Different Stages Of Ulcerative Colitis Research Articles

Using serum peptidomics to discovery the diagnostic marker for different stage of ulcerative colitis

The use of peptidomics to find diagnostic markers has attracted increasing clinical attention. Ulcerative colitis (UC) is a type of inflammatory bowel disease, and the traditional auxiliary diagnostic technique is colonoscopy. However, this invasive method is not effective in distinguishing between patients with endoscopic remission and healthy people, which carries the risk of delayed diagnosis of UC. In this study, we used peptidomics to find serum diagnostic markers for different stages of UC. A total of 78 serum samples were collected to form a training set (60 samples) and a testing set (18 samples). Among them, patients with active UC, remitting UC and healthy people accounted for one third each. The nano-liquid chromatography coupled with hybrid linear trap quadrupole orbitrap mass spectrometry was used for detection of low molecular weight peptides in serum. According to the protein database search and de novo sequencing algorithm, forty peptides were simultaneously identified in all samples. Six biomarker peptides were screened in the training set through orthogonal partial least-squares-discriminant analysis and receiver operating characteristic curve analysis. These six peptides were derived from proteins involved in coagulation and complement activation. We evaluated the diagnostic ability of the six peptides in the testing set through hierarchical cluster analysis, and showed that perturbation of these peptides could distinguish patients with active UC, patients with remitting UC and healthy people. This study validated the feasibility of serum peptidomics for the discovery of diagnostic markers, and provided a potential method for diagnosing different stages of UC.

Inhibition of C3a/C3aR Axis in Diverse Stages of Ulcerative Colitis Affected the Prognosis of UC by Modulating the Pyroptosis and Expression of Caspase-11.

Ulcerative colitis (UC) is a serious digestive system disease. Furthermore, the activation of C3a/C3aR axis promoted the expression of caspase-11. And higher levels of caspase-11 could induce the pyroptosis and inflammation of cells. However, some studies suggested that caspase-11 could promote and suppress the inflammation during the development of UC. In addition, whether C3a/C3aR axis could affect the development of UC by modulating the expression of caspase-11 is unclear. We established the UC rat model in this study. Next, the C3aR inhibitor was used to treat these rats at diverse stages of UC. Next, the HE staining was performed to detect the intestinal damage. ELISA was performed to reveal the expression of IL-6 and TNF-α in different stages of UC. Western blotting was used to detect the expression of caspase-11 and C3aR in different stages of UC. Stimulation of C3aR inhibitor in early stage of UC promoted the expression of C3aR and caspase-11 in later stage of UC. Treatment of C3aR inhibitor in later stage of UC inhibited the expression of C3aR and caspase-11 in later stage of UC. Furthermore, application of C3aR inhibitor in early stage of UC aggravates the damage of colon tissue and enhanced the secretion of TNF-α and IL-6 in the later stage of UC. Treatment of C3aR inhibitor in later stage of UC relieved the symptoms of UC and suppressed the production of TNF-α and IL-6 in the later stage of UC. Application of C3aR inhibitor in early stage of UC induced the poor prognosis of UC by upregulating the expression of caspase-11. Treatment of C3aR inhibitor in later stage of UC relieved the symptoms of UC and lead to the favorable prognosis of UC by inhibiting the expression of caspase-11.

Relationship between the expression of NOD-like receptor protein 3 inflammasome and the activity of ulcerative colitis

Objective To investigate the relationship between the expression of NOD-like receptor protein 3 (NLRP3) inflammasome in colonic mucosal tissues of ulcerative colitis (UC) patients and UC mice model and the activity of UC. Methods From December 2016 to January 2018, at Department of Gastroenterology, the First Affiliated Hospital of Zhejiang Traditional Chinese Medicine University, 60 patients with UC were recruited, of which 15 cases at remission phase, 15 cases at mild activity phase, and 15 cases at moderate activity phase, and 15 cases at severe activity phase; and 15 healthy subjects were selected as healthy control group. UC mice models were established by dextran sulfate sodium (DSS). Forty-eight BALB/c mice were divided into 2.5% DSS group, 5.0% DSS group and 7.5% DSS group and control group. The colon tissues of UC patients and UC mice models were pathologically scored. The expression of NLRP3, cysteinyl aspartate specific proteinase 1(caspase-1) and apoptosis-associated speck-like protein containing CARD (ASC) at mRNA level in colon tissues of UC patients and UC mice models were determined by real time fluorescence quantitative polymerase chain reaction (PCR). The expression of NLRP3, caspase-1 and ASC at protein level in colon tissues of UC patients and UC mice models were detected by Western blotting. One-way analysis of variance and SNK-t test were performed for statistical analysis. Results The histopathological scores of colon tissues of UC patients at remission phase, at mild activity phase, at moderate activity phase, at severe activity phase and healthy controls were 2.37±0.46, 4.84±1.29, 6.82±0.96, 9.42±1.13 and 1.23±0.55, respectively; the differences were statistically significant (F=67.68, P<0.01). The higher the degree of inflammation, the higher the pathological score, and the differences were statistically significant (all P<0.05). The pathological scores of colon tissues of mice in the 2.5% DSS group, 5.0% DSS group and 7.5% DSS group were 4.54±0.74, 6.02±1.00 and 8.43±1.46, respectively; the higher the dose of DSS, the higher the pathological score, and the differences were statistically significant (all P<0.05). The expression of NLRP3 at mRNA level of UC at remission phase, mild activity phase, moderate activity phase, severe activity phase and healthy controls were 1.15±0.10, 1.49±0.13, 2.00±0.25, 2.05±0.33 and 0.61±0.09, respectively; the expression of caspase-1 at mRNA level were 1.13±0.08, 1.51±0.19, 2.10±0.23, 2.88±0.33 and 0.61±0.11, respectively; the expression of ASC at mRNA level were 1.12±0.08, 1.88±0.33, 2.53±0.22, 3.20±0.24 and 0.59±0.12, respectively; the differences between groups were statistically significant (F=108.43, 63.25 and 105.25, all P<0.01). The higher the degree of inflammation, the higher the mRNA expression levels of NLRP3, caspase-1 and ASC, and the differences were statistically significant (all P<0.01). The higher the dose of DSS, the higher the protein expression levels of NLRP3, caspase-1 and ASC at mRNA level. The higher the degree of inflammation, the higher expression of NLRP3, caspase-1 and ASC in colon tissues of UC patients. The higher dose of DSS, the higher the protein expression levels of NLRP3, caspase-1 and ASC in colon tissues of mice. Conclusions The expression level of NLRP3 inflammasome is different in different stages of UC, the higher degree of inflammatory activity, the higher the expressie level. It is helpful to evaluate the activity of UC by detecting the expression level of NLRP3 inflammasome. Key words: Ulcerative colitis; NLRP3 inflammasome; Activity of inflammation

Molecular profiling of mucosal tissue associated microbiota in patients manifesting acute exacerbations and remission stage of ulcerative colitis.

Dysbiosis of intestinal microflora has been postulated in ulcerative colitis (UC), which is characterized by imbalance of mucosal tissue associated bacterial communities. However, the specific changes in mucosal microflora during different stages of UC are still unknown. The aim of the current study was to investigate the changes in mucosal tissue associated microbiota during acute exacerbations and remission stages of UC. The mucosal microbiota associated with colon biopsy of 12 patients suffering from UC (exacerbated stage) and the follow-up samples from the same patients (remission stage) as well as non-IBD subjects was studied using 16S rRNA gene-based sequencing and quantitative PCR. The total bacterial count in patients suffering from exacerbated phase of UC was observed to be two fold lower compared to that of the non-IBD subjects (p = 0.0049, Wilcox on matched-pairs signed rank tests). Bacterial genera including Stenotrophomonas, Parabacteroides, Elizabethkingia, Pseudomonas, Micrococcus, Ochrobactrum and Achromobacter were significantly higher in abundance during exacerbated phase of UC as compared to remission phase. The alterations in bacterial diversity with an increase in the abnormal microbial communities signify the extent of dysbiosis in mucosal microbiota in patients suffering from UC. Our study helps in identifying the specific genera dominating the microbiota during the disease and thus lays a basis for further investigation of the possible role of these bacteria in pathogenesis of UC.

Altered Colonic Mucosal Polyunsaturated Fatty Acid (PUFA) Derived Lipid Mediators in Ulcerative Colitis: New Insight into Relationship with Disease Activity and Pathophysiology

ObjectivesUlcerative colitis (UC) is a relapsing inflammatory disorder of unconfirmed aetiology, variable severity and clinical course, characterised by progressive histological inflammation and with elevation of eicosanoids which have a known pathophysiological role in inflammation. Therapeutic interventions targetting eicosanoids (5-aminosalicylates (ASA)) are effective first line and adjunctive treatments in mild-moderate UC for achieving and sustaining clinical remission. However, the variable clinical response to 5-ASA and frequent deterioration in response to cyclo-oxygenase (COX) inhibitors, has prompted an in depth simultaneous evaluation of multiple lipid mediators (including eicosanoids) within the inflammatory milieu in UC. We hypothesised that severity of inflammation is associated with alteration of lipid mediators, in relapsing UC.DesignStudy was case-control design. Mucosal lipid mediators were determined by LC-MS/MS lipidomics analysis on mucosal biopsies taken from patients attending outpatients with relapsing UC. Univariate and multivariate statistical analyses were used to investigate the association of mucosal lipid mediators, with the disease state and severity graded histologically.ResultsLevels of PGE2, PGD2, TXB2, 5-HETE, 11-HETE, 12-HETE and 15-HETE are significantly elevated in inflamed mucosa and correlate with severity of inflammation, determined using validated histological scoring systems.ConclusionsOur approach of capturing inflammatory mediator signature at different stages of UC by combining comprehensive lipidomics analysis and computational modelling could be used to classify and predict mild-moderate inflammation; however, predictive index is diminished in severe inflammation. This new technical approach could be developed to tailor drug treatments to patients with active UC, based on the mucosal lipid mediator profile.

Expression of inflammatory genes in the colon of ulcerative colitis patients varies with activity both at the mRNA and protein level

Ulcerative colitis (UC) is a chronic inflammatory disorder of the gastrointestinal tract involving aberrant activation of innate and adaptive immune responses. We aimed to study the expression profiles of the susceptibility gene Nod1 and selected pro- and anti- inflammatory cytokines during different stages of UC. 65 patients with mild, severe or remission stage of UC, and 22 normal colon mucosal biopsies from control individuals were included in the study for measuring the expression of nucleotide binding oligomerization 1(Nod1) and related pro- and anti-inflammatory cytokines using quantitative reverse transcription-PCR (qRT-PCR). mRNA expression levels were then correlated with severity of disease. In order to check their expression at the protein level, immunohistochemistry (IHC) was performed using Nod1, TNF-α, IFN-γ, IL-17, IL-23 and IL-13 antibodies. Significant increases in Nod1 expression with simultaneous increases in pro-inflammatory cytokines TNF-α, INF-γ, IL-17 and IL-23 mRNA levels were observed in patients with mild and severe ulcerative colitis versus control individuals. The expression levels reverted back towards normal levels in patients during remission. However, mRNA expression of selected anti-inflammatory cytokines such as IL-11 and IL-13 were substantially lower in patients compared with control samples when measured using qRT PCR. Levels of IL-10 however, although exhibiting a decreasing trend, did not attain significance. Our results show that with simultaneous increase in Nod1 expression, expression profiling of downstream inflammatory cytokines that are activated in UC patients, displayed different patterns according to the severity of the disease. These may be potential prognostic biomarkers for diagnosing UC patients.