularly in UC patients, was reduced significantly compared with controls. Our data suggest an aberrant distribution and function of pDC in IBD, contrary to their generally implicated role as inducers of tolerance. We speculate that the impaired IFN-α secretion may relate to the hypothesized defect in innate immunity in IBD and could also impact upon the generation of regulatory T cells (T(reg) ).

Topics: Adult; Aged; Antigens, CD; Colitis, Ulcerative; Crohn Disease; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Humans; Immunohistochemistry; Interferon-alpha; Interleukin-6; Interleukin-8; Intestinal Mucosa; Lymph Nodes; Male; Middle Aged; Oligodeoxyribonucleotides; Tumor Necrosis Factor-alpha

Chitinase 3-like-1 (CHI3L1/YKL-40) is a protein secreted from restricted cell types including colonic epithelial cells (CECs) and macrophages. CHI3L1 is an inflammation-associated molecule, and its expression is enhanced in persons with colitis and colon cancer. The biological function of CHI3L1 on CECs is unclear. In this study, we investigated the role of CHI3L1 on CECs during the development of colitis-associated neoplasia. We analyzed colonic samples obtained from healthy persons and from persons with ulcerative colitis with or without premalignant or malignant changes. DNA microarray and RT-PCR analyses significantly increased CHI3L1 expression in non-dysplastic mucosa from patients with inflammatory bowel disease (IBD) who had dysplasia/adenocarcinoma compared with that in healthy persons and in patients with IBD who did not have dysplasia. As determined by IHC, CHI3L1 was expressed in specific cell types in the crypts of colonic biopsies obtained from patients with ulcerative colitis who have remote dysplasia. Purified CHI3L1 efficiently activated the NF-κB signaling pathway and enhanced the secretion of IL-8 and TNF-α in SW480 human colon cancer cells. In addition, colon cancer cell proliferation and migration were significantly promoted in response to CHI3L1 in these cells. In summary, CHI3L1 may contribute to the proliferation, migration, and neoplastic progression of CECs under inflammatory conditions and could be a useful biomarker for neoplastic changes in patients with IBD.

Topics: Adipokines; Biomarkers, Tumor; Cell Movement; Cells, Cultured; Chitinase-3-Like Protein 1; Colitis, Ulcerative; Colon; Colorectal Neoplasms; Dose-Response Relationship, Drug; Epithelial Cells; Female; Humans; Interleukin-8; Intestinal Mucosa; Irritable Bowel Syndrome; Lectins; Male; Middle Aged; NF-kappa B; Precancerous Conditions; Tumor Necrosis Factor-alpha

High-mobility group box 1 (HMGB1) is a nuclear protein with functions in the regulation of transcription. In inflammatory conditions, HMGB1 is actively secreted from immune cells in the extracellular matrix, where it behaves as a proinflammatory cytokine. The aim of the present study was to investigate the role of HMGB1 in pediatric inflammatory bowel disease (IBD).. We analyzed the stools of 19 children with Crohn's disease (CD), 21 with ulcerative colitis (UC), and 13 controls. The gene/protein expression levels of HMGB1 were assessed in bioptic specimens of all children using real-time PCR and western blot assay. Finally, intracellular localization of the protein was analyzed by western blot, after separation of nuclear and cytoplasmic extracts, and by immunohistochemistry.. HMGB1 protein levels were significantly increased (P<0.001) in the stools of patients, but were undetectable in the controls; fecal HMGB1 correlated well with fecal calprotectin levels (r: 0.77 in CD, r: 0.70 in UC; P<0.01); and mRNA and protein expression were unchanged in inflamed bioptic tissues compared with controls. However, by separately analyzing the nuclear and cytoplasmic fraction, we detected the cytoplasmic HMGB1 expression to be significantly enhanced (P<0.01) in the inflamed tissues of the patients. In addition, HMGB1 was significantly detected in 16 patients with inactive disease, whose endoscopic scores showed persisting inflammation, suggesting that it may be a sensitive marker of mucosal inflammation, although the disease is clinically inactive.. It was shown for the first time in our study that HMGB1 is secreted by human inflamed intestinal tissues and abundantly found in the stools of IBD patients. Hence, it can be considered as a novel marker for intestinal inflammation. We can also suggest that the presence of HMGB1 in large amounts in the fecal stream of IBD patients is mainly due to active secretion of the protein stored in the nucleus rather than a "de novo" synthesis.

Topics: Active Transport, Cell Nucleus; Adolescent; Biomarkers; Biopsy; Caco-2 Cells; Cell Nucleus; Child; Child, Preschool; Colitis, Ulcerative; Crohn Disease; Feces; Female; HMGB1 Protein; Humans; Infant; Interferon-alpha; Interferon-gamma; Interleukin-8; Intestinal Mucosa; Male; RNA, Messenger

Escherichia coli gene fimA was the most frequent gene that occurred in the intestine of all investigated groups. All subjects with fimA gene had significantly higher values of tumor necrosis factor alpha (TNF-α) and CRP than those with other E. coli genes. There was also a tendency to increased serum interleukin (IL)-6 levels in patients carrying the fimA gene; however, no relation was observed to serum IL-8 and IL-10. Patients with Crohn's disease had significantly higher IL-6 than those with ulcerative colitis (UC) and controls. The highest levels of TNF-α were detected in the UC group. There were no significant differences in serum IL-8 and IL-10 between all three groups. The presence of E. coli gene fimA in the large bowel of patients with IBD is related to the immunological activity of the disease which may be important from the aspect of therapeutical strategy.

Topics: Adult; Aged; Anti-Bacterial Agents; Bacterial Typing Techniques; Case-Control Studies; Colitis, Ulcerative; Crohn Disease; Czech Republic; Escherichia coli; Escherichia coli Infections; Female; Fimbriae Proteins; Humans; Inflammation; Interleukin-10; Interleukin-6; Interleukin-8; Intestines; Male; Middle Aged; Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

Saccharomyces boulardii (Sb) is a probiotic yeast that has demonstrated efficacy in pilot studies in patients with inflammatory bowel disease (IBD). Microbial antigen handling by dendritic cells (DC) is believed to be of critical importance for immunity and tolerance in IBD. The aim was to characterize the effects of Sb on DC from IBD patients. Highly purified (>95%), lipopolysaccharide-stimulated CD1c(+)CD11c(+)CD123(-) myeloid DC (mDC) from patients with ulcerative colitis (UC; n = 36), Crohn's disease (CD; n = 26), or infectious controls (IC; n = 4) were cultured in the presence or absence of fungal supernatant from Sb (SbS). Phenotype and cytokine production and/or secretion of IBD mDC were measured by flow cytometry and cytometric bead arrays, respectively. T cell phenotype and proliferation were assessed in a mixed lymphocyte reaction (MLR) with allogenic CD4(+)CD45RA(+) naïve T cells from healthy donors. Mucosal healing was investigated in epithelial wounding and migration assays with IEC-6 cells. SbS significantly decreased the frequency of CD40-, CD80-, and CD197 (CCR7; chemokine receptor-7)-expressing IBD mDC and reduced their secretion of tumor necrosis factor (TNF)-α and interleukin (IL)-6 while increasing IL-8. In the MLR, SbS significantly inhibited T cell proliferation induced by IBD mDC. Moreover, SbS inhibited T(H)1 (TNF-α and interferon-γ) polarization induced by UC mDC and promoted IL-8 and transforming growth factor-β-dependent mucosal healing. In summary, we provide novel evidence of synergistic mechanisms how Sb controls inflammation (inhibition of T cell costimulation and inflammation-associated migration and mobilization of DC) and promotes epithelial restitution relevant in IBD.

Topics: B7-1 Antigen; CD40 Antigens; Cell Division; Cell Movement; Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Dendritic Cells; Female; Humans; Immunotherapy; Interleukin-6; Interleukin-8; Lymphocyte Culture Test, Mixed; Male; Probiotics; Receptors, CCR7; Saccharomyces; T-Lymphocytes; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Reduced alpha-defensin expression has been reported in the terminal ileum (TI) of adult patients with ileal Crohn's disease (CD). However, little is known about alpha-defensin expression in children with chronic inflammatory bowel disease (IBD).. In all, 283 intestinal biopsies were obtained from children with CD, ulcerative colitis (UC), and healthy controls. Absolute mRNA copy numbers for HD5, HD6, IL-8, Villin 1, and Tcf-4 were analyzed by reverse-transcription polymerase chain reaction (RT-PCR). HD5 immunostaining was performed on biopsy sections and patients genotyped for NOD2 mutations.. Equal expression levels of alpha-defensins (HD5 and HD6) were found in TI biopsies of children with ileal CD (L1+L3) compared to patients with colonic disease (L2) and healthy controls. In contrast, we found significantly higher levels of alpha-defensins in the TI of children with UC compared to CD and controls. Reduced expression of Tcf-4 was observed exclusively in the duodenum and TI of CD patients with L1+L3 phenotype. We demonstrate significantly increased expression of HD5 and HD6 in the inflamed colon of IBD children (UC and CD) attributable to the presence of metaplastic Paneth cells.. In this study no difference in alpha-defensin expression was found in the TI of CD children and controls. However, significant reduction of Tcf-4 in L1+L3 phenotype suggests that a possibly impaired PC differentiation may lead to altered HD5 and HD6 expression at some stage of disease. Additionally, substantially increased expression of alpha-defensins in the inflamed colonic mucosa of children with IBD raises the question for their potential involvement in modulating inflammation in these patients.

Topics: alpha-Defensins; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; Case-Control Studies; Child; Colitis, Ulcerative; Colon; Crohn Disease; Female; Fluorescent Antibody Technique; Humans; Ileum; Immunoenzyme Techniques; Interleukin-8; Intestinal Mucosa; Male; Microfilament Proteins; Paneth Cells; Prognosis; Prospective Studies; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription Factor 4; Transcription Factors

Endoplasmic reticulum (ER) stress has been suggested to play a role in inflammatory bowel disease (IBD). The three branches (ATF6, IRE1 and PERK) of the unfolded protein response (UPR) have different roles and are not necessarily activated simultaneously.. Expression of UPR-related genes was investigated in colonic and ileal biopsies of 23 controls, 15 ulcerative colitis (UC) and 54 Crohn's disease (CD) patients. This expression was confirmed at protein level in colonic and ileal samples of five controls, UC and CD patients. HSPA5, PDIA4 and XBP1s were significantly increased in colonic IBD at mRNA and/or protein levels, indicating activation of the ATF6 and IRE1 branch. Colonic IBD was associated with increased phosphorylation of EIF2A suggesting the activation of the PERK branch, but subsequent induction of GADD34 was not observed. In ileal CD, no differential expression of the UPR-related genes was observed, but our data suggested a higher basal activation of the UPR in the ileal mucosa of controls. This was confirmed by the increased expression of 16 UPR-related genes as 12 of them were significantly more expressed in ileal controls compared to colonic controls. Tunicamycin stimulation of colonic and ileal samples of healthy individuals revealed that although the ileal mucosa is exhibiting this higher basal UPR activation, it is still responsive to ER stress, even more than colonic mucosa.. Activation of the three UPR-related arms is seen in colonic IBD-associated inflammation. However, despite EIF2A activation, inflamed colonic tissue did not increase GADD34 expression, which is usually involved in re-establishment of ER homeostasis. This study also implies the presence of a constitutive UPR activation in healthy ileal mucosa, with no further activation during inflammation. Therefore, engagement of the UPR differs between colon and ileum and this could be a factor in the development of ileal or colonic disease.

Topics: Activating Transcription Factor 6; Adolescent; Adult; Aged; Case-Control Studies; Child; Colitis, Ulcerative; Colon; Crohn Disease; Endoplasmic Reticulum Chaperone BiP; Endoplasmic Reticulum Stress; Endoribonucleases; Endoscopy, Gastrointestinal; Heat-Shock Proteins; Humans; Ileum; Interleukin-8; Intestinal Mucosa; Male; Membrane Proteins; Middle Aged; Protein Serine-Threonine Kinases; RNA, Messenger; Transcriptome; Tunicamycin; Unfolded Protein Response; Young Adult

Cytokines have validated roles in the immunopathogenesis of inflammatory bowel disease (IBD). This study was to investigate the expressions of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-8, and IL-10 mRNAs in the colonic mucosa of patients with ulcerative colitis (UC) during active and quiescent UC.. At colonoscopy, biopsies were taken from inflamed and non-inflamed mucosa of patients with steroid-naive UC (n = 15), non-IBD inflammatory colitis controls (ICC, n = 6), and non-colitis controls (NCC, n = 14). The presence of extensive mononuclear cells and neutrophils infiltrate in the lamina propria, cryptitis, and epithelial damage defined an inflammatory lesion in the mucosa. Quantitative cytokine mRNA expressions in biopsies were measured by real-time polymerase chain reaction (PCR).. Of 15 UC patients, 3 remitted with 5-aminosalicylate and 11 received granulocytapheresis; of these, 10 remitted. At baseline, IL-6, IL-8, TNF-alpha, and IL-10 mRNAs were high in inflamed mucosa compared with NCC (P < 0.01). In active UC, IL-6, IL-8 and IL-10 mRNAs were high compared with non-inflamed mucosa (P = 0.03, P = 0.03, P < 0.05, respectively). Both TNF-alpha mRNA (P = 0.03) and IL-6 mRNA (P = 0.04) were higher in UC compared with ICC. Even in non-inflamed mucosa, IL-8 and TNF-alpha mRNA expressions were high compared with NCC. Both IL-6 and IL-8 mRNAs decreased to normal levels after granulocytapheresis.. During active UC, all 4 cytokine mRNA levels were high; only IL-6 and IL-8 mRNAs decreased to normal levels during remission. IL-8 mRNA was high even at sites of endoscopically quiescent UC during active disease. Steroid naïve patients respond well to granulocytapheresis.

Topics: Administration, Oral; Adult; Anti-Inflammatory Agents, Non-Steroidal; Biopsy; Colitis, Ulcerative; Colon; Colonoscopy; Cytokines; Dose-Response Relationship, Drug; Drug Resistance; Female; Follow-Up Studies; Gene Expression Regulation; Granulocytes; Humans; Interleukin-6; Interleukin-8; Intestinal Mucosa; Leukapheresis; Male; Mesalamine; Middle Aged; Polymerase Chain Reaction; Prognosis; Remission Induction; Retrospective Studies; RNA, Messenger; Tumor Necrosis Factor-alpha; Young Adult

Growing evidence indicates that innate immunity, including toll-like receptor (TLR) signalling, plays a role in inflammatory bowel disease (IBD). This may also apply in the case of TLR-8, which has recently been shown to reverse the immunosuppressive function of regulatory T cells. However, the role of TLR-8 in IBD is currently unknown, and therefore we investigated the expression of TLR-8 and its natural antagonist, Tollip, in normal and inflamed human gut, and examined whether the receptor is functionally active.. TLR-8 and Tollip mRNA expression were measured in colonic epithelial cells (CEC) and lamina propria mononuclear cells (LPMNC) by quantitative polymerase chain reaction. TLR-8 protein expression was visualized in whole biopsy specimens by indirect immunofluorescence microscopy. Cellular localization of TLR-8 protein was assessed by immuno-electron microscopy. IL-8 secretion was measured by ELISA after stimulation with TLR-8 ligand.. TLR-8 mRNA and protein expression were substantially up-regulated in CEC from inflamed mucosa from patients with ulcerative colitis (approximately 350-fold, p<0.01) and Crohn's disease (approximately 45-fold, p<0.05) compared to controls. TLR-8 proteins resided on the luminal surface membrane and in intracellular organelles. Tollip was not increased in CEC from IBD patients. CEC from normal mucosa responded to TLR-8 stimulation by secreting IL-8. TLR-8 was expressed only on the mRNA level in LPMNC with no differences between IBD patients and controls.. Expression of TLR-8, but not Tollip, is highly up-regulated in the colonic epithelium from patients with active IBD. Since the receptor is functionally active, our data suggest that TLR-8 signalling is important in the pathogenesis of IBD.

Topics: Adolescent; Adult; Aged; Colitis, Ulcerative; Crohn Disease; Epithelial Cells; Female; Gene Expression; Humans; Inflammatory Bowel Diseases; Interleukin-8; Intestinal Mucosa; Intracellular Signaling Peptides and Proteins; Male; Middle Aged; Toll-Like Receptor 8; Up-Regulation

Interleukin-8 (IL-8), a CXC chemokine that recruits and activates inflammatory cells, plays a critical role in the pathogenesis of ulcerative colitis (UC). There are no studies on the association of single nucleotide polymorphisms (SNPs) of the IL-8 gene with the risk of UC.. All 162 unrelated UC patients and 203 control subjects were analyzed for 5 IL-8 SNPs ((-845 (T/C), -738 (T/A), -353 (A/T), -251 (T/A) and +678 (T/C)) using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay and PCR-sequence-specific primers (SSP) method. Serum IL-8 concentrations were measured in all subjects.. Individual SNPs were not associated with risk for UC. However, the frequency of -353A/-251A/+678T haplotype was significantly higher in UC patients than in healthy controls (OR=1.454, p=0.036). By subgroup analyses, this haplotype tended to be more common in severe UC patients than in those with mild-to-moderate disease (OR=2.281, p=0.027). Furthermore, patients with AAT diplotype showed significantly increased serum IL-8 concentrations than those with other diplotypes (p<0.001).. These results suggest that IL-8 is a novel susceptibility gene to UC in Chinese UC patients, and furthermore, that IL-8 polymorphisms may be related to severe clinical subtype of UC.

Topics: Adult; Aged; Alleles; Asian People; Colitis, Ulcerative; Female; Genotype; Health; Humans; Interleukin-8; Male; Middle Aged; Polymorphism, Genetic; Risk Factors

Interleukin 2 (IL-2)- and IL-10-knockout mice develop spontaneous colitis under conventional but not germ-free conditions, suggesting that commensal bacteria play an important role in the pathogenesis of colitis. However, interactions between commensal bacteria and colonic epithelial cells have not been fully investigated. We therefore assessed the ability of various commensal bacteria and probiotics to adhere to and invade colonic epithelial cells. Effects of the bacteria on production of proinflammatory cytokines were also measured. Commensal bacteria, including mucosal organisms isolated from ulcerative colitis (UC) patients, such as Fusobacterium varium, reported as a possible pathogen in UC, Bacteroides vulgatus, Escherichia coli and Clostridium clostridioforme, as well as their type strains and probiotics, were assessed for their ability to adhere to and invade colonic epithelial cells using two cell lines, SW-480 and HT-29. Our experiments employed co-incubation, a combination of scanning and transmission electron microscopy and recovery of bacteria from infected-cell lysates. F. varium and several other commensal bacteria, but not probiotics, adhered to colonic epithelial cells and invaded their cytoplasm. ELISA and real-time PCR revealed that the host cells, particularly those invaded by F. varium, showed significant increases in IL-8 and TNF-alpha concentrations in supernatants, with elevation of IL-8, TNF-alpha, MCP-1 and IL-6 mRNAs. Furthermore, IL-8 and TNF-alpha expression and nuclear phosphorylated NF-kappaB p65 expression could be immunohistochemically confirmed in inflamed epithelium with cryptitis or crypt abscess in UC patients. Certain commensal bacteria can invade colonic epithelial cells, activating early intracellular signalling systems to trigger host inflammatory reactions.

Topics: Adenocarcinoma; Animals; Bacterial Adhesion; Cell Line, Tumor; Colitis, Ulcerative; Colon; Colonic Neoplasms; Cytokines; DNA Primers; Humans; Inflammation; Interleukin-10; Interleukin-2; Interleukin-8; Intestinal Mucosa; Mice; Mice, Knockout

Topics: Adsorption; Antibodies; Blood Component Removal; Colitis, Ulcerative; Granulocytes; Head; Humans; Interleukin-6; Interleukin-8; Male; Monocytes; Phosphatidylserines; Prothrombin; Pyoderma Gangrenosum; Young Adult

The effects of Wumeiwan (WMW) on TNF-alpha, IL-6, IL-8, IL-10 and NF-kappaBp65 in rats with ulcerative colitis (UC) were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine (SASP) was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley (SD) rats were randomly divided into four groups (n=14 in each group, with equal ratio of male and female): normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene (DNCB) immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-alpha, IL-10 and the expression of NF-kappaBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-alpha were significantly increased (P<0.01), while those of IL-10 significantly reduced (P<0.01) after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-alpha were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group (P<0.05). The levels of IL-6, IL-8, and TNF-alpha were lower, while the level of IL-10 was higher in WMW group than in SASP group. NF-kappaBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-kappaBp65 in WMW group and SASP group was obviously lower than in model group (P<0.01), and there was significant difference between WMW group and SASP group (P<0.05). It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-alpha, IL-6, IL-8, and inhibit the NF-kappaBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.

Topics: Animals; Colitis, Ulcerative; Colon; Cytokines; Drugs, Chinese Herbal; Female; Interleukin-10; Interleukin-6; Interleukin-8; Male; Phytotherapy; Random Allocation; Rats; Rats, Sprague-Dawley; Transcription Factor RelA; Tumor Necrosis Factor-alpha

The modulation of the intestinal expression of detoxifying proteins by relevant transcription factors, intracellular receptors and cytokines in ulcerative colitis (UC) is poorly understood. Here, we compared the intestinal expression of drug transporters, metabolizing enzymes and putative regulatory genes between inflamed and noninflamed tissue and studied their modulation by disease activity.. Sigmoidal biopsies of 18 UC patients and 18 healthy volunteers matched for age, gender and ABCB1 3435C>T genotype were investigated for mRNA expression levels of 43 systematically selected candidate genes by low-density array real-time PCR. Additionally, the ABCB1 gene product P-glycoprotein was visualized by immunohistochemistry and quantified by western blotting. Disease phenotype was categorized by clinical, endoscopic and histopathological examination. Disease activity was quantified by clinical activity index.. In inflamed sigmoidal tissue from UC patients, 11 genes (NAT1, NR2B1, CEBPB, IFG, IL8, IL10, S100A12, SPP1, DEFA5, DEFA6 and HAMP) were overexpressed. By contrast, only the major human efflux transporter ABCB1 showed significantly lower expression levels, that were inversely correlated with those of certain antimicrobial peptides (DEFA5/6) and cytokines (IL1beta and IL8). Cell culture experiments revealed a time-dependent decrease of ABCB1 expression upon IL8 exposure. Disease activity profoundly modified ABCB1 expression, indicated by an inverse correlation of clinical activity index values with ABCB1 mRNA expression (r = -0.603; p = 0.017) and markedly reduced protein expression in UC patients with moderate and severe symptomology (p = 0.011).. Cytokine-mediated downregulation of the major human efflux transporter ABCB1 in inflamed intestine of UC patients is presumably dependent on disease activity, with a possible contribution from IL8.

Topics: Adult; ATP Binding Cassette Transporter, Subfamily B; ATP Binding Cassette Transporter, Subfamily B, Member 1; Blotting, Western; Case-Control Studies; Cell Line, Tumor; Colitis, Ulcerative; Colon, Sigmoid; Down-Regulation; Gene Expression Profiling; Genotype; Humans; Immunity, Innate; Immunohistochemistry; Interleukin-8; Polymorphism, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Severity of Illness Index

Angiogenesis is required for wound healing and repair, but dysregulated angiogenesis is involved in gastrointestinal inflammation. Bacillus polyfermenticus (B.P.) is a probiotic bacterium clinically used for a variety of intestinal disorders in East Asia. Here we investigated the effect of B.P. on angiogenesis of human intestinal microvascular endothelial cells (HIMECs) and wound healing in intestinal mucosa. Exposure of HIMECs to the conditioned medium of B.P. cultures (B.P. CM) increased cell migration, permeability, and tube formation. Production of the proangiogenic cytokine IL-8 was increased by B.P. CM, and neutralizing antibodies against IL-8 or IL-8 receptor CXCR2 reduced tube formation as well as actin stress fiber formation. B.P. CM also increased NF-kappaB activation, and inhibitors of NF-kappaB suppressed B.P. CM-induced tube formation and IL-8 production. Furthermore, B.P. facilitated recovery of mice from colitis as shown by increased body weight and reduced rectal bleeding and histological severity. B.P. also increased angiogenesis and mouse IL-8 production in the mucosal layer. Collectively, these results show that B.P. increases angiogenesis of HIMECs in a NF-kappaB/IL-8/CXCR2-dependent manner. Moreover, B.P. promotes angiogenesis in the mucosa during recovery of mice from colitis, suggesting that this probiotic may be clinically used to facilitate intestinal wound healing.

Topics: Animals; Bacillus; Capillary Permeability; Cell Movement; Cell Proliferation; Cells, Cultured; Colitis, Ulcerative; Culture Media, Conditioned; Endothelial Cells; Gene Expression; Humans; Interleukin-8; Intestines; Male; Mice; Mice, Inbred Strains; Neovascularization, Physiologic; NF-kappa B; Phenylurea Compounds; Phosphorylation; Probiotics; Receptors, Interleukin-8B; Stress Fibers; Vascular Endothelial Growth Factor A; Wound Healing

Pouchitis after total rectocolectomy is among the most common complications of patients with ulcerative colitis (UC). However, its frequency is quite rare in patients with familial adenomatous polyposis (FAP). We evaluated the inflammatory and pro-apoptotic activity in endoscopically normal mucosa of the ileal pouch in patients with UC and FAP.. Twenty patients (10 with UC and 10 with FAP) with "J" pouch after total proctocolectomy were studied as were 10 normal controls. Biopsies were obtained from the mucosa of the pouch of UC and FAP patients and from the normal ileum of controls. The expression levels of TNF-alpha, IL-1beta, IL-6, IL-8 and phospho-BAD were determined by immunoblotting. Activated NFkappaB was evaluated by immuno-precipitation and immunoblotting for IkappaB kinase beta.. Patients with UC had higher levels of IL-1beta, IL-6, IL-8 and TNF-alpha than patients with FAP. The level of TNF-alpha was higher in patients with UC than in patients with FAP; both patient groups had TNF-alpha levels higher than controls. Activation of NFkappaB was similar in all three groups. The expression of phospho-BAD was significantly lower in patients with FAP than in patients with UC.. As compared with patients with FAP, patients with UC presented increased levels of some pro-inflammatory cytokines, even in the absence of clinical or endoscopic signs of pouchitis. Patients with FAP presented lower levels of pro-inflammatory proteins and of phospho-BAD. These findings may explain the higher rates of progression to pouchitis in UC patients, which could correlate with mucosal atrophy that occurs in inflamed tissue.

Topics: Adenomatous Polyposis Coli; Adult; Analysis of Variance; Apoptosis Regulatory Proteins; bcl-Associated Death Protein; Colitis, Ulcerative; Colonic Pouches; Cytokines; Female; Humans; Immunoblotting; Interleukin-1; Interleukin-6; Interleukin-8; Male; Middle Aged; NF-kappa B; Pouchitis; Proctocolectomy, Restorative; Tumor Necrosis Factor-alpha

Ulcerative colitis (UC) mainly affects the colon and rectum, whereas the chronic inflammatory process in Crohn's disease (CD) can affect any part of the gastrointestinal tract. Recently, however, upper gastrointestinal lesions have been reported in UC patients. In this study, we investigated the immunological status of the stomach in UC or CD patients.. 26 patients each with UC, CD or functional dyspepsia (FD) underwent diagnostic upper gastrointestinal endoscopy, and biopsy specimens were obtained from the gastric antrum. The contents of interleukin (IL)-6 and IL-8 in the organ culture supernatants of antral mucosal tissues were measured with enzyme-linked immunosorbent assay.. Endoscopically abnormal findings in the stomach were more frequent in CD than in UC or FD patients. Mononuclear cell infiltration and IL-6 production in the gastric antrum did not significantly differ between UC and FD patients, but were higher in those with CD. There was no significant difference in neutrophil infiltration or IL-8 production between UC, CD, and FD patients.. UC patients did not show the immunological abnormalities in the stomach seen in CD patients.

Topics: Adolescent; Adult; Colitis, Ulcerative; Crohn Disease; Endoscopy, Gastrointestinal; Female; Gastric Mucosa; Humans; Interleukin-6; Interleukin-8; Male; Neutrophil Infiltration; Organ Culture Techniques; Stomach

To measure the serum levels of neutrophils chemokine granulocyte chemotactic protein-2 (GCP-2) and interleukin-8 (IL-8) in Crohn's disease (CD) and ulcerative colitis (UC) patients and compare them with serum levels of growth-related oncogene (GRO-alpha).. Forty-two patients with inflammatory bowel disease (24 CD and 18 UC) and 38 matched healthy subjects were recruited. Their serum GCP-2, IL-8 and GRO-alpha were measured by a specific enzyme immunoassay kit.. The serum levels of GCP-2 were significantly higher in the CD than the UC patients but lower than in the healthy subjects. The GCP-2 in the UC patients were significantly lower than in the healthy subjects. The GRO-alpha levels were significantly higher in the IBD patients than in the healthy subjects. The IL-8 levels were under the detectable limit in both the IBD and the healthy subjects.. In this group of patients, GCP-2 did not participate in the inflammatory response in IBD. GRO-alpha could be an important factor that enhances the inflammatory state in IBD.

Topics: Adult; Chemokine CXCL1; Chemokine CXCL6; Colitis, Ulcerative; Crohn Disease; Humans; Immunoenzyme Techniques; Interleukin-8

Activation of nuclear factor-kappa B (NF-kappaB), which controls transcription of various proinflammatory cytokine genes, has been shown to play a critical role in the pathogenesis of ulcerative colitis (UC). The aim of this study was to investigate if NF-kappaB p65 antisense oligonucleotides may affect the expression of NF-kappaB p65 and cytokines in lamina propria mononuclear cells (LPMCs) from patients with UC.. LPMCs, which were isolated from intestinal mucosal biopsy specimens from patients with UC, were cultured with or without NF-kappaB p65 antisense oligonucleotides, missense oligonucleotides and dexamethasone. NF-kappaB p65 expression was determined by Western blot analysis. The expression of cytokine mRNA was studied by reverse transcription-polymerase chain reaction (RT-PCR). Cytokine levels were measured by enzyme-linked immunosorbent assay.. NF-kappaB p65 antisense oligonucleotides resulted in downregulation of NF-kappaB p65 expression, blocked the expression of IL-1beta mRNA and IL-8 mRNA, and strikingly reduced the production of IL-1beta and IL-8. These effects were greater than those of dexamethasone in cultured LPMCs from patients with UC (p <0.05).. Application of NF-kappaB p65 antisense oligonucleotides may serve as a novel molecular approach for the treatment of patients with UC.

Topics: Adult; Anti-Inflammatory Agents; Colitis, Ulcerative; Dexamethasone; Female; Humans; Interleukin-1beta; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Mucous Membrane; Oligonucleotides, Antisense; Transcription Factor RelA

Currently, published reports of mucosal inflammation in the terminal ileum of ulcerative colitis (UC) before colectomy are scarce.. To investigate inflammation in the terminal ileum of UC patients by endoscopic examinations and measurement of mucosal cytokine profiles.. Fifty consecutive patients with active UC were studied. At ileocolonoscopy, mucosal biopsies were taken from the terminal ileum. As control, mucosal biopsies from 20 patients without inflammation were examined.. Thirty-eight patients showed endoscopically normal terminal ileum, four showed backwash ileitis, and eight showed non-backwash ileitis (ileitis with normal caecum). Pancolitis was observed in all of four patients with backwash ileitis, in 4 of 8 (50%) with non-backwash ileitis, and in 4 of 38 (11%) without ileal inflammation (P=0.0002). Extraintestinal manifestations were observed in none of 4 patients with backwash ileitis, in 6 of 8 (75%) with non-backwash ileitis, and in 3 of 38 (8%) without ileal inflammation (P<0.0001). In patients with backwash ileitis and non-backwash ileitis, ileal interleukin [IL]-1beta, IL-6, IL-8 and tumour necrosis factor-alpha levels were significantly elevated compared with the control group. Only extraintestinal manifestation was associated with higher ileal cytokine levels, whereas age, sex, and duration, extent and severity of UC did not show any apparent association.. In patients with backwash ileitis, elevated ileal cytokines might reflect a reaction to regurgitation of colonic content into the ileum, but in patients without backwash ileitis, alternative factors are expected to contribute to the aetiology of ileal inflammation. Patients with extraintestinal manifestations had elevated ileal cytokine levels.

Topics: Adult; Colitis, Ulcerative; Cytokines; Endoscopy, Gastrointestinal; Female; Humans; Ileum; Interleukin-1beta; Interleukin-6; Interleukin-8; Intestinal Mucosa; Male; Prospective Studies; Tumor Necrosis Factor-alpha

To determine the anti-inflammatory activity of probiotic Bifidobacteria in Bifidobacteria-fermented milk (BFM) which is effective against active ulcerative colitis (UC) and exacerbations of UC, and to explore the immunoregulatory mechanisms.. Peripheral blood mononuclear cells (PBMNC) from UC patients or HT-29 cells were co-cultured with heat-killed probiotic bacteria or culture supernatant of Bifidobacterium breve strain Yakult (BbrY) or Bifidobacterium bifidum strain Yakult (BbiY) to estimate the amount of IL-10 or IL-8 secreted.. Both strains of probiotic Bifidobacteria contained in the BFM induced IL-10 production in PBMNC from UC patients, though BbrY was more effective than BbiY. Conditioned medium (CM) and DNA of both strains inhibited IL-8 secretion in HT-29 cells stimulated with TNF-alpha, whereas no such effect was observed with heat-killed bacteria. The inhibitory effect of CM derived from BbiY was greater than that of CM derived from BbrY. DNAs of the two strains had a comparable inhibitory activity against the secretion of IL-8. CM of BbiY induced a repression of IL-8 gene expression with a higher expression of IkappaB-zeta mRNA 4 h after culture of HT-29 cells compared to that in the absence of CM.. Probiotic Bifidobacterium strains in BFM enhance IL-10 production in PBMNC and inhibit IL-8 secretion in intestinal epithelial cells, suggesting that BFM has anti-inflammatory effects against ulcerative colitis.

Topics: Bifidobacterium; Cell Culture Techniques; Colitis, Ulcerative; Culture Media, Conditioned; Cytokines; DNA, Bacterial; HT29 Cells; Humans; Interleukin-10; Interleukin-8; Leukocytes, Mononuclear; Probiotics; Reverse Transcriptase Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

Various therapies are used for inflammatory bowel diseases (IBD), though none seem to be extremely effective. AP-1 is a major transcription factor that upregulates genes involved in immune and proinflammatory responses. We investigated decoy oligodeoxynucleotide (ODN) targeting AP-1 to prevent dextran sulfate sodium (DSS)-induced colitis in mice. Functional efficacies of synthetic decoy and scrambled ODNs were evaluated in vitro by a reporter gene luciferase assay and measuring flagellin-induced IL-8 expression by HCT-15 cells transfected with ODNs. Experimental colitis was induced in mice with a 2.5% DSS solution in drinking water for 7 days, and decoy or scrambled ODNs were intraperitoneally injected from days 2 to 5. Colitis was assessed by weight loss, colon length, histopathology, and detection of myeloperoxidase (MPO), IL-1beta, and TNF-alpha in colon tissue. Therapeutic effects of AP-1 and NF-kappaB decoy ODNs were compared. Transfection of AP-1 decoy ODN inhibited AP-1 transcriptional activity in reporter assays and flagellin-induced IL-8 production in vitro. In mice, AP-1 decoy ODN, but not scrambled ODN, significantly inhibited weight loss, colon shortening, and histological inflammation induced by DSS. Further, AP-1 decoy ODN decreased MPO, IL-1beta, and TNF-alpha in colonic tissue of mice with DSS-induced colitis. The AP-1 decoy therapeutic effect was comparable to that of NF-kappaB decoy ODN, which also significantly decreased intestinal inflammation. Double-strand decoy ODN targeting AP-1 effectively attenuated intestinal inflammation associated with experimental colitis in mice, indicating the potential of targeting proinflammatory transcription factors in new therapies for IBD.

Topics: Animals; Cell Culture Techniques; Cell Line, Tumor; Colitis, Ulcerative; Colonic Neoplasms; Dextran Sulfate; Flow Cytometry; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Genes, Reporter; Humans; Inflammation; Interleukin-8; Luciferases; Mice; Mice, Inbred BALB C; Oligonucleotides; RNA, Messenger; Specific Pathogen-Free Organisms; Transcription Factor AP-1; Transcription, Genetic; Transfection

Dysregulation of interleukin-8 (IL-8) production has been proposed to contribute to intestinal inflammation in inflammatory bowel disease (IBD) patients. Previous studies, which evaluate adult patients with long-standing or steroid-modulated disease, have reported conflicting results regarding the role of IL-8 in IBD pathogenesis. The present study evaluates IL-8 in colonic organ cultures and sera of newly and previously diagnosed pediatric IBD patients with various degrees of histopathologic activity. Colon and terminal ileum biopsies were obtained from 26 patients with Crohn's disease, 12 with ulcerative colitis, 4 with indeterminate colitis, and 12 age-matched normal controls. IBD patients were additionally characterized as newly or previously diagnosed. Supernatants from organ-cultured lamina propria biopsies and sera were evaluated by ELISA for IL-8 protein. IL-8 increased with degree of histologic inflammation regardless of diagnosis (no pathologic diagnosis, 62.6 ng/ml, interquartile range [IQR] 30.4-94.6 ng/ml; mild, 92.0 ng/ml, IQR 21.9-170.0 ng/ml; moderate, 676.2 ng/ml, IQR 46.4-2967.7 ng/ml; severe, 585.6 ng/ml, IQR 149.7-1602.2 ng/ml; P < 0.01). Lamina propria IL-8 was significantly elevated in moderately and severely inflamed tissue segments (603.26 ng/ml; IQR, 72.15-2240.4 ng/ml) compared to noninflamed and mildly inflamed segments (67.70 ng/ml; IQR, 30.38-124.1 ng/ml; P = 0.0009). There was no significant trend in IL-8 concentration when compared by clinical diagnosis. No significant difference was found in IL-8 concentrations in organ cultures from newly diagnosed patients versus those from previously diagnosed patients. There was no significant correlation between serum IL-8 concentration and organ culture IL-8 concentration. We conclude that higher concentrations of IL-8 are found in more histologically inflamed tissue segments from pediatric IBD patients. IL-8 does not appear to be associated with clinical IBD subtype. IL-8 appears to be an integral part of both early and established mucosal inflammation in pediatric IBD patients. These findings suggest that IL-8-specific therapies may universally modify inflammatory activity in IBD patients.

Topics: Adolescent; Adult; Case-Control Studies; Child; Child, Preschool; Colitis, Ulcerative; Colon; Crohn Disease; Enzyme-Linked Immunosorbent Assay; Female; Humans; Ileum; Inflammatory Bowel Diseases; Interleukin-8; Intestinal Mucosa; Linear Models; Male; Mucous Membrane; Organ Culture Techniques; Philadelphia; Severity of Illness Index

Urocortin II (UcnII) is a neuropeptide that binds with high affinity to the corticotropin-releasing hormone receptor 2 (CRHR2) in peripheral tissues. UcnII is synthesised in the intestine, but its role in human intestinal inflammation is largely unknown.. Responses of human colonic epithelial cells expressing CRHR2 to stimulation by UcnII were measured using ELISA, western blot analysis, real-time reverse transcription-PCR (RT-PCR) and interleukin (IL)8 promoter activity. Expression levels of CRHR2 and UcnII in human colitis were determined by immunofluorescence and real-time RT-PCR in mucosal biopsies from patients with Crohn's and ulcerative colitis, and in human intestinal xenografts after exposure to Clostridium difficile toxin A.. It is reported here that expression of CRHR2 mRNA and protein in human colonic epithelial cells (HT-29) are increased by exposure to C difficile toxin A or tumour necrosis factor (TNF)alpha. Stimulation of non-transformed NCM460 colonocytes overexpressing CRHR2alpha receptor with UcnII resulted in a time- and concentration-dependent increase in IL8 production. UcnII stimulation also led to activation of nuclear factor-kappaB (NF-kappaB) and mitogen-acivated protein (MAP) kinase in these cells, as evidenced by degradation of IkappaBalpha and phosphorylation of the p65 subunit of NF-kappaB and extracellularly regulated kinase (ERK) 1/2. Furthermore, expression of UcnII and CRHR2 mRNA was increased in mucosal samples of patients with inflammatory bowel disease, and after exposure of human intestinal xenografts to C difficile toxin A.. These results suggest that UcnII has pro-inflammatory effects in human intestinal cells via the CRHR2alpha receptor and may play an important role in the pathophysiology of colitis in humans.

Topics: Animals; Bacterial Toxins; Cell Line; Colitis; Colitis, Ulcerative; Colon; Corticotropin-Releasing Hormone; Crohn Disease; Enterotoxins; Epithelial Cells; Gene Expression Regulation; Humans; Interleukin-8; Intestines; Mice; Mice, SCID; Mitogen-Activated Protein Kinases; NF-kappa B; Receptors, Corticotropin-Releasing Hormone; RNA, Messenger; Transplantation, Heterologous; Tumor Necrosis Factor-alpha; Urocortins

Topics: Aged, 80 and over; Anti-Inflammatory Agents, Non-Steroidal; Breast Diseases; Colitis, Ulcerative; Female; Humans; Interleukin-8; Mesalamine; Pyoderma Gangrenosum; Treatment Outcome

Relapse of ulcerative colitis is difficult to predict by routine colonoscopy. A high-resolution video-magnifying colonoscope with chromoscopy enables the observation of colorectal mucosal pit patterns.. To investigate the association of pit patterns as assessed by magnifying colonoscopy (MCS) with histological inflammation and mucosal chemokine activity in patients with quiescent ulcerative colitis, and to prospectively analyse the prognostic factors that may predict exacerbations.. MCS was performed in 113 patients with ulcerative colitis in remission. Pit patterns in the rectal mucosa were classified into four MCS grades on the basis of size, shape and arrangement. Mucosal interleukin (IL) 8 activity was measured in biopsy specimens of rectal mucosa and the specimens were assessed for histological disease activity. The patients were then followed until relapse or for a maximum of 12 months. Multivariate survival analysis was carried out to determine the independent predictors of clinical relapse.. A positive correlation was identified between MCS grade, histological grade (p = 0.001) and mucosal IL8 activity (p<0.001). Multivariate proportional hazard model analysis showed that MCS grade was a significant predictor of relapse (relative risk 2.06, p = 0.001). Kaplan-Meier estimate of relapse during 12 months of follow-up was found to increase with increasing MCS grade, with values of 0% for grade 1, 21% for grade 2, 43% for grade 3 and 60% for grade 4.. MCS grading is associated with the degree of histological inflammation and mucosal IL8 activity in patients with quiescent ulcerative colitis, and may predict the probability of subsequent disease relapse in patients with ulcerative colitis in remission.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Colitis, Ulcerative; Colonoscopy; Female; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Proctitis; Prognosis; Prospective Studies; Rectum; Recurrence

Macrophage migration inhibitory factor (MIF) is a cytokine that has potent anti-steroid effects and might be implicated in the pathogenesis of Ulecrative colitis (UC). We defined the functional role of MIF in the glucocorticoid (GC)-resistant inflammatory response in UC. Twenty-four colonic samples were obtained from GC responsive cases, GC refractory cases, Crohn's disease and controls. LPMC were isolated from surgical specimens. MIF was strongly expressed at mRNA levels in refractory cases rather than responsive cases with UC and controls. IL-8 production from LPMC was significantly reduced by GC addition in responsive cases but not in refractory cases. In refractory cases, anti-MIF Ab ameliorated GC-resistant IL-8 production and p38-MAPK activity of LPMC. In addition, p38-MAPK antagonist SB230580 also ameliorated GC-resistant IL-8 production. These results suggest that MIF has an additional proinflammatory activity through the p38-MAPK pathway in GC-resistant UC.

Topics: Adolescent; Adult; Aged; Biopsy; Child; Colitis, Ulcerative; Drug Resistance; Enzyme-Linked Immunosorbent Assay; Female; Glucocorticoids; Humans; Imidazoles; Immunohistochemistry; Interleukin-8; Intestinal Mucosa; Macrophage Migration-Inhibitory Factors; Male; Middle Aged; p38 Mitogen-Activated Protein Kinases; Protein Kinase Inhibitors; Pyridines

Topics: Adult; Child; Colitis, Ulcerative; Crohn Disease; Humans; Interleukin-8

Topics: Adult; Colitis, Ulcerative; Escherichia coli Infections; Female; Humans; Interleukin-8; Pyoderma Gangrenosum

To elucidate the molecular mechanisms involved in the therapeutic effects of granulocyte/monocyte adsorption apheresis, changes were investigated in the cytokine responses of peripheral blood mononuclear cells (PBMC) before and after granulocyte/monocyte adsorptive apheresis in ulcerative colitis (UC) patients. Four patients with active UC were enrolled. All patients responded to granulocyte/monocyte adsorptive apheresis. A total of 20 sessions of four patients were analyzed. Peripheral blood mononuclear cells were isolated from peripheral venous blood within 5 min before and after each session of granulocyte/monocyte adsorptive apheresis. The cells were stimulated with interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha for 24 h, and the secreted IL-8 and IL-6 levels were determined by enzyme-linked immunosorbent assay (ELISA). IL-1beta-induced IL-8 and IL-6 secretion was significantly decreased after granulocyte/monocyte adsorptive apheresis. TNF-alpha-induced IL-8 secretion was also significantly decreased after apheresis, but there was no significant difference in TNF-alpha-induced IL-6 secretion (P = 0.052). In conclusion, granulocyte/monocyte adsorptive apheresis down-regulates the IL-1beta- and TNF-alpha-induced inflammatory responses in PBMC. The induction of hyporesponsiveness to pro-inflammatory cytokines may be an important factor mediating the clinical effects of granulocyte/macrophage adsorptive apheresis in UC patients.

Topics: Adult; Blood Component Removal; Colitis, Ulcerative; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Granulocytes; Humans; Inflammation Mediators; Inflammatory Bowel Diseases; Interleukin-1; Interleukin-6; Interleukin-8; Male; Middle Aged; Monocytes; Treatment Outcome; Tumor Necrosis Factor-alpha

Recognition of repeat CpG motifs, which are common in bacterial, but not in mammalian, DNA, through Toll-like receptor (TLR)9 is an integral part of the innate immune system. As the role of TLR9 in the human gut is unknown, we determined the spectrum of TLR9 expression in normal and inflamed colon and examined how epithelial cells respond to specific TLR9 ligand stimulation. TLR9 expression was measured in human colonic mucosal biopsies, freshly isolated human colonic epithelial cells and HT-29 cells by reverse transcriptase-polymerase chain reaction or Western blotting. Colonic epithelial cell cultures were stimulated with a synthetic CpG-oligodeoxynucleotide (ODN), exhibiting strong immunostimulatory effects in B cells. Interleukin (IL)-8 secretion was determined by enzyme-linked immunosorbent assay, nuclear factor-kappaB (NF-kB) activity by electrophoretic mobility shift assay and IkB phosphorylation by Western blotting. TLR9 mRNA was equally expressed in colonic mucosa from controls (n = 6) and patients with ulcerative colitis or Crohn's disease disease (n = 13). HT-29 cells expressed TLR9 mRNA and protein and responded to CpG-ODN (P < 0.01), but not to non-CpG-ODN stimulation, by secreting IL-8, apparently in the absence of NF-kB activation. Primary epithelial cells isolated from normal human colon expressed TLR9 mRNA, but were completely unresponsive to CpG-ODN stimulation in vitro. In conclusion, differentiated human colonic epithelial cells are unresponsive to TLR9 ligand stimulation in vitro despite spontaneous TLR9 gene expression. This suggests that the human epithelium is able to avoid inappropriate immune responses to luminal bacterial products through modulation of the TLR9 pathway.

Topics: Adult; Aged; Aged, 80 and over; Blotting, Western; Cell Line, Transformed; Cells, Cultured; Colitis, Ulcerative; Colon; Crohn Disease; Epithelial Cells; Female; Gene Expression Regulation; Humans; I-kappa B Proteins; Interleukin-8; Intestinal Mucosa; Male; Membrane Glycoproteins; Middle Aged; NF-kappa B; Oligodeoxyribonucleotides; Receptors, Cell Surface; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Toll-Like Receptor 9; Toll-Like Receptors

Pouchitis is the major long-term complication after ileal pouch-anal anastomosis (IPAA) in patients operated on for ulcerative colitis. The cause is unknown, but both the history of ulcerative colitis and increased bacterial concentration are important factors. Chemokines are mediators for the recruitment of inflammatory cells to the site of inflammation. This study examined the tissue expression of a panel of specific chemokines and the corresponding recruitment of inflammatory cells in IPAA tissue with and without inflammation and after antibiotic treatment.. Biopsy specimens postoperatively from ulcerative colitis patients with IPAA were obtained by endoscopy. Biopsies were taken from 8 patients with noninflamed IPAA and from 14 patients with an episode of acute pouchitis, before and after antibiotic treatment. Biopsies were stained for CD68, CD3, elastase, eotaxin, IP-10, MCP-1, MCP-3, and IL-8 and analyzed by NIH Image analyzer.. Expression of IL-8, MCP-1, MCP-3, and IP-10 was significantly higher in pouchitis than normal pouch. The expression of MCP-1, MCP-3 and IP-10 were significantly lower after antibiotic treatment.. These data support the importance of chemokines for the leukocyte recruitment in pouch tissue during acute pouchitis.

Topics: Adult; Antineoplastic Agents; Chemokine CCL2; Chemokine CXCL10; Chemokines, CC; Chemokines, CXC; Ciprofloxacin; Colitis, Ulcerative; Drug Therapy, Combination; Female; Humans; Immunohistochemistry; Interleukin-8; Male; Pouchitis; Proctocolectomy, Restorative; Rifamycins; Rifaximin

Oral aloe vera gel is widely used by patients with inflammatory bowel disease and is under therapeutic evaluation for this condition.. To assess the effects of aloe vera in vitro on the production of reactive oxygen metabolites, eicosanoids and interleukin-8, all of which may be pathogenic in inflammatory bowel disease.. The anti-oxidant activity of aloe vera was assessed in two cell-free, radical-generating systems and by the chemiluminescence of incubated colorectal mucosal biopsies. Eicosanoid production by biopsies and interleukin-8 release by CaCo2 epithelial cells in the presence of aloe vera were measured by enzyme-linked immunosorbent assay.. Aloe vera gel had a dose-dependent inhibitory effect on reactive oxygen metabolite production; 50% inhibition occurred at 1 in 1000 dilution in the phycoerythrin assay and at 1 in 10-50 dilution with biopsies. Aloe vera inhibited the production of prostaglandin E2 by 30% at 1 in 50 dilution (P = 0.03), but had no effect on thromboxane B2 production. The release of interleukin-8 by CaCo2 cells fell by 20% (P < 0.05) with aloe vera diluted at 1 in 100, but not at 1 in 10 or 1 in 1000 dilutions.. The anti-inflammatory actions of aloe vera gel in vitro provide support for the proposal that it may have a therapeutic effect in inflammatory bowel disease.

Topics: Adult; Aged; Aloe; Anti-Inflammatory Agents; Caco-2 Cells; Colitis, Ulcerative; Eicosanoids; Female; Gels; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Phytotherapy; Plant Extracts; Reactive Oxygen Species

The aim of the study was to assess receptor CD11b and CD62L expression on neutrophils and IL-1beta, IL-6 and IL-8 level in children with ulcerative colitis. Thirty children aged 8-17 years were included in the study. The investigations were performed in the acute phase, prior to the treatment and during remission, 2 months after clinical improvement was obtained. The control group consisted of 12 children with chronic obstipation. IL-1beta, IL-6 and IL-8 serum level and receptor CD11b expression, especially on resting neutrophils, in children with severe and moderate course of the disease were statistically significantly higher; whereas CD62L expression was significantly lower in comparison with the controls and got back to normal during remission. Increased IL-1beta level was observed only in children with severe disease course. In children with mild process the results of the investigations were similar to the control group. In children with severe disease activity the proinflammatory cytokines levels in serum were elevated. The neutrophils in children with severe course of the disease manifested priming in peripheral blood before leaving the circulation.

Topics: Adolescent; CD11b Antigen; Child; Colitis, Ulcerative; Cytokines; Female; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Male; Neutrophils

Mucosal changes in inflammatory bowel disease (IBD) are characterized by ulcerative lesions accompanied by prominent cellular infiltrates in the bowel wall. Chemokines are chemotactic cytokines that are able to promote leukocyte migration to areas of inflammation and are also able to initiate cell activation events. They have recently been implicated in the pathophysiology of many disease states. The aim of this study was to detail the degree and distribution of specific chemokines, interleukin (IL)-8, monocyte chemoattractant protein (MCP)-1, -2, and -3, and macrophage inflammatory protein (MIP)-1alpha and -1beta, in IBD mucosa. Thirty-nine patients were included, ten controls, 20 ulcerative colitis (UC), and nine Crohn's disease (CD), with a range of disease activity. Colonic mucosal biopsies were collected from UC, CD, and control patients and embedded in glycol methacrylate. Two-micrometre-thick sections were cut and stained using immunohistochemistry for chemokine protein expression. Sections were analysed using a light microscope. Expression of all types of chemokine protein was detected in colonic mucosa from both control and IBD patients. Patterns of staining between IBD patients and controls differed significantly, but CD and UC patients demonstrated similar patterns of staining. Individual chemokine expression was found to be significantly up-regulated in IBD when patients were compared with the non-diseased group in all areas of the mucosal sections. Up-regulated chemokine expression correlated with increasing activity of the disease. It is concluded that human colonic chemokine expression is non-selectively up-regulated in IBD. The results supported the hypothesis that the degree of local inflammation and tissue damage in UC and CD is dependent on local expression of specific chemokines within IBD tissues.

Topics: Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL7; Chemokine CCL8; Chemokines; Colitis, Ulcerative; Colon; Crohn Disease; Cytokines; Humans; Immunohistochemistry; Interleukin-8; Intestinal Mucosa; Macrophage Inflammatory Proteins; Monocyte Chemoattractant Proteins; Up-Regulation

In the present study we have used RT-PCR to investigate nicotinic acetylcholine receptor (nAChR) subunit expression, and studied the effect of nicotine on TNFalpha-induced cytokine (IL-8) release in the epithelial cell line HT29. RNA was extracted using a commercial kit and amplified by RT-PCR. RT-PCR products were separated by electrophoresis and visualised using ethidium bromide. IL-8 release was measured by ELISA from cells activated for 6 h with TNFalpha (50 ng ml(-1)) in the absence and presence of nicotine (10(-11)-10(-6) M). HT29 cells contained mRNA for beta1, alpha4, alpha5, and alpha7 nAChR subunits. Activation of HT29 cells increased IL-8 release from undetectable amounts to 3.92 +/- 0.51 ng ml(-1) (n = 5). Nicotine significantly inhibited TNFalpha-induced IL-8 release in a concentration related manner with peak inhibition occurring at 10(-7) M (2.39 +/- 0.78 ng ml(-1), n = 5). Our data suggests that, while HT29 cells express mRNA for nAChR subunits, the only nAChR subunits that could form functional receptors and inhibit IL-8 release are alpha7.

Topics: Colitis, Ulcerative; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; HT29 Cells; Humans; Interleukin-8; Nicotine; Nicotinic Agonists; Receptors, Nicotinic; Reverse Transcriptase Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

To investigate if nuclear factor-kappa B (NF-kappa B) p65 antisense oligonucleotides might affect the expression of NF-kappa B p65 and cytokines in lamina propria mononuclear cells(LPMC) from patients with ulcerative colitis (UC). LPMC were isolated from intestinal mucosal biopsy specimens from 3 patients with UC, and cultured with or without NF-kappa B p65 antisense oligonucleotides (5'-GGAACAGTTCGTCCTATGG-3'), missense oligonucleotides (5'-GGAACAGTTCGTCTATGG-3') and dexamethasone. NF-kappa B p65 expression was determined by western blot analysis. The expression of cytokine mRNA was studied by reversal transcription-polymerase chain reaction (RT-PCR). The cytokine levels were measured by enzyme linked immunosorbent assay. The results showed that NF-kappa B p65 antisense oligonucleotides resulted in down-regulation of NF-kappa B p65 expression, blocked the expression of IL-1 beta mRNA and IL-8 mRNA, and strikingly reduced the production of IL-1 beta and IL-8, and these effects were greater than those of dexamethasone in cultured LPMC from patients with UC(P < 0.05). Therefore, the application of NF-kappa B p65 antisense oligonucleotides may serve as a novel molecular approach for the treatment of patients with UC.

Topics: Cells, Cultured; Colitis, Ulcerative; Cytokines; Humans; Interleukin-1; Interleukin-8; Intestinal Mucosa; Monocytes; NF-kappa B; Oligonucleotides, Antisense; RNA, Messenger

The level of macrophage migration inhibitory factor (MIF) and the functions of dendritic cells (DC) are up-regulated in the peripheral blood, and the numbers of MIF-expressing cells and mature DC are increased at the colonic mucosa from patients with ulcerative colitis (UC). However, a functional relationship between MIF and DC, and the role of MIF in the pathogenesis of UC, are not clear. In this study, we showed that a pure population of peripheral blood DC is a new and still unknown source of MIF. DC from UC patients produced significantly higher levels of MIF (17 x 5 +/- 9 x 8 ng/ml, n = 10) compared with patients with Crohns disease (CD) (4 x 6 +/- 2 x 5 ng/ml, n = 5, P< 0 x 01) and control subjects (5 x 0 +/- 2 x 6 ng/ml, n = 10, P< 0 x 01). A double immunofluorescence study revealed the expression of MIF by CD83-positive mature DC at the colonic mucosa from UC patients. Blood DC treated with high amounts of MIF (500 ng/ml) showed a significantly higher stimulatory capacity (43287 +/- 5998 CPM, n = 5) in an allogenic mixed leucocyte reaction compared with untreated DC (27528 +/- 8823 CPM, n = 5, P< 0 x 05). Study of intracellular cytokine expression showed that MIF induced significant levels of interleukin (IL)-1 and IL-8 in monocytes and DC from UC and CD patients. These results showing the capacity of MIF to induce increased functional capacity of DC, and to produce IL-1 and IL-8 from monocytes and DC, indicate a role of MIF in the induction and/or perpetuation of the inflammatory environment in UC.

Topics: Cells, Cultured; Colitis, Ulcerative; Colon; Crohn Disease; Dendritic Cells; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Intestinal Mucosa; Macrophage Migration-Inhibitory Factors; Monocytes; Tumor Necrosis Factor-alpha

To investigate the activation of nuclear factor-kappaB (NF-kappaB) and its relationship with expression of cytokine mRNA in intestinal mucosal biopsy specimens from patients with ulcerative colitis (UC).. 31 cases with UC were included in the study. 17 cases received sulfasalazine (SASP) or SASP and glucocorticoid treatment. 14 cases did not receive any medication related with UC. Normal mucosa from 11 colon cancer cases served as control. Ten pieces of intestinal mucosal biopsy specimens were obtained from each patient. NF-kappaB DNA binding activity was evaluated with electrophoretic mobility shift assay (EMSA). Expression of cytokine mRNA were studied with reversal tanscription-polymerase chain reaction (RT-PCR).. (1) The expression of IL-1beta mRNA and IL-8 mRNA was increased significantly in patients with UC, as compared with that in the control specimens (P < 0.05) and had a significant positive correlation with NF-kappaB DNA binding activity (r = 0.8363, P < 0.05; r = 0.6024, P < 0.05, respectively). (2) Glucocorticoids and SASP strongly inhibited NF-kappaB activation and signficantly decreased the expression of IL-1beta mRNA and IL-8 mRNA.. NF-kappaB is a major and essential factor in regulating the expression of cytokine and plays a fundamental role in the pathogenesis of UC. SASP and glucocorticoids decrease cytokine expression via inhibition of NF-kappaB activation.

Topics: Adult; Colitis, Ulcerative; Colon; Cytokines; Drug Therapy, Combination; Female; Glucocorticoids; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Mucous Membrane; NF-kappa B; RNA, Messenger; Sulfasalazine

The local release of human neutrophil lipocalin, considered to be highly specific for neutrophil granulocyte activation, and interleukin-8 and tumor necrosis factor-alpha were studied in 11 patients with distal ulcerative colitis and proctitis before and during treatment with steroid enemas. A rectal perfusion technique for sampling and specific immunoassays for analysis were used. In responders (N = 8) the concentrations of all proteins decreased during the study. There was a close correlation between human neutrophil lipocalin concentrations and treatment response. Tumor necrosis factor-alpha showed an initial decline in concentrations irrespective of treatment outcome and preceded the decline of human neutrophil lipocalin and interleukin-8. We conclude that decreased neutrophil degranulation is correlated with treatment outcome. Furthermore, an important role of tumor necrosis factor-alpha in the process of stimulating neutrophil activation and degranulation in ulcerative colitis is suggested.

Topics: Acute-Phase Proteins; Anti-Inflammatory Agents; Carrier Proteins; Colitis, Ulcerative; Enema; Humans; Interleukin-8; Lipocalin-2; Lipocalins; Neutrophils; Oncogene Proteins; Prednisolone; Proctitis; Proto-Oncogene Proteins; Tumor Necrosis Factor-alpha

The aim of this study was to clarify the correlation between cytokine profile in colonic mucosa with disease activity and response to granulocytapheresis (GCAP) in patients with ulcerative colitis (UC), using a reliable, reproducible quantitative method.. Colonoscopic biopsies of inflamed colonic mucosa (16 patients, 21 cases) and uninflamed colonic mucosa (25 patients, 33 cases) were obtained from UC patients. Messenger (m)RNA was extracted and subjected to realtime polymerase chain reaction for quantitative measurement of interleukin (IL)-12, interferon-gamma, tumor necrosis factor-alpha, IL-4, IL-8, and IL-18 mRNAs. In seven patients with high disease activity despite prednisolone (PSL) treatment (> or = 20 mg/day), one course of GCAP was conducted, and pre- and post-GCAP cytokine profiles were determined.. In inflamed colonic mucosa of UC patients, three cytokine profiles were observed: 1) high expression of interferon-gamma, tumor necrosis factor-alpha, and IL-4 mRNAs but low expression of IL-8 mRNA; 2) high expression of IL-8 mRNA and low expression of others; and 3) low expression of all cytokines examined. Inflamed colonic mucosa of patients with high disease activity showed the second pattern. Inflamed colonic mucosa of patients who were not treated with PSL and who had low disease activity showed the first pattern, whereas those on high-dose PSL exhibited the second pattern. IL-8 mRNA was significantly higher in inflamed UC samples than in uninflamed samples. GCAP was effective in five of seven PSL-resistant patients (71.4%). IL-8 was the only cytokine that correlated with effectiveness of GCAP. Compared with GCAP nonresponders, responders had significantly higher IL-8 mRNA before GCAP and showed marked reduction of IL-8 mRNA after GCAP.. IL-8 mRNA was significantly increased in inflamed mucosa of UC. Patients with high IL-8 mRNA expression in colonic mucosa despite PSL treatment were responsive to GCAP. Therefore, quantitative measurement of mucosal IL-8 mRNA may be useful in predicting the response to GCAP.

Topics: Acute Disease; Adult; Aged; Chronic Disease; Colitis, Ulcerative; Colon; Cytokines; DNA, Complementary; Female; Granulocytes; Humans; Inflammation; Interferon-gamma; Interleukin-4; Interleukin-8; Intestinal Mucosa; Leukapheresis; Male; Middle Aged; Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha

Lidocaine and related local anaesthetics have been shown to be effective in the treatment of ulcerative colitis (UC). However, the mechanisms underlying their therapeutic effect are poorly defined. Intestinal epithelial cells play an important role in the mucosal inflammatory response that leads to tissue damage in UC via the secretion of pro-inflammatory cytokines and chemokines. The aim of this study was to evaluate the direct immunoregulatory effect of lidocaine on pro-inflammatory cytokine and chemokine secretion from intestinal epithelial cells. HT-29 and Caco-2 cell lines were used as a model system and treated with lidocaine and related drugs. The expression of IL-8, IL-1beta and the IL-1 receptor antagonist (RA) were assessed by ELISA and quantification of mRNA. In further experiments, the effect of lidocaine on the secretion of IL-8 from freshly isolated epithelial cells stimulated with TNFalpha was tested. Lidocaine, in therapeutic concentrations, inhibited the spontaneous and TNFalpha-stimulated secretion of IL-8 and IL-1beta from HT-29 and Caco-2 cell lines in a dose-dependent manner. Similarly, suppression of IL-8 secretion was noted in the freshly isolated epithelial cells. Other local anaesthetics, bupivacaine and amethocaine, had comparable effects. Lidocaine stimulated the secretion of the anti-inflammatory molecule IL-1 RA. Both the inhibitory and the stimulatory effects of lidocaine involved regulation of transcription. The results imply that the therapeutic effect of lidocaine may be mediated, at least in part, by its direct effects on epithelial cells to inhibit the secretion of proinflammatory molecules on one hand while triggering the secretion of anti-inflammatory mediators on the other.

Topics: Adenocarcinoma; Anesthetics, Local; Bupivacaine; Colitis, Ulcerative; Colonic Neoplasms; Epithelial Cells; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-8; Intestinal Mucosa; Lidocaine; RNA, Messenger; Sialoglycoproteins; Tetracaine; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Accumulation and infiltration by neutrophil granulocytes is a prominent feature in the local inflammatory process in ulcerative colitis (UC). The present study was performed to evaluate human neutrophil lipocalin (HNL) as a specific neutrophil marker in the inflamed lesions of the colon and rectum in patients with colitis and proctitis.. The activity of intestinal neutrophils with respect to release of granule proteins was studied in 18 patients with UC (10 with colitis and eight with isolated proctitis) and in 18 healthy controls using perfusion fluid and biopsies from the sigmoid colon and rectum. The released amounts of the neutrophil granule proteins HNL and myeloperoxidase (MPO) were determined by radioimmunoassays, and the location of HNL and MPO in biopsies from colonic mucosa was examined by immunohistochemistry.. Mucosal release of HNL and MPO was increased 10-55-fold in patients with colitis and proctitis compared with controls. Their bowel biopsies demonstrated that only neutrophils were stained with anti-HNL. We also found correlations between HNL and levels of granulocyte/macrophage-colony stimulating factor (GM-CSF) and interleukin 8 (IL-8) in perfusion fluids from the sigmoidal segments of patients with proctitis, between HNL and GM-CSF in rectal segments in patients with proctitis, and in sigmoidal segments in patients with colitis.. We conclude that the increased release of HNL and MPO in colorectal perfusion fluids indicates neutrophil involvement in the local inflammatory process, and suggest that HNL may serve as a specific marker of intestinal neutrophil activation in UC. GM-CSF, and to some extent IL-8, may play a role in neutrophil accumulation and priming in this disease.

Topics: Acute-Phase Proteins; Adult; Aged; Biomarkers; Carrier Proteins; Colitis, Ulcerative; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunohistochemistry; Interleukin-8; Intestinal Mucosa; Lipocalin-2; Lipocalins; Male; Middle Aged; Neutrophils; Oncogene Proteins; Peroxidase; Proctitis; Proto-Oncogene Proteins

Clinical differences between small- and large-bowel Crohn's disease have been demonstrated. Neutrophil migration and degranulation are important effector mechanisms in gut damage. Granulocyte elastase, a neutrophil-bound enzyme, interleukin 8 and 1beta can be detected in whole-gut lavage fluid. We aimed to assess differences between large- and small-bowel Crohn's disease.. A total of 167 patients with active inflammatory bowel disease (118 Crohn's disease, 49 ulcerative colitis) underwent whole-gut lavage with a polyethylene glycol electrolyte solution. Granulocyte elastase was assayed using an enzyme substrate reaction, IL-8 and IL-1beta by ELISA.. Twenty-seven of 36 patients with isolated colonic Crohn's disease had detectable granulocyte elastase (median 0.259 pKat/l, range < 0.039-2.742 microKat/l), whereas 3 of 15 with small-bowel involvement alone had detectable granulocyte elastase (median < 0.039 microKat/l, range < 0.039-0.266 microKat/l; P < 0.0001). Granulocyte elastase levels were significantly higher in patients with ileocolonic disease and post-ileocaecal resection compared with small-bowel disease alone. IL-8 (P< 0.0001) and IL-1beta (P < 0.04) levels differed between colonic and ileal distributions. No variations were seen in ulcerative colitis.. Neutrophil migration to the gut lumen in Crohn's disease is a feature of colonic disease irrespective of associated ileal lesions. This suggests that bacterial-derived chemo-attractants may play a role. High levels of IL-8 in colonic disease are consistent with this hypothesis.

Topics: Adult; Chemotaxis, Leukocyte; Colitis, Ulcerative; Crohn Disease; Female; Humans; Interleukin-1; Interleukin-8; Intestine, Large; Intestine, Small; Leukocyte Elastase; Male; Middle Aged; Neutrophils; Therapeutic Irrigation

This study aimed to determine if mucosal expression of the chemokines IL-8, RANTES and MCP-1 and the pro-inflammatory cytokines TNFalpha and IL-6 are elevated in patients with inflammatory bowel disease.. Intestinal mucosa samples were obtained at the time of surgical resection, n = 16 from each of the following groups: normal/control, CD and UC.. An homogenate was prepared of each tissue sample and cytokines measured by ELISA.. IL-8 was significantly increased in both disease groups compared to controls Similarly, RANTES levels were also significantly increased. MCP-1 levels were increased in both disease groups, this increase was statistically significant in the UC group only. TNFalpha and IL-6 were significantly increased in the CD group only.. Chemokines, together with key cytokines that promote their release are elevated in mucosal tissues from patients with IBD. It is likely that these chemokines play an important role in the perpetuation of tissue destructive inflammatory processes.

Topics: Chemokine CCL2; Chemokine CCL5; Chemokines; Colitis, Ulcerative; Colon; Crohn Disease; Cytokines; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-8; Intestinal Mucosa

Delayed neutrophil apoptosis is a feature of persistent acute inflammation. Neutrophil-mediated damage has been shown to be associated with the development of inflammatory bowel disease (IBD). Persistence of these cells both at the colonic site and circulation may further contribute to IBD. The aims of this study were to determine whether neutrophils isolated from IBD patients delay apoptosis and to investigate possible mechanisms involved in this delay. We studied 20 patients with IBD, 13 with Crohn's disease, and 7 with ulcerative colitis, all of whom were undergoing intestinal resection for symptomatic disease. Seventeen patients undergoing elective resection of colon cancer acted as operative controls. Systemic, mesenteric arterial, and mesenteric venous blood was harvested. Neutrophils isolated from patients with IBD showed decreased spontaneous apoptosis compared to cancer patients. Mesenteric venous serum of IBD patients contributed to this delay, which contained higher concentrations of interleukin-8 (IL-8). Pro-caspase 3 expression was also reduced in IBD neutrophils, which may contribute to decreased spontaneous and Fas antibody-induced apoptosis. Neutrophil apoptosis may be altered in Crohn's disease and ulcerative colitis through release of anti-apoptotic cytokines and altered caspase expression. The alterations in cell death mechanisms may lead to persistence of the inflammatory response associated with IBD.

Topics: Adolescent; Adult; Aged; Apoptosis; Case-Control Studies; Caspase 8; Caspase 9; Caspases; Colitis, Ulcerative; Crohn Disease; Enzyme Precursors; Female; Humans; Hydrocortisone; In Vitro Techniques; Interleukin-8; Male; Middle Aged; Neutrophils

Interleukin-8 (IL-8) as an alpha-chemokine recruits and activates neutrophils, which are abundant in the intestinal lesions of ulcerative colitis (UC) and Crohn's disease (CD). Stromal cell-derived factor 1 (SDF1-alpha) is a new chemokine that is chemotactic to neutrophils. The aims of this study were to assess the relative expression of SDF1-alpha and IL-8 mRNA in different colonic regions and patients with inflammatory bowel disease with varied degrees of inflammation in the colon.. Colon biopsy samples were obtained from 19 patients with UC, 12 with CD, and 5 with irritable bowel syndrome (IBS) who underwent colonoscopy. Levels of IL-8 and SDF1-alpha mRNA expression were measured semiquantitatively by reverse-transcription and polymerase chain reaction amplification. The cytokine mRNA levels were corrected for glyceraldelyde-3-phosphate dehydrogenase mRNA expression.. IL-8 mRNA expression was significantly correlated with SDF1-alpha expression in normal biopsies from IBS patients (r = 0.58, p < 0.01). There was no significant difference in cytokine mRNA expression (IL-8 or SDF1-alpha) across different regions of the colon or rectum in uninflamed normal biopsies. The IL-8 mRNA expression ratios in UC (mean +/- SD, 1.03 +/- 0.52) and CD (0.90 +/- 0.38) patients were significantly higher than in IBS (0.52 +/- 0.17) (p < 0.01, p < 0.05, respectively). The SDF1-alpha mRNA expression ratio in UC (0.30 +/- 0.52) was higher than in both CD (0.21 +/- 0.10) and IBS patients (0.22 +/- 0.11) (p < 0.01, <0.05, respectively). A statistically significant correlation was found between the IL-8 mRNA expression and the colonic inflammation in UC patients (r = 0.44, p < 0.05) but not for SDF1-alpha expression in UC patients.. IL-8 but not SDF1-alpha mRNA expression was associated with inflammation in UC. This suggests that IL-8 may play a more important role in inflammatory bowel disease than does SDF1-alpha.

Topics: Adult; Biopsy; Case-Control Studies; Chemokine CXCL12; Chemokines, CXC; Colitis, Ulcerative; Colon; Colonic Diseases, Functional; Crohn Disease; Female; Glyceraldehyde-3-Phosphate Dehydrogenases; Humans; Interleukin-8; Male; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stromal Cells

We previously reported that intracolonic administration of enprostil, a prostaglandin-E(2) (PGE(2)) analogue, had therapeutic effects on acute colitis induced in rodents by dextran sulfate sodium (DSS). In addition, production of growth-regulated gene product/cytokine-induced neutrophil chemoattractant-1 [GRO/CINC-1; an interleukin(IL)-8 like cytokine] was suppressed in the inflamed tissues. In the present study we used a human colon cancer cell line (HT-29) to investigate enprostil effects on the IL-8 production of intestinal epithelial cells stimulated by various stimulants. In a MTT assay, concentrations of enprostil >10(-5)M had cytotoxitic effects on HT-29 cells. Furthermore, 10(-6) M enprostil suppressed IL-8 production in HT-29 cells, SW620 and CaCo2 stimulated with interleukin-1 beta (IL-1 beta) or lipopolysaccharide (LPS), but did not suppress this response when cells were stimulated with tumour necrosis factor (TNF)-alpha. These results suggest that enprostil affects a point in the pathway between the IL-1 receptor or LPS receptor and nuclear factor-kappa B(NF-kappa B), without affecting the pathway between the TNF receptor and NF-kappa B, with the latter factor being required for the IL-8 gene transcription. The therapeutic effect of exogenous enprostil on DSS colitis may involve the inhibition of IL-8 production in colonic epithelial cells stimulated by IL-1 beta or LPS.

Topics: Colitis, Ulcerative; Colon; Cyclic AMP; Dinoprostone; Enprostil; Humans; Interleukin-1; Interleukin-8; Intestinal Mucosa; Lipopolysaccharides; RNA, Messenger; RNA, Neoplasm; Signal Transduction; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

To evaluate the monitoring action of the expression of IL-1 beta, IL-8 mRNA, myeloperoxidase (MPO) and superoxide dismutase (SOD) activity in the active ulcerative celitis (UC).. Twenty active UC patients, 23 inactive UC patients; and 14 non-UC inflammation patients were selected. Twelve patients who complained of flatulence, abdominal pain and constipation receiving endoscopic examination were used as control. MPO and SOD activity, IL-1 beta and IL-8 mRNA expression (hybridization in situ) were determined on the endoscopic biopsy specimens of all patients. In 20 active UC patients, 14 patients received endoscopic examination and 4 indices detection were done once again after 2 months' treatment with prednisone and SASP.. Mucosal MPO activities of active UC patients, inactive UC patients, and non-UC inflammation patients are higher than those of control patients, (19.37 +/- 0.54, 11.59 +/- 1.41, and 12.97 +/- 0.49) U/g tissue vs (9.49 +/- 0.51) U/g tissue (P < 0.01). SOD activities are lower than that of control patients, (5.03 +/- 07,7. 7.66 +/- 0.79, and 6.98 +/- 0.61) U/mg protein vs (8.82 +/- 0.58) U/mg protein (P < 0.05). Mucosal MPO activity of active UC patients is also higher than that of inactive and non-UC inflammation patients (P < 0.01); while SOD activity is lower than them (P < 0.01). After 2 months' medical treatment, MPO activity of 14 active UC patients decreased, (12.61 +/- 0.74) U/g tissue vs (19.31 +/- 0.44) U/g tissue (P < 0.01), while SOD activity elevated (7.44 +/- 0.55) U/mg protein vs (5.10 +/- 1.05) U/mg protein (P < 0.05), compared with that of before treatment. Positive expression of IL-1 beta mRNA appeared in the epithelial and inflammatory cells of all active UC patients, 9 inactive UC patients, and 7 non-UC inflammation patients. While expression of IL-8 mRNA only appeared positively in all active UC patients. In 14 active UC patients, there were no detection of interleukin mRNA expression after 2 months' treatment.. Mucosal MPO, SOD, IL-1 beta, and IL-8 mRNA could be used as 4 indices monitoring the activity of UC. And IL-1 beta mRNA is also helpful to supervise early or late UC activity.

Topics: Biomarkers; Colitis, Ulcerative; Humans; Interleukin-1; Interleukin-8; Peroxidase; RNA, Messenger; Superoxide Dismutase

Infiltration of neutrophils and their release of toxic reactive oxygen species (ROS) in the colonic mucosa are associated with tissue damage in ulcerative colitis (UC). This neutrophil migration may be induced by chemoattractants, such as cytokines, in the colonic milieu. One such chemoattractant is interleukin-8 (IL-8), a neutrophil chemokine that is present at high concentrations in inflamed mucosa. However, the functional significance of IL-8 in neutrophil attraction and activation in UC has not been established. We hypothesized that IL-8 in the colonic lumen of patients with UC primes neutrophils, leading to their attraction and activation.. The colonic milieu was sampled by rectal dialysis. Using a semi-permeable membrane with a molecular weight cut-off of 12 kDa, dialysis solution was placed in the rectum and allowed to equilibrate over a 4-h period with the colonic milieu of controls or of patients with UC. IL-8 concentrations were measured by ELISA. Two functions of healthy neutrophils (PMN) were measured: expression of CD11-b surface adhesion molecules (by flow cytometry), and production of ROS (by both chemiluminescence and cytochrome C reduction assays). Neutrophil functions after exposure to rectal dialysates or n-formyl-methionyl-leucyl-phenylalanine (fMLP) were assessed before and after adding anti-IL-8 antibody or the fMLP blocker BMLP.. IL-8 concentrations in dialysates from patients with active UC were significantly higher than in controls and correlated with disease activity. UC dialysates significantly increased ROS production and CD11-b expression by neutrophils and anti-IL-8 antibody partially (50%) inhibited these stimulatory effects of UC dialysates. Preincubation of neutrophils with UC dialysates significantly potentiated the fMLP-induced rise in ROS and anti-IL-8 antibody completely abolished this priming effect.. The colonic milieu, sampled by rectal dialysis, from patients with active UC can both activate and prime neutrophils in vitro. High concentrations of IL-8 in the colonic lumen of UC patients are partially responsible for the activating effects of rectal dialysates, and account for all of its priming effects. These findings provide direct evidence for a role for IL-8 in inflammatory bowel disease.

Topics: Adult; Colitis, Ulcerative; Colon; Dialysis Solutions; Flow Cytometry; Humans; Interleukin-8; Luminescent Measurements; Macrophage-1 Antigen; Middle Aged; Neutrophil Activation; Neutrophils; Reactive Oxygen Species; Rectum

Proinflammatory cytokines are believed to be involved in the pathogenesis of ulcerative colitis (UC). The aim of this study was to clarify the profiles of proinflammatory cytokine production in patients with UC in terms of disease intractability, endoscopic findings, and host response to lipopolysaccharide (LPS) stimulation. Colonic mucosal tissues were obtained from patients with active UC (n = 15, including 4 patients with intractable disease) and inactive UC (n = 7), non-inflammatory bowel disease (IBD) colitis (n = 11), and controls (n = 20). Organ culture was performed, and the amounts of four cytokines (described below) in the culture media were determined by enzyme-linked immunosorbent assay (ELISA). LPS stimulation enhanced interleukin (IL)-1beta, IL-8, and IL-6 production in colonic specimens from all groups, but enhanced tumor necrosis factor (TNF)-alpha production only in active UC specimens. Levels of IL-6, IL-8, and TNF-alpha were significantly higher in active UC than in non-IBD colitis, and the production of all three of these cytokines was correlated to the endoscopic grade of inflammation. The production of these cytokines was also significantly higher in patients with intractable disease receiving corticosteroids than in patients with non-intractable disease receiving corticosteroids. These results suggest that enhanced production of mucosal proinflammatory cytokines may be implicated in the pathogenesis of UC.

Topics: Adult; Aged; Biomarkers; Colitis, Ulcerative; Colonoscopy; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Glucocorticoids; Humans; Immunity, Mucosal; Interleukin-1; Interleukin-6; Interleukin-8; Intestinal Mucosa; Lipopolysaccharides; Male; Middle Aged; Organ Culture Techniques; Tumor Necrosis Factor-alpha

Anecdotal reports suggest that smoking may be beneficial for patients with inflammatory bowel disease (IBD) as nicotine may act through inflammatory mediators within the colonic mucosa. Furthermore, there is increasing evidence that cytokines play a pathologic role in IBD. Our aim was to determine the effects of cigarette smoking on cytokine levels in the colonic mucosa of patients with and without IBD. Mucosal biopsies were obtained from 10 patients with Crohn's disease (CD), 10 with ulcerative colitis (UC), and 10 healthy controls. Five of 10 patients in each of the three groups were smokers and five were nonsmokers. Concentrations of interleukin (IL)-1beta, IL-2, IL-6, and IL-8 were determined using enzyme-linked immunosorbent assay (ELISA). Cytokine levels of smokers were compared with nonsmokers in each group and with controls. Results were analyzed using the Mann-Whitney test; significance was set at p<0.05. The concentration of IL-8 was significantly higher in healthy controls who smoke compared with nonsmokers and significantly reduced in smokers with CD compared with nonsmokers with CD. Moreover, concentrations of IL-1beta and IL-8 were significantly reduced in smokers with UC compared with nonsmokers with UC. Smokers had significantly elevated levels of IL-8 in the colonic mucosa. Smokers with IBD had a significant reduction in cytokine levels; specifically, IL-1beta and IL-8 for patients with UC and IL-8 for patients with CD. Further studies are warranted to determine if this reduction in cytokine levels is histologically and clinically significant.

Topics: Case-Control Studies; Colitis, Ulcerative; Colon; Crohn Disease; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-8; Interleukins; Intestinal Mucosa; Smoking

The role of the eosinophil granulocyte in bowel mucosa in inflammatory bowel disease still remains obscure. The present study was performed in order to elucidate the local eosinophil activity and activating cytokines in the inflamed lesions of colon and rectum in patients with ulcerative colitis and proctitis.. The activity of intestinal eosinophils with respect to the release of granule proteins was studied in 18 patients (10 with colitis and 8 with isolated proctitis) and 18 healthy controls, using intraluminal segmental perfusion of the sigmoid colon and rectum. The released amounts of eosinophil granule proteins: eosinophil cationic protein (ECP), eosinophil peroxidase (EPO), and eosinophil protein X (EPX) to perfusion fluid were determined by radioimmunoassays. The intraluminal release of possible eosinophil priming cytokines granulocyte/macrophage-colony stimulating factor (GM-CSF) and interleukin 8 (IL-8), were analyzed by immunoassays.. The mucosal release of ECP, EPO, and EPX was increased 10- to 20-fold in patients with colitis and proctitis compared with controls. The intraluminal release of GM-CSF and IL-8, was several-fold enhanced in patients with colitis and proctitis. We also found a correlation between all three eosinophil granule proteins and the levels of IL-8/GM-CSF in the sigmoidal segments of patients with colitis.. We conclude that the increased release of ECP, EPO, and EPX to colorectal perfusion fluid indicate eosinophil involvement in the local disease in patients with colitis and proctitis. IL-8 and GM-CSF may play a role in eosinophil accumulation and priming in colitis.

Topics: Adult; Aged; Blood Proteins; Colitis, Ulcerative; Cytokines; Eosinophil Granule Proteins; Eosinophil Peroxidase; Eosinophil-Derived Neurotoxin; Eosinophils; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation Mediators; Interleukin-8; Male; Middle Aged; Peroxidases; Proctitis; Ribonucleases

Chemokines are thought to be important for the recruitment of granulocytes and mononuclear cells and thus for the maintenance of inflammation in ulcerative colitis (UC). We have studied the expression of interferon-gamma inducible protein-10 (IP-10), interleukin-8 (IL-8), monocyte chemoattractant protein (MCP)-1, MCP-3, and macrophage inflammatory protein (MIP)-1alpha in UC patients and control individuals to assess the role of these chemokines in disease progression. Colonic biopsies were taken endoscopically from patients and controls, frozen immediately and subsequently stained for IP-10, IL-8, MCP-1, MCP-3, and MIP-1alpha in serial sections. Cells infiltrating the lamina propria but not epithelial cells express the analyzed chemokines. They were differentiated and counted, and chemokine-expressing cells were quantified by image analysis. The percentage of cells expressing IP-10, IL-8, MCP-1, and MCP-3 was significantly enhanced in all UC samples as compared to controls. Expression in the controls was borderline, except for IP-10. No expression of MIP-1alpha was found in controls and UC. IP-10 was also markedly expressed in the mucosa of control biopsies and therefore could have a role in activated T lymphocytes' recruitment into the healthy mucosa.

Topics: Adult; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL7; Chemokine CXCL10; Colitis, Ulcerative; Colon; Cytokines; Female; Gene Expression; Humans; Immunohistochemistry; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Monocyte Chemoattractant Proteins; Up-Regulation

Neutrophil gelatinase-associated lipocalin (NGAL) is a newly described neutrophil lipocalin that may bind the proinflammatory bacterial tripeptide N-formylmethionyl-leucyl-phenylalanine. In situ hybridization and immunohistochemical studies have shown a strong NGAL expression in colonocytes and neutrophils in ulcerative colitis (UC). Because NGAL is highly protease resistant, it should be ideal for in vivo fecal and dialysate studies. Our aim was to investigate the potential of NGAL as a disease activity marker in UC and to compare it with IL-8 and TNF-alpha.. Twenty-three patients with UC, 14 with Crohn's disease (CD), 19 patients with acute infectious enterocolitis, and 20 healthy controls were included. The disease activity of UC and CD was scored semiquantitatively. Concentrations of NGAL, IL-8, and TNF-alpha were determined in rectal dialysis fluid, feces, and serum using sandwich enzyme-linked immunosorbent assays. The total protein concentration in feces and dialysate fluid was measured, and the amount of markers was expressed as ng/mg protein.. In healthy controls and non-IBD (irritable bowel disease) colitis, the median values for NGAL in feces were 183 ng/mg protein and 546 ng/mg protein (p < 0.01), respectively. When separating UC into clinical activity groups (remission, mild/moderate, and severe disease activity) the corresponding values of NGAL were 442 ng/mg (p > 0.05), 605 ng/mg (p < 0.02), and 3646 ng/mg (p < 0.001, compared with controls), respectively, and in quiescent colonic CD 368 ng/mg (p > 0.05) and in active stages 751 ng/mg (p < 0.01). NGAL levels in dialysis fluid listed in the same order were: 11 ng/mg for controls, 71 ng/mg (p > 0.05) for non-IBD colitis, 100 ng/mg (p < 0.02), 179 ng/mg (p < 0.01), and 2053 ng/mg (p < 0.001) for UC, and 14 ng/mg (p > 0.05) and 121 ng/mg (p < 0.02) for CD, respectively. Serum NGAL concentrations did not differ between UC and CD in quiescent versus active stages. A significant increase of NGAL in both feces and dialysate with increasing disease activity of UC was found (p = 0.02 and p = 0.003, respectively).. The NGAL content in rectal dialysate and particularly in feces seems to be a reliable marker for severe disease activity in UC, whereas serum NGAL concentrations do not reflect disease activity.

Topics: Acute-Phase Proteins; Adult; Aged; Biomarkers; Carrier Proteins; Colitis, Ulcerative; Crohn Disease; Enzyme-Linked Immunosorbent Assay; Feces; Female; Humans; Interleukin-8; Lipocalin-2; Lipocalins; Male; Middle Aged; Neutrophils; Oncogene Proteins; Proto-Oncogene Proteins; Rectum; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8) is a peptide which induces not only chemotaxis of neutrophils but also the release of reactive oxygen metabolites from the neutrophils. There are few reports which clarify the relationships between IL-8 and mucosal infiltration of neutrophils or reactive oxygen metabolites produced by neutrophils in the colonic mucosa of ulcerative colitis (UC). Biopsy specimens of colonic mucosa obtained from 26 patients with active UC and 21 patients with inactive UC were studied in order to clarify the relationships among the inflammation factors in UC. Levels of IL-8 and myeloperoxidase in organ culture media of the biopsy specimens from active UC (measured by ELISA and EIA) were significantly higher than those from inactive UC and controls. Reactive oxygen metabolites of biopsy specimens in active UC (measured by luminol-dependent chemiluminescence) were also markedly increased compared to those in inactive UC and controls. The levels of IL-8 were closely correlated to luminol-dependent chemiluminescence or myeloperoxidase levels. However, the levels of IL-8 and myeloperoxidase did not correlate with the grades of activity on colonoendoscopic findings. These findings suggest that IL-8 may play a role in the pathophysiology of UC but it does not define the endoscopic activity grades of UC.

Topics: Adolescent; Adult; Aged; Cells, Cultured; Colitis, Ulcerative; Enzyme-Linked Immunosorbent Assay; Female; Humans; Indicators and Reagents; Interleukin-8; Intestinal Mucosa; Luminescent Measurements; Luminol; Male; Middle Aged; Neutrophils; Peroxidase

To elucidate the possible role of proinflammatory cytokines in inflammatory bowel disease, the expression and localization of interleukin (IL) -6 and IL-8 mRNAs were examined in colonic biopsy specimens obtained from 10 patients with active ulcerative colitis (UC), 5 with inactive UC, 6 with Crohn's disease (CD), and 5 normal controls. In situ hybridization with digoxigenin-labeled probes and immunohistochemistry for both cytokines were performed. The IL-6 mRNA expression was enhanced in the inflamed mucosa in 4 of 6 CD patients, while that of UC patients stayed at baseline. In contrast, IL-8 mRNA expression was apparently augmented (P = 0.044) in 7 of 10 active UC and 3 of 6 CD patients (NS). The cell count positive for IL-8 mRNA per unit area was definitely increased in moderate/severe UC when compared to mild UC (53.1 +/- 14.4/mm2 vs 9.0 +/- 5.1/mm2, P = 0.028) according to the degree of inflammation. IL-6 mRNA positive cells in CD were preferentially located in deeper lamina propria than IL-8 mRNA positive cells in UC. Interestingly, IL-8 mRNA was expressed in the mucosal epithelial cells in one UC patient. The patients treated by corticosteroids tended to show suppressed expression of each mRNA, except one patient with intractable UC. Our data suggest enhanced expression of mucosal IL-6 mRNA in CD and of IL-8 mRNA in UC by infiltrating mononuclear cells, indicating the distinct participation of each cytokine in the pathogenesis of UC and CD. Moreover, intestinal epithelial cells in UC occasionally exhibit IL-8 mRNA.

Topics: Colitis, Ulcerative; Crohn Disease; DNA Primers; DNA, Complementary; Gene Expression Regulation; Humans; Immunohistochemistry; In Situ Hybridization; Interleukin-6; Interleukin-8; Intestinal Mucosa; RNA, Messenger; Up-Regulation

Technetium Tc 99m hexamethyl propylene amine oxine (99mTc-HMPAO) has been used to radiolabel leukocytes with promising results for its clinical use in inflammatory bowel disease. During active ulcerative colitis, colonoscopy is indicated to determine the extent and the intensity of the disease for proper management. The aim of this prospective study was to determine whether 99mTc-HMPAO-labeled leukocyte scintigraphy can give information similar to that obtained with colonoscopy during acute attacks of ulcerative colitis.. Thirty-three consecutive patients with 50 acute episodes of ulcerative colitis underwent 99mTc-HMPAO scintigraphy and colonoscopy with biopsies. Scintigraphic determination of disease extent and intensity were compared with those obtained by colonoscopy with biopsies and clinicobiologic markers.. The scintigraphic index of disease intensity was correlated with endoscopic index, Truelove index, biologic markers, and local release of interleukin-8. The extent measured by scintigraphy was well correlated to the endoscopic and histologic extent.. 99mTc-HMPAO scintigraphy accurately determines the extent and the intensity of acute ulcerative colitis lesions. This noninvasive method can specify the extent and the intensity of an acute attack in patients with previously known ulcerative colitis.

Topics: Adult; Biomarkers; Colitis, Ulcerative; Colonoscopy; Female; Humans; Interleukin-8; Leukocytes; Male; Prospective Studies; Radionuclide Imaging; Radiopharmaceuticals; Sensitivity and Specificity; Severity of Illness Index; Technetium Tc 99m Exametazime

The pleiotropic cytokine leukemia inhibitory factor (LIF) possesses proinflammatory properties in common with tumor necrosis factor (TNF-alpha), interleukine (IL) -1 and -6, such as the induction of acute phase protein synthesis. LIF may have chemotactic activity through the induction of IL-8 production. LIF is produced by normal and tumoral cells and appears to facilitate in vivo rat colon carcinoma cells growth. Inflammatory bowel diseases, ulcerative colitis (UC) in particular, are histologically characterized by the infiltration of the colonic mucosa with activated neutrophils, macrophages and lymphocytes. Cytokines with their inflammatory as well as their regulatory activities may play a role in the perpetuation and possibly the initiation of inflammation in this disease and its local and/or systemic complications. Moreover, colorectal cancer is a late well identified complication in patients with long standing inflammatory bowel disease, UC in particular. Taken together, these results suggest that LIF could be involved in tumorigenic and/or metastatic processes of colorectal cells in UC patients. The aims of the present study was to quantify and to compare the colonic and systemic productions of LIF in UC patients. We showed for the first time in patients with UC, a high local production of LIF well correlated with IL-8 production. We also analyzed the effect of LIF on a human colon carcinoma cell line HT29. We demonstrated that LIF stimulated HT29 cell growth in a dose dependent-manner. These results suggest that LIF may play a critical role in the susceptibility of colonic host cells to tumor growth in patients with UC.

Topics: Adult; Animals; Caco-2 Cells; Case-Control Studies; Cell Division; Colitis, Ulcerative; Colonic Neoplasms; Female; Growth Inhibitors; HT29 Cells; Humans; Interleukin-6; Interleukin-8; Leukemia Inhibitory Factor; Lymphokines; Male; Rats; Recombinant Proteins

To study the effect of interleukin-1 beta and interleukin-8 in the pathogenesis of colitis and evaluate the therapeutic effect of interleukin-1 receptor antagonist(IL-1ra).. A rat model of chronic experimental colitis was induced by administration of trinitrobenzenesulfonic acid (TNBS, 30 mg) in 50% ethanol 0.25 ml. IL-1ra was then administered intravenously with a dosage of 7 mg/kg at different times. Hydrocortisone (3 mg) i.v. administration was served as control. Tissue expression of IL-1 beta mRNA and IL-8 mRNA was then studied by in situ hybridization before and after IL-1ra administration. Histological examination and tissue myeloperoxidase (MPO) and superoxide dismutase (SOD) activities detection were also made to assess the severity of colitis.. An acute inflammation with ulcers, neutrophil infiltration and crypt abscess developed that evolved into a chronic inflammation with abundant fibrous connective tissue hypertrophy at 21 days. And it then recurs only after 10 mg TNBS administration. Activities of MPO and SOD correlated with severity of inflammation. Expression of IL-1 beta mRNA was detected in macrophages in lamina propria and submucosa during the whole period of inflammation. It could be also present in epithelial cells in the 3rd day of colitis. While IL-8 mRNA expression appeared at the 3rd day, and disappeared at 21th day when colitis turned into chronic. After IL-1ra administration, the expression of these two interleukins could not be detected, accompanied with alleviation of histological manifestation, decrease of MPO activity and elevation of SOD activity.. IL-1 beta and IL-8 were involved in the pathogenesis of colitis. IL-1ra has preventive and therapeutic effect to colitis.

Topics: Animals; Colitis, Ulcerative; Ethanol; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-8; Male; Peroxidase; Rats; Rats, Wistar; RNA, Messenger; Sialoglycoproteins; Superoxide Dismutase; Trinitrobenzenesulfonic Acid

We examined the effect of ulinastatin in comparison with prednisolone (PSL) and salazosulfapyridine (SASP), well-known drugs for ulcerative colitis (UC), on two experimental UC models induced by dextran sulfate sodium (DSS) or trinitrobenzene sulfonic acid (TNB) in rats. Ulinastatin at the doses of 3000 approximately 10,000 units/kg/day (i.v.) significantly ameliorated the formation of erosion and infiltration of inflammatory cells in colonic mucosa in DSS-induced rat UC models. Moreover, ulinastatin at the dose of 10,000 units/kg/day (i.v.) significantly suppressed inflammation with ulcer in the colonic mucosa in TNB-induced rat UC models. PSL at the dose of 1 mg/kg/day (p.o.) also was as effective as ulinastatin on the above two UC models, while SASP at the dose of 100 mg/kg/day (p.o.) was less effective than ulinastatin and PSL. In addition, ulinastatin inhibited the activities of elastase and cathepsin G from human leukocytes, and it suppressed TNF alpha, IL-8 and superoxide production by rat macrophages and rabbit leukocytes in vitro. These results suggest that the suppression of inflammatory mediators produced by leukocytes is involved in the mechanism of the anti-UC action of ulinastatin.

Topics: Animals; Cathepsin G; Cathepsins; Colitis, Ulcerative; Female; Glycoproteins; Humans; Interleukin-8; Leukocyte Elastase; Leukocytes; Macrophages, Peritoneal; Male; Prednisolone; Rabbits; Rats; Rats, Sprague-Dawley; Rats, Wistar; Serine Endopeptidases; Sulfasalazine; Trypsin Inhibitors; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8) and nitric oxide (NO) may be important mediators in the pathogenesis of chronic idiopathic inflammatory bowel disease (CIIBD), but their roles in disease activity in ulcerative colitis (UC) and Crohn's disease (CD) are uncertain. The aim of this study was to measure mRNA for IL-8 and inducible NO synthase (iNOS) in small mucosal biopsies from untreated patients at first presentation and to relate these measurements to the histological levels of polymorph infiltration graded on a ten-point scale. For this purpose, a sensitive enzyme-linked oligonucleotide chemiluminescent assay (ELOCA) was developed to quantitate reverse transcription-polymerase chain reaction (RT-PCR) products amplified from RNA from paired biopsy samples. The levels of IL-8 and iNOS mRNAs were calculated as ratios of the RT-PCR products to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) RT-PCR product. In UC patients, median values of IL-8/GAPDH and iNOS/GAPDH were significantly elevated compared with controls and CD. However, in both UC and CD, the IL-8/GAPDH and iNOS/GAPDH ratios correlated significantly with polymorph infiltration. ELOCA enabled quantitation of multiple mRNAs in small mucosal biopsies from untreated patients with CIIBD and supported a role for IL-8 and iNOS in acute inflammation in both UC and CD.

Topics: Adult; Aged; Cell Movement; Colitis, Ulcerative; Crohn Disease; Female; Humans; Interleukin-8; Intestinal Mucosa; Luminescent Measurements; Male; Middle Aged; Neutrophils; Nitric Oxide Synthase; Polymerase Chain Reaction; RNA, Messenger

The aim of the study was to assess in vitro the production of interleukin 8 in Crohn's disease and ulcerative colitis.. Interleukin 8 concentrations (measured by ELISA) were evaluated in the culture supernatants of: a) peripheral blood mononuclear cells from 13 patients with Crohn's disease, 7 patients with ulcerative colitis and 7 controls, b) organ cultures of inflamed (22 Crohn's disease, 15 ulcerative colitis) and uninflamed (21 Crohn's disease, 9 ulcerative colitis, 15 controls) intestinal mucosal biopsies.. In patients with Crohn's disease and ulcerative colitis, the production of interleukin 8 by peripheral blood mononuclear cells and inflamed mucosa was higher than in controls (P < 0.01) without statistical difference between Crohn's disease and ulcerative colitis. Interleukin 8 production by normal mucosa was not different between the 3 groups. Interleukin 8 concentrations in the supernatants of organ culture were positively correlated to an endoscopical and a histological inflammatory index as well as to tumor necrosis factor, interleukin 1 and 6 concentrations.. These results support the notion that interleukin 8 plays an important role in the pathogenesis of inflammatory bowel disease.

Topics: Adult; Biopsy; Cells, Cultured; Colitis, Ulcerative; Crohn Disease; Female; Humans; Inflammation Mediators; Interleukin-8; Intestinal Mucosa; Leukocytes, Mononuclear; Male; Middle Aged; Organ Culture Techniques; Tumor Necrosis Factor-alpha

Inflammatory cytokines, including tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1 beta, have been implicated as primary mediators of intestinal inflammation in inflammatory bowel disease.. To investigate the in vitro effects of oxpentifylline (pentoxifylline; PTX; a phosphodiesterase inhibitor) on inflammatory cytokine production (1) by peripheral mononuclear cells (PBMCs) and (2) by inflamed intestinal mucosa cultures from patients with Crohn's disease and patients with ulcerative colitis.. PBMCs and mucosal biopsy specimens were cultured for 24 hours in the absence or presence of PTX (up to 100 micrograms/ml), and the secretion of TNF-alpha, IL-1 beta, IL-6, and IL-8 determined by enzyme linked immunosorbent assays (ELISAs).. PTX inhibited the release of TNF-alpha by PBMCs from patients with inflammatory bowel disease and the secretion of TNF-alpha and IL-1 beta by organ cultures of inflamed mucosa from the same patients. Secretion of TNF-alpha by PBMCs was inhibited by about 50% at a PTX concentration of 25 micrograms/ml (IC50). PTX was equally potent in cultures from controls, patients with Crohn's disease, and those with ulcerative colitis. The concentrations of IL-6 and IL-8 were not significantly modified in PBMCs, but IL-6 increased slightly in organ culture supernatants.. PTX or more potent related compounds may represent a new family of cytokine inhibitors, potentially interesting for treatment of inflammatory bowel disease.

Topics: Adult; Colitis, Ulcerative; Colon; Crohn Disease; Culture Techniques; Cytokines; Female; Humans; Inflammatory Bowel Diseases; Interleukin-1; Interleukin-6; Interleukin-8; Intestinal Mucosa; Leukocytes, Mononuclear; Male; Middle Aged; Pentoxifylline; Phosphodiesterase Inhibitors; Tumor Necrosis Factor-alpha

Epithelial cell-derived neutrophil-activating protein-78 (ENA-78) is a neutrophil-directed C-X-C chemokine. We report that Caco-2 and T84 human intestinal epithelial cells produce ENA-78 after stimulation by interleukin (IL)-1 beta or tumor necrosis factor-alpha. Caco-2 cells show increased IL-8 production at 4-12 h and increased ENA-78 production at 8-24 h after cytokine stimulation. Immunohistochemical studies in normal human colon and in ulcerative colitis demonstrate ENA-78 immunoreactivity principally associated with crypt epithelial cells. Furthermore, human colonic tissues from patients with ulcerative colitis show elevated levels of ENA-78 mRNA (24-fold increase, P < 0.01) and protein (4-fold increase, P < 0.05) compared with normal controls. Thus ENA-78 is produced in normal colon and in ulcerative colitis and is predominantly of enterocyte origin. The kinetics of ENA-78 induction in human colon epithelial cell lines are delayed and prolonged compared with IL-8. We propose that ENA-78 and IL-8 serve complementary and sequential roles in neutrophil recruitment in ulcerative colitis. ENA-78 as an enterocyte-derived, neutrophil-activating chemokine may be especially important in neutrophil recruitment from the lamina propria into the epithelial layer.

Topics: Cell Line; Chemokine CXCL5; Chemokines, CXC; Colitis, Ulcerative; Colon; Colonic Neoplasms; Crohn Disease; DNA Primers; Enzyme-Linked Immunosorbent Assay; Humans; Immunohistochemistry; Interleukin-1; Interleukin-8; Intestinal Mucosa; Polymerase Chain Reaction; Protein Biosynthesis; RNA, Messenger; Signal Transduction; Transcription, Genetic; Tumor Cells, Cultured

Interleukin-8 (IL-8) is an important cytokine for recruitment and activation of polymorphonuclear neutrophils (PMNs), cells that are abundant in the intestinal lesions of ulcerative colitis (UC) and Crohn's disease (CD). The present investigation was conducted to evaluate intestinal IL-8 concentration and IL-8 gene expression in parallel in inflammatory bowel disease (IBD) patients and a non-inflammatory control group.. The intestinal concentration of IL-8 was measured with a sandwich enzyme-linked immunosorbent assay (ELISA) technique (detection limit, 17.4 pg/mg protein), and relative quantitation of IL-8 mRNA transcript levels was done with a reverse transcription polymerase chain reaction (RT-PCR)-based method. Biopsy specimens from 66 humans who underwent colonoscopy--28 with UC, 18 with CD and colonic involvement, and 20 non-inflammatory disease-specific controls who subsequently were found to fulfill the diagnostic criteria for irritable bowel syndrome (IBS)--were included. None had received glucocorticoids within 3 months.. Using a one-tailed variance analysis, a significant concordance between increasing IL-8 protein concentrations and disease activity was found both in UC and CD (P < 0.001), and only trace amounts were detected in IBS biopsy specimens. No differences were found between the two groups of UC and CD patients (P > 0.05), and no differences were found between quiescent IBD and IBS (P > 0.05). However, the PCR method showed IL-8 mRNA in 8 of 18 CD patients (44.4%; 95% confidence limits, 21.5-69.2%) and 7 of 28 UC patients (25.9%; 95% confidence limits, 11.1-46.3%), as compared with 0 of 20 IBS (P < 0.005). Increased IL-8 mRNA levels were found only in active CD, which was not the case in UC. No correlation was found between intestinal IL-8 ELISA and IL-8-mRNA levels (r = 0.24, P > 0.05).. The observed correlation between disease activity and expression of the IL-8 gene in active CD colitis but not in UC and the increased IL-8 protein concentrations in affected intestinal segments of IBD as compared with the non-inflamed IBS indicate a possible transient IL-8 gene expression or altered mRNA stability in UC and CD, as is well known for other cytokines, such as IL-2. If so, it may form the basis of new therapeutic regimens for IBD like IL-10.

Topics: Adult; Case-Control Studies; Colitis, Ulcerative; Colon; Crohn Disease; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Polymerase Chain Reaction; RNA, Messenger

Cell surface adhesion molecules (CAM) are important promotors of the immunoinflammatory cascade. The circulating levels of soluble intercellular adhesion molecule 1 (ICAM-1) have previously been shown to correlate with disease activity in inflammatory bowel disease. The primary aim of this study was consequently to investigate if this also applies to mucosal levels of soluble ICAM-1. We measured soluble ICAM-1 levels in intestinal biopsy specimens and the endoscopic activity of 69 patients with ulcerative colitis (UC) and 14 controls and found that the median concentration of soluble ICAM-1 was significantly higher in patients with moderately or very active UC (15.0 ng/ml) as compared to slightly active (9.8 ng/ml) and inactive UC (9.5 ng/ml) as well as controls (6.5 ng/ml) (P < 0.005). To further elucidate the interactions, two other CAM [E-selectin and vascular cellular adhesion molecule 1 (VCAM-1)], together with interleukin-8 (IL-8), IL-2 receptor (IL-2R) alpha and beta chains, were also measured. A significant trend towards higher soluble E-selectin levels in biopsies with active UC (1.8 pg/ml) as compared to inactive UC (1.3 pg/ml) and to controls (< 1.0 pg/ml) (P < 0.01) was also found. In contrast, soluble VCAM-1 was barely detectable in biopsies from two UC patients. A significant correlation was found between soluble ICAM-1 and IL-8 concentrations (r = 0.46; P < 0.0001), and between sICAM-1 and sIL-2R alpha concentrations (r = 0.69; P < 0.0001), while sIL-2R beta was not detected. This study shows that intestinal ICAM-1 and E-selectin correlate with endoscopic activity of UC and with IL-8 and IL-2R alpha levels. These mediators may be useful in monitoring mucosal inflammation in studies exploring the therapeutical potential of targeting CAM. The lack of detectable VCAM-1, which is induced only in venous endothelium is interesting. It may suggest that intestinal inflammation mainly affects arterial endothelial cells and support the theory that intestinal vasculitis is involved in the pathogenesis of inflammatory bowel disease.

Topics: Acute Disease; Adult; Aged; Cell Adhesion Molecules; Colitis, Ulcerative; Colonic Diseases, Functional; E-Selectin; Female; Humans; Interleukin-8; Male; Middle Aged; Vascular Cell Adhesion Molecule-1

To test whether there is a difference in the expression of interleukin 8 (IL8) between Crohn's disease and ulcerative colitis and to determine the main site of its synthesis this study analysed IL8 in mucosal biopsy specimens of patients with Crohn's disease and ulcerative colitis by enzyme linked immunosorbent assay (ELISA) and by in situ hybridisation. IL8 was measured by ELISA in 38 normal control patients, eight inflammatory control patients, 55 Crohn's disease biopsy specimens (26 patients), and 67 ulcerative colitis biopsy specimens (35 patients). IL8 mRNA was determined in samples by in situ hybridisation using a specific IL8 RNA probe. IL8 protein was significantly increased in macroscopically inflamed specimens of Crohn's disease (median 118 pg/specimen, p < 0.0001), ulcerative colitis (median 140 pg/specimen, p < 0.001), and inflammatory controls (median 30 pg/specimen, p = 0.010) compared with normal controls (median 4 pg/specimen). IL8 was also increased in uninflamed specimens of Crohn's disease (median 46 pg/specimen, p < 0.001) but not of ulcerative colitis patients (median 9 pg/specimen, p = 0.3). IL8 protein in the mucosa correlated significantly with macroscopic inflammation in Crohn's disease (r = 0.47, p < 0.001) and in ulcerative colitis (r = 0.60, p < 0.001). IL8 mRNA was detected by in situ hybridisation in 31 of 55 biopsy specimens (56%) of Crohn's disease patients, in 38 of 67 specimens of ulcerative colitis patients (57%), in five of eight inflammatory controls (63%) and in five of 38 normal controls (13%). Mucosal IL8 mRNA expression correlated with mucosal IL8 protein (r = 0.46, p < 0.001). IL8 mRNA was only detected in inflammatory cells of the interstitium but not in mucosal epithelial cells. IL8 is produced mainly in the lamina propria of the colon in inflammatory bowel disease and correlates with mucosal inflammation.

Topics: Adolescent; Adult; Aged; Biopsy; Case-Control Studies; Colitis, Ulcerative; Colon; Crohn Disease; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression; Humans; In Situ Hybridization; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; RNA, Messenger

The movement of neutrophils into the colonic mucosa in ulcerative colitis is induced by chemokines including interleukin 8 (IL8) and leukotriene B4 (LTB4).. To compare the ability of mucosa from ulcerative colitis patients and controls to stimulate neutrophil movement, to define the contribution of LTB4 to this, and to define the relative biological importance of LTB4 and IL8.. Resected mucosa was obtained from seven control patients and 10 patients with ulcerative colitis.. Mucosal homogenate supernatants were used to stimulate isolated neutrophils and the effect assessed by the neutrophil shape change response. Responses were inhibited with either the LTB4 receptor antagonist SC41930- or neutralising anti-IL8 antibody. LTB4 was extracted and assayed by RIA.. Homogenate supernatants from inflamed mucosa were more bioactive (median 1.2 mg/ml-1 induced 50% response) than those from uninflamed mucosa (4.25 mg/ml-1 induced 50% response; difference 2.8 mg/ml-1 (96.5% CI 0.5 to 6.1, p < 0.05). Maximal inhibition by SC41930 of the response was significantly greater in inflamed mucosa (54% median) than in uninflamed mucosa (34%). This inhibition correlated with the level of immunoreactive LTB4 (r = 0.78). Anti-IL8 reduced bioactivity of homogenate supernatants from inflamed mucosa (at 1:10 dilution) by 11.4% (IQR 1.2 to 51.8, p = 0.021) whereas SC41930 reduced it by 54.8% (35.6 to 77.5, p = 0.008).. Inflamed colonic mucosa releases more neutrophil movement inducing bioactivity than uninflamed mucosa, and has greater LTB4 dependent activity. It yields both IL8 and LTB4 dependent activity but greater LTB4 dependent activity.

Topics: Adult; Aged; Aged, 80 and over; Benzopyrans; Case-Control Studies; Cell Movement; Colitis, Ulcerative; Female; Globulins; Humans; Interleukin-8; Intestinal Mucosa; Leukotriene B4; Male; Middle Aged; Neutrophils; Radioimmunoassay

Epithelia from several sites exhibit inducible secretion of interleukin 8 (IL-8). This study aimed to characterise secretion of IL-8 by colonic epithelial cells in vitro. Colonic crypt cells were isolated enzymatically from resected colon and the IL-8 content of culture supernates was measured by ELISA. The rate of secretion of IL-8 accelerated and levels of IL-8 transcripts increased appreciably during culture. Exposure to tumour necrosis factor alpha (TNF alpha) failed to increase secretion further. Secretion was not induced by the enzymatic digestion or by serum used in the culture medium but was significantly inhibited by butyrate, by a mean of 23%. Control experiments indicated that colonic crypt cells were the likely source. The secretion of IL-8 over 24 hours by cells from uninflamed mucosa of patients with ulcerative colitis or Crohn's disease was more than twofold that from normal cells, while that from cancer bearing colons was normal. TNF alpha (10 mM) significantly suppressed IL-8 secretion only in the ulcerative colitis group and the change was different to those in the normal (p = 0.007) and Crohn's disease groups (p = 0.012). Cells from inflamed areas secreted more IL-8 than those from autologous uninflamed areas (p = 0.009) but responses to modulating factors were no different. The induction of IL-8 secretion by colonic crypt cells in vitro is probably a response to injury associated with isolation and culture. It is suppressed by butyrate and increased in inflammatory bowel disease independently of the presence of mucosal inflammation. Whether epithelial derived IL-8 plays a part in the pathogenesis of inflammatory bowel disease is not yet clear.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Blotting, Northern; Butyrates; Cell Survival; Cells, Cultured; Colitis, Ulcerative; Colon; Crohn Disease; Female; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Tumor Necrosis Factor-alpha

Topics: Antibodies, Monoclonal; Colitis, Ulcerative; Colon; Gene Expression; Humans; Immunity, Mucosal; Immunohistochemistry; In Situ Hybridization; Interleukin-1; Interleukin-8; Intestinal Mucosa; Lymphocyte Activation; RNA, Messenger; T-Lymphocytes

Controversy exists as to whether pouchitis represents a reactivation of the immunologic mechanisms that lead to ulcerative colitis (UC). The aims of this study were to determine local levels of the cytokines: interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor alpha (TNF alpha) in the mucosa of patients with "asymptomatic" ileoanal pouch (n = 25), pouchitis (n = 9), active UC (n = 20), normal ileum (n = 15), proctitis (n = 10), and normal colon (n = 15).. Lamina propria mononuclear cells were isolated from mucosal biopsies by enzymatic dispersion and cultured for 48 hours. Proinflammatory cytokine levels were measured in the supernatants by enzyme-linked immunosorbent assay.. IL-1 beta, IL-6, IL-8, and TNF alpha secretions were significantly greater in pouchitis and active UC than in the noninflamed ileoanal pouch and normal controls (P < 0.001). There was significant correlation (r = 0.63, P < 0.05) between levels of cytokines expressed in pouchitis and active UC.. Increased cytokine expression occurs in both active UC and pouchitis and to a lesser extent in the long-standing ileoanal pouch.

Topics: Adolescent; Adult; Aged; C-Reactive Protein; Case-Control Studies; Colitis, Ulcerative; Colon; Humans; Ileitis; Ileum; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Intestinal Mucosa; Leukocytes, Mononuclear; Middle Aged; Orosomucoid; Proctitis; Proctocolectomy, Restorative; Tumor Necrosis Factor-alpha

The etiology of ulcerative colitis (UC) remains uncertain. It has been said that in patients with an active phase of UC the levels of the proinflammatory cytokines increased. Paying particular attention to patients with inactive UC, we investigated several cytokines' m-RNA transcripts.. In 27 patients with UC, the mucosal cytokine expression of m-RNA including IL-8, TNF-alpha, IFN-gamma and IL-6 were examined using the reverse transcription-coupled polymerase chain reaction (RT-PCR) and oligonucleotide hybridization.. 1) The expression ratios of m-RNA of IL-8, IFN-gamma and TNF-alpha in cases with UC (92.6%, 70.4% and 77.8%, respectively) were significantly higher than in normal controls (17.6%, 0.0% and 5.9%, respectively) (p < 0.01). Regarding IL-8 and IFN-gamma, those in UC cases were also significantly higher than in cases with other types of colitis (p < 0.05). 2) No significant difference was noted between active and inactive cases with UC in terms of the expression ratios of m-RNA of IL-8, TNF-alpha and IFN-gamma. 3) Only the expression ratio of IL-6 m-RNA correlated with the macroscopic score according to Matts. In addition, expression of IL-6 m-RNA in the colon tissue was observed in 9 out of 10 cases with other types of colitis (90.0%).. The expression of IL-8, TNF-alpha and IFN-gamma m-RNA increased even in inactive UC, suggesting that these three cytokines play an important role in the pathogenesis of repeated inflammation of UC.

Topics: Adult; Aged; Base Sequence; Colitis, Ulcerative; Electrophoresis, Agar Gel; Female; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-6; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Molecular Sequence Data; Nucleic Acid Hybridization; RNA, Messenger; Tumor Necrosis Factor-alpha

Tissue and plasma concentrations of several cytokines are increased in patients with inflammatory bowel disease (IBD). Platelets play an important role in inflammation and circulate in an activated state in patients with IBD. This study assesses the expression of IL-8 and IL-1 receptors on the surface of platelets from patients with IBD using phycoerythrin (PE)-labelled recombinant human rhIL-1 beta and rhIL-8 and flow cytometry. The percentage IL-1R expressing platelets (median and interquartile range IQR) in the IBD group was 8.7% (5.5-18.2) compared to 4.2% (2.3-6.1) in controls (P = 0.02). The percentage IL-8R expressing platelets in the IBD group was 22.5% (16.5-27.9) and 9.2% (4.3-9.6) in controls (P < 0.001). Furthermore, platelet IL-1R expression in patients with IBD was inversely related to the total daily dose of steroids (r = 0.71, P < 0.01 linear regression analysis). Finally, platelet rich plasma from healthy controls was stimulated with rhIL-1 beta and rhIL-8 and assessed for activation dependent expression of platelet aGPIIb/IIIa and CD62 (p-selectin, GMP-140). IL-1 beta and IL-8 in vitro significantly and specifically activated the platelets. The surface membrane of platelets is able to express functional IL-1R and IL-8R, the expression of which is significantly increased in IBD. Interleukin-1 beta and IL-8 modulate platelet activation in vitro indicating a target role for platelet function in inflammation.

Topics: Adolescent; Adult; Blood Platelets; Colitis, Ulcerative; Crohn Disease; Female; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; P-Selectin; Platelet Activation; Platelet Membrane Glycoproteins; Receptors, Interleukin; Receptors, Interleukin-1; Receptors, Interleukin-8A

IL-8 is generating increasing interest as a powerful neutrophil chemoattractant and activator. To elucidate the mechanisms of neutrophil infiltration in inflammatory bowel disease, we examined 33 patients with ulcerative colitis (UC), 18 with Crohn's disease (CD), eight with some other type of colitis, and 18 normal control subjects for measurement of IL-8 in homogenates of colonic biopsy specimens. The affected colonic mucosa was found to contain significantly more IL-8 in patients with active inflammatory bowel disease than in patients with inactive disease (UC, P < 0.001; CD, P < 0.001), in patients with other types of colitis (UC, P < 0.05; CD, P < 0.01), or in normal control subjects (UC, P < 0.001; CD, P < 0.001). Colonic IL-8 levels correlated significantly with the macroscopic grade of local inflammation, especially in patients with UC (P < 0.001). Colonic IL-8 levels also correlated well with the neutrophil numbers in mucosal tissue (UC, r = 0.950, P < 0.001; CD, r = 0.940, P < 0.001), and with colonic IL-1 beta (r = 0.911, P < 0.001) and tumour necrosis factor-alpha (TNF-alpha) levels (r = 0.604, P < 0.001) in patients with these two conditions. These data suggest a potential role for IL-8 and its regulatory cytokines IL-1 and TNF-alpha in mediating neutrophil infiltration of the gut wall in inflammatory bowel disease.

Topics: Adolescent; Adult; Aged; Cell Count; Chemotactic Factors; Colitis, Ulcerative; Crohn Disease; Female; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Neutrophils; Tumor Necrosis Factor-alpha

Concentrations of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-alpha) were determined by solid-phase ELISA in tissue homogenates of mucosal biopsy specimens obtained from pelvic ileal pouches in 13 patients with pouchitis (reservoir ileitis) and 17 with pouches without pouchitis. Normal ileal mucosa was used as a control. IL-1 beta was detected in all tissue homogenates from patients with pouchitis compared with only 29% from pouches without pouchitis and none from controls. IL-6 and IL-8 were present in all pouchitis specimens, in 70% of the specimens from nonpouchitis and only 30% of specimens from controls. TNF-alpha was undetectable in all specimens examined. The concentrations of IL-1 beta, IL-6, and IL-8 were significantly greater (P < 0.001) in biopsy specimens from pouchitis compared to those from pouches without pouchitis or normal ileal mucosa and in patients with pouchitis tissue levels of IL-1 beta significantly correlated with IL-6 (P < 0.05) and IL-8 (P < 0.01). Furthermore IL-1 and IL-8 levels were significantly higher in tissue specimens from nonpouchitis pouches than in those from normal ileal mucosa (P < 0.02). These results suggest that an enhanced cellular immunity operates in vivo at the mucosal level in pouchitis as in the case of ulcerative colitis.

Topics: Adult; Colitis, Ulcerative; Enzyme-Linked Immunosorbent Assay; Female; Humans; Ileum; Inflammation; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Intestinal Mucosa; Male; Proctocolectomy, Restorative; Tumor Necrosis Factor-alpha

A quantitative polymerase chain reaction (PCR) assay for mRNA expression of interleukin-8 (IL-8), a neutrophil chemotactant and activator, was developed to examine the expression of this cytokine by colonic mucosa. A synthetic IL-8 RNA deleted in size of native IL-8 mRNA was used as an external control. The synthetic IL-8 RNA was mixed with total RNA from cells and converted to cDNA and amplified by PCR simultaneously. The lower limit of sensitivity for the assay was found to be more than 1 femtogram of IL-8 mRNA. The assay determined IL-8 mRNA expression when the RNA was isolated from either human histiocytic lymphoma cell line U937 cells or human colonic mucosa obtained from colitis patients and healthy controls. The development of the rapid and sensitive assay should provide a means to more fully evaluate the role of this cytokine in diverse disease states with small scale.

Topics: Base Sequence; Colitis, Ulcerative; DNA Primers; Electrophoresis, Agar Gel; Humans; Interleukin-8; Intestinal Mucosa; Lymphoma, Large B-Cell, Diffuse; Molecular Sequence Data; Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured

Interleukin-8 (IL-8) is a potent cytokine for recruitment and activation of neutrophils. To visualize its distribution in the intestinal mucosa and to understand better its possible role in the induction and promotion of inflammatory bowel disease, expression of the IL-8 gene was analyzed in resected bowel segments of 14 patients with active Crohn's disease or ulcerative colitis. In situ hybridization with IL-8 anti-sense RNA probes revealed strong and specific signals in the histologically affected mucosa. The number of cells expressing IL-8 gene correlated with the histological grade of active inflammation. In accordance with the characteristic histological signs of active disease, IL-8-expressing cells were diffusely distributed over the entire affected mucosa in patients with ulcerative colitis, whereas in patients with Crohn's disease, IL-8-expressing cells showed a focal distribution pattern. Cells expressing IL-8 were mainly located at the base of ulcers, in inflammatory exudates on mucosal surfaces, in crypt abscesses, and at the border of fistulae. Analysis of semi-serial sections pointed to macrophages, neutrophils, and epithelial cells as possible sources of this cytokine in active inflammatory bowel disease. We consistently failed to detect IL-8 messenger RNA in the mucosa of uninvolved bowel segments and in normal-appearing control mucosa of patients with colon cancer. In contrast, tissue specimens from two patients with acute appendicitis displayed IL-8-expressing cells in the mucosa. These results support the notion that IL-8 plays and important but nonspecific role in the pathogenesis of inflammatory bowel disease and that the production of IL-8 messenger RNA is restricted to areas with histological signs of inflammatory activity and mucosal destruction.

Topics: Adult; Aged; Colitis, Ulcerative; Crohn Disease; Female; Gene Expression Regulation; Humans; In Situ Hybridization; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; RNA Probes; Severity of Illness Index

The concentration of myeloperoxidase, a neutrophil granule constituent, was measured in the perfusion fluid from sigmoid and rectal segments in patients with ulcerative colitis. The concentrations of myeloperoxidase were increased severalfold in the patients with ulcerative colitis compared with healthy controls pointing to an enhanced neutrophil activity. The release of myeloperoxidase correlated to an enhanced local release of the neutrophil activating peptide interleukin-8 (IL-8). Increased values of tumour necrosis factor (TNF-alpha) were also found during intestinal perfusion of the patients and correlated with those of IL-8. The results obtained are compatible with the hypothesis that local mucosal recruitment/activation of neutrophils in ulcerative colitis is mediated by an enhanced IL-8 synthesis. TNF-alpha may be one relevant factor as a stimulus to IL-8 synthesis.

Topics: Adult; Aged; Colitis, Ulcerative; Colon; Female; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Neutrophils; Perfusion; Peroxidase; Tumor Necrosis Factor-alpha

1. We studied neutrophil-activating peptide-1/interleukin-8 in inflammatory bowel disease. 2. Mucosal levels of neutrophil-activating peptide-1/interleukin-8 were significantly higher in patients with active ulcerative colitis [median 74.5 (range 17.7-450.8) pg/mg] than in patients with active Crohn's disease [10.4 (4-46.9) pg/mg; P less than 0.002] or in normal control subjects [10.4 (4-16.6) pg/mg; P less than 0.002]. 3. Circulating neutrophil-activating peptide-1/interleukin-8 was generally undetectable but there were higher levels of anti-neutrophil-activating peptide-1/interleukin-8 antibodies in patients with active ulcerative colitis [62.9 (3.4-239) ng/ml] than in patients with active Crohn's disease [5.9 (2.1-18.10) ng/ml; P less than 0.001] or in control subjects [6.1 (3.2-15.8) ng/ml; P less than 0.001]. 4. Neutrophil-activating peptide-1/interleukin-8 may be of specific functional importance in mediating inflammation in ulcerative colitis.

Topics: Antibodies; Colitis, Ulcerative; Crohn Disease; Humans; Interleukin-8; Intestinal Mucosa

In this study, mediators of inflammation were characterized in colonic and terminal ileum mucosa from subjects with ulcerative colitis. We considered the role of two different chemotactic factors (interleukin-8 and leukotriene B4) and of myeloperoxidase in the pathology of inflammatory bowel disease. Serial biopsy specimens were taken at different sites, washed in 0.02 M phosphate/saline buffer, homogenized, and then sonically disrupted. In both the proximal and distal regions of the colonic mucosa of ulcerative colitis patients, there was a more than 10-fold increase in interleukin-8 levels over that in control subjects (> 300 pg/mg protein vs. 30 pg/mg protein in controls, p < or = 0.01). However, terminal ileum levels of interleukin-8 were the same in ulcerative colitis and control groups (150 pg/mg protein). There was also a 3- to 5-fold increase in leukotriene B4 levels and a several-fold increase in myeloperoxidase levels throughout the colonic mucosa in patients with ulcerative colitis. This study demonstrates that 1) interleukin-8 may have an immunoregulatory role in the pathogenesis of inflammatory bowel disease, and 2) interleukin-8, myeloperoxidase, and leukotriene B4 may be useful markers for the biochemical identification of inflammatory bowel disease.

Topics: Adult; Aged; Biomarkers; Catalase; Colitis, Ulcerative; Colon; Humans; Ileum; Interleukin-8; Intestinal Mucosa; Leukotriene B4; Middle Aged; Neutrophils; Peroxidase; Radioimmunoassay