Dextran Sulfate Sodium Control Research Articles

Lycium barbarum polysaccharide alleviates DSS-induced chronic ulcerative colitis by restoring intestinal barrier function and modulating gut microbiota

Purpose This study examined the protective effects and mechanism of Lycium barbarum polysaccharides (LBP) in the context of intestinal barrier function and intestinal microbiota in mice with dextran sulfate sodium (DSS)-induced chronic ulcerative colitis (UC). Methods C57BL/6J male mice were assigned to a standard normal diet without DSS (control group), a normal diet with DSS (DSS group, 2% DSS given discontinuously for 3 weeks) or a normal diet supplemented with LBP (1% dry feed weight, LBP group, 2% DSS given discontinuously for 3 weeks) for a total of 8 weeks, at which point colonic tissues and caecal contents were collected. Results LBP exerted a significant effect against colitis by increasing body weight, colon length, DAI and histopathological scores. LBP inhibited proinflammatory cytokines (IL-1β, IL-6, iNOS and TNF-α) expression, improved anti-inflammatory cytokine (IL-10) expression, promoted the expression of tight junction proteins (Occludin and ZO-1) via nuclear factor erythroid 2-related factor 2 (Nrf2) activation and decreased Claudin-2 expression to maintain the intestinal mucosal barrier. In addition, the abundances of some probiotics (Ruminococcaceae, Lactobacillus, Butyricicoccus, and Akkermansia) were decreased with DSS treatment but increased obviously with LBP treatment. And LBP reduced the abundance of conditional pathogens associated with UC (Mucispirillum and Sutterella). Furthermore, LBP improved the production of short-chain fatty acids (SCFAs), including acetic acid, propionic acid, butyric acid and isobutyric acid. Conclusion LBP can alleviate DSS-induced UC by regulating inflammatory cytokines and tight junction proteins. Moreover, LBP promotes probiotics, suppresses conditional pathogens and increases SCFAs production, showing a strong prebiotic effect.

Effect of ethyl gallate and propyl gallate on dextran sulfate sodium (DSS)-induced ulcerative colitis in C57BL/6J mice: preventive and protective.

Inflammatory bowel disease (IBD) is an idiopathic inflammatory condition of the digestive system marked by oxidative stress, leukocyte infiltration, and elevation of inflammatory mediators. In this study, we demonstrate the protective effect of ethyl gallate (EG), a phytochemical, and propyl gallate (PG), an anti-oxidant, given through normal drinking water (DW) and copper water (CW) in various combinations, which had a positive effect on the amelioration of DSS-induced ulcerative colitis in C57BL/6J mice. We successfully determined the levels of proinflammatory cytokines and anti-oxidant enzymes by ELISA, tracked oxidative/nitrosative stress (RO/NS) by in vivo imaging (IVIS) using L-012 chemiluminescent probe, disease activity index (DAI), and histopathological and morphometric analysis of colon in DSS-induced colitis in a model. The results revealed that oral administration of ethyl gallate and propyl gallate at a dose of 50mg/kg considerably reduced the severity of colitis and improved both macroscopic and microscopic clinical symptoms. The level of proinflammatory cytokines (TNF-α, IL-6, IL-1β, and IFN-γ) in colonic tissue was considerably reduced in the DSS + EG-treated and DSS + PG-treated groups, compared to the DSS alone-treated group. IVIS imaging of animals from the DSS + EG and DSS + PG-treated groups showed a highly significant decrease in RO/NS species relative to the DSS control group, with the exception of the DSS + PG/CW and DSS + EG + PG/CW-treated groups. We also observed lower levels of myeloperoxidase (MPO), nitric oxide (NO), and lipid peroxidation (LPO), and restored levels of GST and superoxide dismutase (SOD) in DSS + EG-DW/CW, DSS + PG/DW, and DSS + EG + PG/DW groups compared to DSS alone-treated group. In addition, we showed that the EG, PG, and EG + PG treatment significantly reduced the DAI score, and counteracted the body weight loss and colon shortening in mice compared to DSS alone-treated group. In this 21-day study, mice were treated daily with test substances and were challenged to DSS from day-8 to 14. Our study highlights the protective effect of ethyl gallate and propyl gallate in various combinations which, in pre-clinical animals, serve as an anti-inflammatory drug against the severe form of colitis, indicating its potential for the treatment of IBD in humans. In addition, propyl gallate was investigated for the first time in this study for its anti-colitogenic effect with normal drinking water and reduced effect with copper water.

Purification methods to reduce interference by dextran sodium sulfate with quantification of gene expression in intestinal tissue samples from a piglet model of colitis.

Dextran sodium sulfate (DSS) is commonly used to induce intestinal (i.e., colonic) inflammation in a variety of animal models. However, DSS is known to cause interference when using quantitative-real time polymerase chain reaction (qRT-PCR) methods, thereby invalidating accurate and precise measurement of tissue gene expression. Therefore, the goal of this study was to determine whether different mRNA purification methods would reduce DSS-interference. Colonic tissue samples were collected at postnatal days (PND) 27 or 28 from pigs that had not been administered DSS (Control), and two independent groups of pigs that received 1.25 g of DSS/kg of BW/d (DSS-1 and DSS-2) from PND 14 to 18. Tissue samples collected were subsequently stratified into three purification methods (i.e., 9 total treatment × method combinations), including: 1) no purification, 2) purification with lithium chloride (LiCl), or 3) purification using spin column filtration. All data were analyzed using a one-way ANOVA in the Mixed procedure of SAS. The average RNA concentrations across all treatments were between 1,300 and 1,800 μg/μL for all three in vivo groups. Although there were statistical differences among purification methods, the 260/280 and 260/230 ratios fell between acceptable limits of 2.0 to 2.1 and 2.0 to 2.2, respectively, for all treatment groups. This confirms the RNA quality was adequate and not influenced by purification method in addition to suggesting the absence of phenol, salts, and carbohydrate contamination. For pigs in the Control group that did not receive DSS, qRT-PCR Ct values of four cytokines were achieved, though these values were not altered by purification method. For pigs that had undergone DSS dosing, those tissues subjected to either no purification or purification using LiCl did not generate applicable Ct values. However, when tissues derive from DSS-treated pigs underwent spin column purification, half of the samples from DSS-1 and DSS-2 groups generated appropriate Ct estimates. Therefore, spin column purification appeared to be more effective than LiCl purification, but no method was 100% effective, so caution should be exercised when interpreting gene expression results from studies where animals are exposed to DSS-induced colitis.

Maize near-isogenic lines with enhanced flavonoids alleviated dextran sodium sulfate-induced murine colitis via modulation of the gut microbiota.

The rising incidence of inflammatory bowel disease (IBD) has necessitated the search for safe and effective novel therapeutic strategies. Dietary flavonoids exhibited antioxidant, antiproliferative, and anticarcinogenic activities in several model systems with proven abilities to reduce inflammation and oxidative stress, thus they could be promising therapeutic agents for IBD prevention/treatment. However, understanding the role of a specific class of compounds in foods that promote health is difficult because of the chemically complex food matrices. This study aimed to utilize four maize near-isogenic lines to determine the anti-colitis effects of specific classes of flavonoids, anthocyanins and/or phlobaphenes, in a whole-food matrix. Results showed that the intake of anthocyanin and phlobaphene-enriched maize diets effectively alleviated dextran sodium sulfate (DSS)-induced colitis in mice via reducing the intestinal permeability and restoring the barrier function. Anthocyanin diets were more effective in maintaining the crypt structure and muc2 protein levels and reducing inflammation. Bacterial communities of mice consuming diets enriched with anthocyanins and phlobaphenes were more similar to the healthy control compared to the DSS control group, suggesting the role of flavonoids in modulating the gut microbiota to retrieve intestinal homeostasis. Microbiota depletion rendered these compounds ineffective against colitis. Lower serum concentrations of several phenolic acids were detected in the microbiota-depleted mice, indicating that gut microbiota plays a role in flavonoid metabolism and bioavailability.

Oral gamma-cyclodextrin-encapsulated tributyrin supplementation in young pigs with experimentally induced colitis.

Disruption of intestinal integrity and barrier function due to tissue inflammation has negative implications on overall growth and well-being in young pigs. In this study, we investigated the effects of oral gamma-cyclodextrin-encapsulated tributyrin (TBCD) in young pigs experiencing dextran sodium sulfate (DSS)-induced colitis. Pigs (n = 32 boars) were weaned from the sow at postnatal day (PND) 2, allotted to treatment based on the litter of origin and body weight (BW), and reared artificially over a 26-d feeding period. Treatment groups included: 1) nutritionally adequate (control) milk replacer, no DSS (Control n = 8), 2) control milk replacer plus oral DSS (DSS, n = 7), and 3) control diet supplemented with 8.3 g of TBCD per kg of reconstituted milk replacer plus oral DSS (TBCD + DSS, n = 8). Colitis was induced by administering DSS at 1.25 g of DSS/kg BW daily in a reconstituted milk replacer from PND 14-18. Milk replacer and water were provided ad libitum throughout the 26-d study. All the data were analyzed using a one-way ANOVA using the MIXED procedure of SAS. Control and DSS pigs had similar BW throughout the study, while TBCD + DSS pigs exhibited decreased (P < 0.05) BW starting at approximately PND 15. Additionally, average daily gain (ADG) before and after initiation of DSS dosing, along with over the total study duration, was decreased (P < 0.05) in pigs receiving TBCD + DSS compared with the Control. Milk disappearance was decreased (P < 0.05) in TBCD + DSS pigs when compared with Control and DSS groups. Both the concentration and molar ratio of cecal butyrate concentrations were increased (P < 0.05) in TBCD + DSS pigs compared with the Control group. The DSS and TBCD + DSS treatments also increased (P < 0.05) butyrate concentrations in the luminal contents with the proximal colon compared with Control. TBCD + DSS and DSS pigs had increased (P < 0.05) mucosal width in the distal colon compared with Control, thereby indicating heightened intestinal inflammation. Overall, oral supplementation of encapsulated tributyrin increased the concentration of butyrate in the colon, but was unable to mitigate the negative effects of DSS-induced colitis.

Ameliorative effect of two structurally divergent hydrazide derivatives against DSS-induced colitis by targeting Nrf2 and NF-κB signaling in mice.

The environmental factors and genetic vulnerability trigger the inflammatory bowel diseases (IBDs) such as ulcerative colitis and Crohn's disease. Furthermore, the oxidative stress and inflammatory cytokines have been implicated in the aggravation of the IBDs. The aim of the present study was to investigate the effect of N-(benzylidene)-2-((2-hydroxynaphthalen-1-yl)diazenyl)benzohydrazides (NCHDH and NTHDH) compounds against the DSS-induced colitis in mice. The colitis was induced by 5% dextran sulfate sodium (DSS) dissolved in normal saline for 5days. The effect of the NCHDH and NTHDH on the behavioral, biochemical, histological, and immunohistological parameters was assessed. The NCHDH and NTHDH treatment improved the behavioral parameters such as food intake, disease activity index, and diarrhea score significantly compared to DSS control. The NCHDH and NTHDH treatments significantly increased the antioxidant enzymes, whereas oxidative stress markers were markedly reduced. Similarly, the NCHDH and NTHDH treatments significantly suppressed the activity of nitric oxide (NO), myeloperoxidase (MPO), and eosinophil peroxidase (EPO). The histological studies showed a significant reduction in inflammation, immune cell infiltration, and fibrosis in the NCHDH- and NTHDH-treated groups. The immunohistochemical results demonstrated that NCHDH and NTHDH treatments markedly increase the expression level of Nrf2, HO-1 (hemeoxygenase-1), TRX (thioredoxin reductase), and IκB compared to the DSS-induced group. In the same way, the NCHDH and NTHDH significantly reduced the NF-κB and COX-2 (cyclooxygenase-2) expression levels. The NCHDH and NTHDH treatment significantly improved the symptoms associated with colitis via inducing antioxidants and attenuating oxidative stress markers.

L-citrulline enriched fermented milk with Lactobacillus helveticus attenuates dextran sulfate sodium (DSS) induced colitis in mice

Inflammatory bowel disease (IBD) is a group of chronic inflammatory gastrointestinal diseases that causes worldwide suffering. L. helveticus is a probiotic that can enhance intestinal barrier function via alleviation of excessive inflammatory response. Citrulline, a functional amino acid, has been reported to stimulate muscle synthesis and to function with a prebiotic-like action with certain Lactobacillus strains. The aim of this study was to investigate the potential synergistic effect of combining L. helveticus and citrulline on protection against damage induced by dextran sulfate sodium (DSS) in a mouse model. 6-week-old male C57BL/6J mice were fed with DSS water and randomly divided for administering with different milk treatments: 1) plain milk (control or DSS control), 2) 1% (w/v) citrulline enriched milk (Cit_milk), 3) milk fermented with L. helveticus (LHFM) and 4) DSS+milk fermented with L. helveticus with 1% (w/v) citrulline (Cit_LHFM). The treatment effects on the survival and macroscopic and microscopic signs were examined. All treatments presented different degrees of protective effects on attenuating the damages induced by DSS. All treatments reduced the body weight loss, disease activity index (DAI), histological scores, pro-inflammatory cytokine expression (IL-6, TNF-α and IFN-γ) and production (IL-4) (all P <0.05) and the tight junction (TJ) protein (zonula occluden-1 (ZO-1) expression. LHFM and Cit_LHFM improved survival rate (both at P<0.05). Particularly, Cit_LHFM showed greater effects on protecting the damages induced by DSS, especially in ameliorating colonic permeability, TJ protein (ZO-1, occludin and claudin-1) expression and distribution as well as in reducing IL-4 and IL-17 expression (all P <0.05). Our findings suggested that the combination of and citrulline had significant synergistic effect on protecting against injury from DSS-induced colitis. Therefore, citrulline enriched L. helveticus fermented milk is suggested to be a potential therapy for treating IBD.

IL-33 Drives Expansion of Type 2 Innate Lymphoid Cells and Regulatory T Cells and Protects Mice From Severe, Acute Colitis.

The hallmarks of inflammatory bowel disease are mucosal damage and ulceration, which are known to be high-risk conditions for the development of colorectal cancer. Recently, interleukin (IL)-33 and its receptor ST2 have emerged as critical modulators in inflammatory disorders. Even though several studies highlight the IL-33/ST2 pathway as a key factor in colitis, a detailed mode of action remains elusive. Therefore, we investigated the role of IL-33 during intestinal inflammation and its potential as a novel therapeutic target in colitis. Interestingly, the expression of IL-33, but not its receptor ST2, was significantly increased in biopsies from the inflamed colon of IBD patients compared to non-inflamed colonic tissue. Accordingly, in a mouse model of Dextran Sulfate Sodium (DSS) induced colitis, the secretion of IL-33 significantly accelerated in the colon. Induction of DSS colitis in ST2-/- mice displayed an aggravated colon pathology, which suggested a favorable role of the IL 33/ST2 pathway during colitis. Indeed, injecting rmIL-33 into mice suffering from acute DSS colitis, strongly abrogated epithelial damage, pro-inflammatory cytokine secretion, and loss of barrier integrity, while it induced a strong increase of Th2 associated cytokines (IL-13/IL-5) in the colon. This effect was accompanied by the accumulation of regulatory T cells (Tregs) and type 2 innate lymphoid cells (ILC2s) in the colon. Depletion of Foxp3+ Tregs during IL-33 treatment in DSS colitis ameliorated the positive effect on the intestinal pathology. Finally, IL-33 expanded ILC2s, which were adoptively transferred to DSS treated mice, significantly reduced colonic inflammation compared to DSS control mice. In summary, our results emphasize that the IL-33/ST2 pathway plays a crucial protective role in colitis by modulating ILC2 and Treg numbers.

Fusobacterium nucleatum impedes remission of colitis in a mouse model.

The role of Fusobacterium nucleatum, often associated with intestinal diseases, in the remission of dextran sulfate sodium (DSS)-induced colitis was investigated. Female mice were divided into groups DC (DSS control) and DF (DSS+F. nucleatum). F. nucleatum (1.0×1010 cfu/mouse/day) in phosphate-buffered saline (PBS) was orally given to DF, while DC had PBS only. All mice had DSS in drinking water. In Experiment 1, mice underwent 2 inflammation phases, an in-between recovery phase and had their disease activity indices (DAI) calculated. Experiment 2 was similarly conducted, except that mice were dissected 3 days postrecovery, and had blood and colonic mucosal samples collected. In Experiment 1, DF had significantly (P<.05) higher DAI than DC, during the recovery and 2nd inflammation phases. In Experiment 2, genus Bacteroides was significantly (P<.05) higher and family Lachnospiraceae significantly lower in cecal mucosa-associated microbiota of DF than in that of DC. We concluded that F. nucleatum can impede colitis remission.

Protective Effect of Prunus mume Fermented with Mixed Lactic Acid Bacteria in Dextran Sodium Sulfate-Induced Colitis.

The fruit of Prunus mume (PM) is widely cultivated in East Asia, and it has been used as a folk medication for gastrointestinal disorders, e.g., diarrhea, stomach ache and ulceration. In this study, the pectinase-treated PM juice (PJ) was fermented with Lactobacillus strains containing fundamental organic acids and free amino acids. The PJ fermented with Lactobacillus plantarum and L. casei (FP) was investigated for its protective effect in dextran sodium sulfate (DSS)-induced colitis mice model. The administration of FP reduced lipid peroxidation and histopathological colitis symptoms, e.g., shortening of the colon length, depletion of mucin, epithelial injury and ulceration, in colonic tissues. The FP-supplemented group showed the alleviation of pro-inflammatory cytokines. Compared with the DSS control group, the supplementation of FP significantly reduced the levels of serum interferon-γ (IFN-γ), interleukin (IL)-1β, IL-6, IL-12 and IL-17 as well as colonic tumor necrosis factor-α, IFN-γ, IL-12 and IL-17. Furthermore, the DSS-induced TUNEL-positive area was significantly reduced by the FP supplementation. These results show that the supplementation of FP fermented with mixed lactic acid bacteria, L. plantarum and L. casei, elucidated the protective effect in DSS-induced colitis mice. Hence, this study suggests that FP can be utilized as a natural therapeutic agent for colitis and intestinal inflammation.

Intrarectally administered polaprezinc attenuates the development of dextran sodium sulfate-induced ulcerative colitis in mice.

Polaprezinc (PZ), a chelate of zinc and L-carnosine, has been widely used in the treatment of gastric ulcers since 1994. In recent years, researchers have found PZ to have a beneficial effect on various experimentally induced models of colitis in mice. In the present study, 6% dextran sodium sulfate (DSS) was used to induce a model of ulcerative colitis (UC) in Institute of Cancer Research mice. The therapeutic effect and mechanism of PZ action in a model of UC was studied in order to provide an experimental basis for the clinical application of PZ in UC treatment. The effect of PZ on UC was evaluated in five groups of mice: A vehicle control only group, a DSS model control group (DSS, 6%), a validated treatment control group (DSS 6% + Mesalamine), a low-dose PZ treatment group (DSS 6% + PZ 60 mg/kg) and a high-dose PZ group (DSS 6% + PZ 120 mg/kg). After the animals were sacrificed, blood was collected and the serum levels of NF-κB and tumor necrosis factor-α (TNF-α) were measured. Changes in histology were observed by light microscopy. The protein levels of AKT, phosphorylated AKT and heat shock protein 70 (HSP70) were determined by western blot analysis. The results suggested that PZ reduced the DSS-induced increase in the inflammatory proteins TNF-α and NF-κB in the UC model. The high-dose of PZ also increased the HSP70 protein level, inhibited AKT phosphorylation in a DSS-induced UC animal model, and decreased white blood cell and neutrophil % counts compared to levels in an untreated DSS control group. Histopathology indicated that the mice of the DSS model group had irregular colonic villi, a large number of inflammatory cells and mucosal damage, whereas mice of the group treated with PZ had small intestinal villus morphology and their villi showed signs of recovery from the damage of UC. The results of the present study indicated that PZ significantly alleviates DSS-induced UC in mice, relieves diarrhea, and inhibits the phosphorylation of inflammatory factors and the inflammatory AKT signaling pathway.

TNF targeting small molecule attenuates colonic inflammation in acute DSS-induced colitis mice

Abstract Background Inflammatory bowel disease (IBD) is a chronic and multifactorial inflammatory disorder which is comprising Crohn’s disease (CD) and ulcerative colitis (UC). Although the incidence of IBD has steady increase in worldwide, the conventional drug therapies with 5-aminosalicylates, corticosteroids, and immunosuppressants are insufficient to maintain remission and can occur side effects. Therefore, other class of drug therapies using specific cytokine inhibitor have recently been focused. In this study, we aimed to investigate that anti-inflammatory effects of TNF targeting small molecule in dextran sodium sulfate (DSS)-induced colitis model. Methods C57BL/6 (21–23g) mice were administered 2–2.5% DSS in the drinking water for 5 days to induce acute colitis. Different doses of IA-14069 (20, 40, 80, 160 mg/kg) were orally treated daily for 5–12 days and the mice were sacrificed for the experiments. Normal or DSS control group was given drinking water only or DSS only. Disease severity, body weight change, and colon length were measured, and serum levels of TNF, IFN-g, IL-6, IL-17 and IL-5 were analyzed by ELISA. Colonic epithelial damage was observed by H&amp;E staining of colon tissue. Results Compared with normal group, oral administration of 80 mg/kg IA-14069 significantly inhibited the severity of colitis, loss of body weight, and colon shortening of the DSS-induced colitis mice. Moreover, mucosal damages and production of the proinflammatory cytokines were markedly suppressed by treatment of IA-14069. Conclusion These findings suggest that IA-14069, a TNF-targeting small molecule, exerts a suppressive effect on DSS-induced colitis and may be used as an IBD therapeutic agent.

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

Lactobacillus fermentum species ameliorate dextran sulfate sodium-induced colitis by regulating the immune response and altering gut microbiota

ABSTRACT We evaluated immunometabolic functions of novel Lactobacillus fermentum strains (KBL374 and KBL375) isolated from feces of healthy Koreans. The levels of inflammatory cytokines, such as interleukin (IL)-2, interferon-γ, IL-4, IL-13, and IL-17A, were decreased, and that of the anti-inflammatory cytokine IL-10 was increased, in human peripheral blood mononuclear cells (PBMCs) treated with the L. fermentum KBL374 or KBL375 strain. When these strains were orally administered to mice with dextran sulfate sodium (DSS)-induced colitis, both L. fermentum KBL374 and KBL375 showed beneficial effects on body weight, disease activity index score, colon length, cecal weight, and histological scores. Furthermore, both L. fermentum KBL374 and KBL375 modulated the innate immune response by improving gut barrier function and reducing leukocyte infiltration. Consistent with the PBMC data, both L. fermentum KBL374- and KBL375-treated DSS mice demonstrated decreased Th1-, Th2-, and Th17-related cytokine levels and increased IL-10 in the colon compared with the DSS control mice. Administration of L. fermentum KBL374 or KBL375 to mice increased the CD4+CD25+Foxp3+Treg cell population in mesenteric lymph nodes. Additionally, L. fermentum KBL374 or KBL375 administration reshaped and increased the diversity of the gut microbiota. In particular, L. fermentum KBL375 increased the abundance of beneficial microorganisms, such as Lactobacillus spp. and Akkermansia spp. Both L. fermentum KBL374 and KBL375 may alleviate inflammatory diseases, such as inflammatory bowel disease, in the gut by regulating immune responses and altering the composition of gut microbiota.

Protective Effects of Pioglitazone against Dextran Sodium Sulphate-Induced Colitis in Rats.

The aim of the present study is to investigate the preventive effect of pioglitazone on colitis induced with dextran sulfate sodium (DSS) in rats. Twenty-four male Sprague-Dawley rats weighing 180-200 g were randomized into four groups. Rats of the 1st group received only saline and served as normal group. Colitis was induced in the remaining 3 groups by 1.5% DSS administered in drinking water for 8 days. Group 2 received only saline p.o. for 30 days and served as DSS control group. Groups 3 and 4 received pioglitazone (PIO; 10 mg/kg/day, p.o.) and sulfasalazine (SUL; 300 mg/kg) respectively for 30 days. All animals were sacrificed 24 h after the last treatment. Serum levels of interleukin-2 (IL-2), interleukin-6 (IL-6) and interleukin-17 (IL-17) were measured. Levels of tumor necrosis factor-α (TNF-α), reduced glutathione (GSH) and malondialdehyde (MDA), were measured in colon tissue. Significant elevation in GSH levels and reduction in MDA and TNF-α levels in colon tissue were observed in pioglitazone and sulfasalazine treated group when compared with the DSS control group. Significant elevation in serum IL-2 and reduction in serum IL-6 and IL-17 were observed when compared with the DSS control group. Pioglitazone reduced DSS-induced colitis possibly via reduction of MDA, TNF-α, IL-6 and IL-17 levels.

P074 Human amnion epithelial cells reduce intestinal inflammation in a dextran sulfate sodium-induced murine model of acute colitis

Inflammatory bowel disease encompasses a group of conditions characterised by inflammation and ulceration of the colon resulting in diarrhoea, haematochezia and weight loss. This often requires hospitalisation, corticosteroids and long-term immunosuppressants. However, these treatments are associated with numerous side effects and a subset of patients remain refractory to treatment. Stem cells, such as the pluripotent human amnion epithelial cells (hAECs), have been explored as a potential therapy. Ethically sourced from placentas, hAECs possess anti-inflammatory properties and no known safety issues. We examined the efficacy of hAECs in a dextran sulfate sodium (DSS) induced murine model of acute colitis. C57BL/6J mice received 2% DSS (w/v) in their normal drinking water for 7 days. At Day 2, Group 1 (n = 17) received a tail vein of 2 × 106 hAECs. At Day 4, Group 2 (n = 17) received an injection of 2 × 106 hAECs. DSS control mice (n = 17) were untreated. Mice were monitored daily and culled at Day 8. Colitis severity was scored histologically using H&E stained colon sections. Colonic neutrophil infiltration was determined by myeloperoxidase (MPO) activity assay. IL-1β, TNF-α, IL-6, IFN-γ and IL-10 were measured through enzyme-linked immunosorbent assays. Colon sections were stained with anti-F/480 antibody to identify macrophages. Untreated DSS mice lost significantly more body weight (−17.4%) compared with hAEC treated mice (Group 1: −5.6% (p < 0.0001), Group 2: −8.8% (p < 0.0001)). Treatment prevented colon shortening in Group 1 (6.7 cm p < 0.001) and Group 2 (6.3 cm, p < 0.01) compared with controls (5.3 cm). Similarly, hAEC treated mice had significantly lower histological severity scores (Group 1: p < 0.0001, Group 2: p < 0.001). MPO activity was almost halved in Groups 1 (p < 0.0001) and 2 (p < 0.0001). Treatment also significantly reduced concentrations of pro-inflammatory cytokines (IL-1β, TNF-α, IL-6, IFN-γ) whilst up-regulating the anti-inflammatory cytokine IL-10. hAEC treatment significantly reduced intestinal macrophages at Day 2 (p < 0.01) but not Day 4. Apart from a lower histological severity score in Group 1, there were no others differences comparing Group 1 and 2. hAECs exert a protective effect in colitis and reduce the severity of disease, by reducing the level of neutrophil and macrophage infiltration into the colon and decreasing concentrations of pro-inflammatory cytokines and increasing anti-inflammatory cytokine. The therapeutic potential of hAECs in IBD is promising and further research into their optimal delivery and mechanisms of action is warranted.

Orally administered sodium 4-phenylbutyrate suppresses the development of dextran sulfate sodium-induced colitis in mice.

Sodium 4-phenylbutyrate (PBA) exerts therapeutic effects in a wide range of pathologies. A previous study by the present authors revealed that intraperitoneal administration of PBA suppresses the onset of dextran sulfate sodium (DSS)-induced colitis in mice. In the present study, the effects of orally administered PBA are investigated, as this route of administration is more clinically relevant. The therapeutic efficacy of PBA (10 mg/12 h) in mice with experimental colitis was assessed based on the disease activity index, production of inflammatory cytokines, colon length and histopathological investigations. The results of the present study demonstrated a significantly higher survival rate in the PBA-treated group compared with the PBA-untreated (DSS control) group (P=0.0156). PBA treatment improved pathological indices of experimental colitis (P<0.05). Furthermore, the oral administration of PBA significantly inhibited the DSS-induced shortening of the colon (P<0.05) and overproduction of interleukin (IL)-1β and IL-6 (both P<0.05) as measured in colonic lavage fluids. A marked attenuation of the DSS-induced overproduction of tumor necrosis factor was also observed. For histopathological analysis, a marked decrease in mature goblet cells and increase in enlarged nuclei of the absorptive cells was observed in colon lesions of DSS control mice as compared with normal untreated mice. However, in the PBA-treated mice, no such lesions were observed and the mucosa resembled that of DSS-untreated mice. The results of the present study, combined with those results of a previous study, suggest that oral and intraperitoneal administration of PBA have similar preventative effects on DSS-induced colitis, achieved by suppressing its pathogenesis.

Abstract B40: Effect of Sasa quelpaertensis leaf extracts on the gut microbiota in dextran sodium sulfate-induced colitis in mice

Abstract Inflammatory bowel diseases (IBD), including Crohn’s disease and ulcerative colitis, are chronic inflammatory disorders of gastrointestinal tract. Although the etiology of IBD is not clear, it may be associated with genetics and lifestyle factors, diet and gut microbiome. In particular, the imbalance among the immune system and gut microbiota and diet may play a key role in developing IBD. Sasa quelpaertensis leaves have shown anti-inflammatory and anticarcinogenic effects, although the effect of these leaves on gut microbiota in colitis remains unclear. The present study hypothesized that Sasa quelpaertensis leaves extract (SQE) exerts a protective effect against a dextran sulfate sodium (DSS)-induced colitis in mice through modulating gut microbiota. SQE (100 mg/kg or 300 mg/kg b.w.) were pre-treated to mice by gavage two weeks before inflammation which was induced by 2.5% DSS in drinking water for two cycles of each 7 days and tap water for 7 days was provided between DSS treatment. Disease activity index (DAI), colon length were measured and microbiota were analyzed using pyrosequencing in fecal samples. SQE attenuated the severity of DSS-induced colitis. In addition, SQE group had more Firmicutes in phylum level compared to DSS group, Bacteroidetes/Firmicutes ratio of DSS group was significantly higher than control and SQE groups. Furthermore, major microbiota compositions were different in class, family, genus, and species level among control, DSS control, and SQE group. Estimator of community richness, the OTUs, in DSS group tended to lower compared with SQE groups and estimator of evenness and diversity, Chao-1, ACE, Shannon, NP Simpson, were significantly lower in DSS group compared with SQE groups (p&amp;lt;0.05). Our results suggest that SQE affects microbiome composition and number change on DSS-induced colitis mouse model, these change in the microbiome may be beneficial and can be an effective strategy for prevention of IBD (This work was conducted with the support of the Cooperative Research Program for Agriculture Science &amp; Technology Development (Project No. PJ009777) Rural Development Administration, Republic of Korea, and the Brain Korea 21 Plus (Project No. 22A20130012143). Citation Format: Yiseul Yeom, Yuri Kim. Effect of Sasa quelpaertensis leaf extracts on the gut microbiota in dextran sodium sulfate-induced colitis in mice. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B40.

Intestinal anti-inflammatory activity of the seeds of Raphanus sativus L. in experimental ulcerative colitis models

Ethnopharmacological relevanceWater extract of Raphanus sativus L. (RSL) seeds was traditionally used to treat digestive inflammatory complaints in Korean culture. RSL seeds exerted antioxidant, anti-inflammatory, and anti-septic functions, suggesting their pharmacological potential for the treatment of inflammatory pathologies associated with oxidative stress such as inflammatory bowel disease. Aim of this studyWe evaluated the intestinal anti-inflammatory effects of RSL seed water extract (RWE) in experimental rat models of trinitrobenzenesulphonic acid (TNBS)- or dextran sodium sulfate (DSS)-induced colitis. Materials and methodsRWE was characterized by determining the content of sinapic acid as a reference material and then assayed in the DSS and TNBS models of rat colitis. Male Sprague–Dawley rats were divided into 10 groups (n=7/group): non-colitic control, DSS or TNBS control, DSS colitis groups treated with RWE (100mg/kg) or mesalazine (25mg/kg), and TNBS colitis groups treated with various doses (10, 40, 70, and 100mg/kg) of RWE or mesalazine (25mg/kg). RWE or mesalazine treatment started the same day of colitis induction and rats were sacrificed 24h after the last treatment followed by histological and biochemical analyses. ResultsOral administration with RWE suppressed intestinal inflammatory damages in both DSS- and TNBS-induced colitic rats. The treatment with 100mg/kg RWE recovered intestinal damages caused by TNBS or DSS to levels similar to that of mesalazine, decreasing the activity of myeloperoxidase activity and the secretion of tumor necrosis factor (TNF)-α and interleukin (IL)-1β. RWE treatment inhibited malondialdehyde production and glutathione reduction in colon of colitis rats. The administration of RWE at dose of 100mg/kg also suppressed the TNBS- or DSS-stimulated expression of TNF-α, IL-1β, monocyte chemotactic protein-1, inducible nitric oxide, and intercellular adhesion molecule-1. Furthermore, RWE inhibited p38 kinase and DNA–nuclear factor-κB binding activities, both of which were stimulated in the colitic rats. ConclusionsThe current findings show that RWE ameliorates intestinal oxidative and inflammatory damages in DSS and TNBS models of rat colitis, suggesting its beneficial use for the treatment of intestinal inflammatory disorders.

Jumihaidokuto effectively inhibits colon inflammation and apoptosis in mice with acute colitis

Jumihaidokuto, a Japanese kampo medicine, is prescribed in Japan for its anti-inflammatory activity. Here we have examined its beneficial effects against acute colitis induced by dextran sulfate sodium (DSS) in mice. We have used C57BL/6 female mice, divided into two groups and received 3% DSS in drinking water during the experimental period (8days). Treatment group mice received 1g/kg/day dose of Jumihaidokuto orally whereas DSS control group received equal volume of distilled water. Normal control group mice received plain drinking water. Jumihaidokuto treatment attenuated the colitis symptoms along with suppression of various inflammatory marker proteins such as IL-1β, IL-2Rα, IL-4, CTGF and RAGE. It has also down-regulated the oxidative stress and apoptotic signaling in the colons of mice with colitis. The present study has confirmed the beneficial effects of Jumihaidokuto on DSS induced acute colitis in mice and suggests that it can be a potential agent for the treatment of colitis.