Inflammatory bowel disease(IBD)is a complex relapsing inflammatory disease in the gut and is driven by complicated host-gut microbiome interactions.Gut commensals have shown different functions in IBD prevention and t...Inflammatory bowel disease(IBD)is a complex relapsing inflammatory disease in the gut and is driven by complicated host-gut microbiome interactions.Gut commensals have shown different functions in IBD prevention and treatment.To gain a mechanistic understanding of how different commensals affect intestinal inflammation,we compared the protective effects of 6 probiotics(belonging to the genera Akkermansia,Bifidobacterium,Clostridium,and Enterococcus)on dextran sulfate sodium(DSS)-induced colitis in mice with or without gut microbiota.Anti-inflammatory properties(ratio of interleukin(IL)-10 and IL-12)of these strains were also evaluated in an in vitro mesenteric lymph nodes(MLN)co-culture system.Results showed that 4 probiotics(belonging to the species Bifidobacterium breve,Bifidobacterium bifidum,and Enterococcus faecalis)can alleviate colitis in normal mice.The probiotic strains differed in regulating the intestinal microbiota,cytokines(IL-10,IL-1βand interferon(IFN)-γ),and tight junction function(Zonulin-1 and Occludin).By constrast,Akkermansia muciniphila AH39 and Clostridium butyricum FHuNHHMY49T1 were not protective.Interestingly,B.breve JSNJJNM2 with high anti-inflammatory potential in the MLN model could relieve colitis symptoms in antibiotic cocktail(Abx)-treated mice.Meanwhile,E.faecalis FJSWX25M1induced low levels of cytokines in vitro and showed no beneficial effects.Therefore,we provided insight into the clinical application of probiotics in IBD treatment.展开更多
AIM To assess the effect of sodium selenite on the severity of dextran sulfate sodium(DSS)-induced colitis in C57BL/6 mice.METHODS Mice were randomly divided into four groups(n = 10/group): normal group, selenium(Se) ...AIM To assess the effect of sodium selenite on the severity of dextran sulfate sodium(DSS)-induced colitis in C57BL/6 mice.METHODS Mice were randomly divided into four groups(n = 10/group): normal group, selenium(Se) group, chronic colitis group, and Se + chronic colitis group. The mice were sacrificed on day 26. Survival rates, clinical symptoms, colon length, and histological changes were determined. The percentages and absolute numbers of immune system cells in the lamina propria lymphocytes(LPL) of the colon, the expression of m RNA in colon tissue, and the concentrations of Th1, Th17, and Treg cytokines in LPL from the large intestine, were measured.RESULTS Se significantly ameliorated the symptoms of colitis and histological injury(P < 0.05 each), increasing the proportions of neutrophils and CD4+ CD25+ T cells(P < 0.05 each) and decreasing the proportions of γδT cells, CD4+, CD4+CD44+, and CD4+ CD69+ T cells in LPL(P < 0.05 each). Moreover, Se reduced the expression of IL-6, IFN-γ, IL-17 A, IL-21, T-bet, and RORγt(P < 0.05 each), but enhanced the expression of IL-10 and Foxp3(P < 0.05 each). CONCLUSION These results suggest that Se protects against DSSinduced chronic colitis perhaps by increasing the number of CD4(+)CD25(+) Tregs that suppress the secretion of proinflammatory cytokines and populations of Th1, Th17, and γδT cells.展开更多
BACKGROUND Intestinal micro-ecological imbalances impair the intestinal barrier and induce intestinal inflammation,for example,ulcerative colitis(UC).According to the latest research,abnormalities in intestinal microb...BACKGROUND Intestinal micro-ecological imbalances impair the intestinal barrier and induce intestinal inflammation,for example,ulcerative colitis(UC).According to the latest research,abnormalities in intestinal microbiota structure and their metabolites play a dominant role in UC progression;in addition,they could affect the mucus barrier based on different factors.Although numerous studies have confirmed the important role of intestinal microbiota in UC pathogenesis,the intricate connection between microbiota and metabolites and mucus barrier in UC occurrence remains unclear,and correlation analyses of differential microbiota and their metabolites under UC are relatively scarce.AIM To reveal the differential intestinal microbiota and metabolites in UC pathogenesis and explore more sensitive biomarker compositions.METHODS We used the antibiotic combination method to establish intestinal pseudo-aseptic mice;afterward,dextran sulfate sodium(DSS)was applied to establish an acute experimental colitis mice model.Colitis severity,assessed based on disease activity index,colorectal length,colorectal wet weight,and histological lesions,and mucus-related staining(mucopolysaccharide alcian blue and immunofluorescence of mucin),was compared between the pseudo-aseptic and bacterial colitis mice.Finally,differential intestinal microbiota,metabolites,and their association and correlations,were analyzed by 16s rDNA sequencing in combination with non-targeted metabolomics,through gas chromatography-mass spectrometry.RESULTS Compared with the pseudo-aseptic mice,intestinal bacteria positive mice were more severely ill and their intestinal mucus loss was more pronounced in DSS-induced colitis(P<0.05),suggesting that different microbiota and metabolites could cause the different degrees of colitis.Subsequently,we observed that in addition to Klebsiella,and Bacteroides,which were widely associated with colitis,Candidatus Stoquefichus,Anaerobiospirillum,Muribaculum,and Negativibacillus may be involved in protection against colitis.Furthermore,differential metabolites of the microbiota were mainly enriched in the synthesis-related pathways of key structural sequences of mucin.In combination with the mucin-related staining and immunofluorescence results,the findings indicate that the differential microbiota and their metabolites potentially regulate the composition and function of mucus under colitis.CONCLUSION Microbiota and their metabolites are major factors regulating the composition and function of mucus,in turn influencing the function and structure of intestinal mucus barrier under colitis.The different microbiota and metabolites identified in the present study could be novel biomarkers for colitis.展开更多
Ulcerative colitis (UC) is a lifelong illness with profound emotional and social impacts, and could cause serious damage to large intestine, especially in colon. However, the pathogenesis of UC remained unclear. The...Ulcerative colitis (UC) is a lifelong illness with profound emotional and social impacts, and could cause serious damage to large intestine, especially in colon. However, the pathogenesis of UC remained unclear. The present study attempts to find out the role of matrix metalloproteinases-7 (MMP-7) and lysozyme in the pathogenesis of UC through a mice model induced by dextran sulfate sodium (DSS). The UC model was evaluated both by disease activity index (DAI) and the intestinal histopathology. The results show that there is a high correlation between the DAI score and the pathological changes of colon. Interleukin-6 (IL-6) serum levels and large intestinal fluids levels in UC mice are always higher than that of the control groups, which might be associated with the degree of the inflammation damage in the colon. The change tendency of the MMP-7 mRNA and protein expressions are both up-regulated firstly and then down-regulated from 1 to 5 d in the colon, but only the MMP-7 protein is up-regulated at 7 d again. The up-regulated MMP-7 levels in the early stage of UC may play a protective role through the activated defensins, while the down-regulated levels in the mid-later stage of UC may be connected with the severe lesions in the colon. However, the up-regulated MMP-7 levels in the later stage of UC in the colon may also contribute to the tissue repair or be served as a marker to CRC (colorectal cancer). The distribution of lysozyme protein indicates that there may be Paneth-like cells in the colon. Both the changes of MMP-7 and lysozyme in the small intestine may play a protective role for the safe environment of the whole gut, especially to the colon of UC.展开更多
Background: Disrupted Ca2+ homeostasis contributes to the development of colonic dysmotility in ulcerative colitis (UC), but the underlying mechanisms are unknown. This study aimed to examine the alteration of col...Background: Disrupted Ca2+ homeostasis contributes to the development of colonic dysmotility in ulcerative colitis (UC), but the underlying mechanisms are unknown. This study aimed to examine the alteration of colonic smooth muscle (SM) Ca2+ signaling and Ca2+ handling proteins in a rat model of dextran sulfate sodium (DSS)-induced UC. Methods: Male Sprague-Dawley rats were randomly divided into control (n = 18) and DSS (n = 17) groups. Acute colitis was induced by 5% DSS in the drinking water for 7 days. Contractility of colonic SM strips (controls, n = 8 and DSS, n = 7) was measured in an organ bath. Cytosolic resting Ca2+ levels (n = 3 in each group) and Ca2+ transients (n = 3 in each group) were measured in single colonic SM cells. Ca2+ handling protein expression was determined by Western blotting (n = 4 in each group). Differences between control and DSS groups were analyzed by a two-sample independent t-test. Results: Average tension and amplitude of spontaneous contractions of colonic muscle strips were significantly enhanced in DSS-treated rats compared with controls (1.25 ± 0.08 g vs. 0.96 - 0.05 g, P = 0.007; and 2.67 - 0.62 g vs. 0.52 ±0.10 g, P= 0.013). Average tensions of carbachol-evoked contractions were much weaker in the DSS group (1.08 ±0.10 g vs. 1.80 ±0.19 g, P = 0.006). Spontaneous Ca2+ transients were observed in more SM cells from DSS-treated rats (15/30 cells) than from controls (5/36 cells). Peak caffeine-induced intracellular Ca2+ release was lower in SM cells of DSS-treated rats than controls (0.413 ±0.046 vs. 0.548 ±0.041, P = 0.033). Finally, several Ca2+ handling proteins in colonic SM were altered by DSS treatment, including sarcoplasmic reticulum calcium-transporting ATPase 2a downregulation and phospholamban and inositol 1,4,5-trisphosphate receptor 1 upregulation. Conclusions: Impaired intracellular Ca2+ signaling of colonic SM, caused by alteration of Ca2+ handing proteins, contribute to colonic dysmotility in DSS-induced UC.展开更多
BACKGROUND Sulfasalazine has been used as a standard-of-care in ulcerative colitis for decades,however,it results in severe adverse symptoms,such as hepatotoxicity,blood disorders,male infertility,and hypospermia.Acco...BACKGROUND Sulfasalazine has been used as a standard-of-care in ulcerative colitis for decades,however,it results in severe adverse symptoms,such as hepatotoxicity,blood disorders,male infertility,and hypospermia.Accordingly,the new treatment strategy has to enhance pharmacological efficacy and stimultaneously minimize side effects.AIM To compare the anti-inflammatory action of sulfasalazine alone or in combination with herbal medicine for ulcerative colitis in a dextran sodium sulfate(DSS)-induced colitis mouse model.METHODS To induce ulcerative colitis,mice received 5%DSS in drinking water for 7 d.Animals were divided into five groups(n=9 each)for use as normal(non-DSS),DSS controls,DSS+sulfasalazine(30 mg/kg)-treatment experimentals,DSS+sulfasalazine(60 mg/kg)-treatment experimentals,DSS+sulfasalazine(30 mg/kg)+Citrus unshiu peel and Bupleuri radix mixture(30 mg/kg)(SCPB)-treatment experimentals.RESULTS The SCPB treatment showed an outstanding effectiveness in counteracting the ulcerative colitis,as evidenced by reduction in body weight,improvement in crypt morphology,increase in antioxidant defenses,down-regulation of proinflammatory proteins and cytokines,and inhibition of proteins related to apoptosis.CONCLUSIONSCPB may represent a promising alternative therapeutic against ulcerative colitis,without inducing adverse effects.展开更多
Objective:To investigate the effect of total alkaloids of Sophora alopecuroides(TASA) on dextran sulfate sodium(DSS)-induced colitis in mice.Methods:Chronic experimental colitis was induced by administration of ...Objective:To investigate the effect of total alkaloids of Sophora alopecuroides(TASA) on dextran sulfate sodium(DSS)-induced colitis in mice.Methods:Chronic experimental colitis was induced by administration of 4 cycles of 4%DSS.Fifty mice were randomly distributed into 4 groups(normal,DSS,DSS/high-dose TASA, and DSS/low-dose TASA groups) by a random number table with body weight stratification.Mice in the normal group(n=11) and DSS-induced colitis control group(n=15) received control treatment of 20 mL/kg distilled water; DSS plus TASA high- and low-dose groups(n=12 each) were treated with TASA solution(20 mL/kg) at the doses of 60 mg/kg and 30 mg/kg,respectively.The severity of colitis was assessed on the basis of clinical signs, colon length,and histology scores.Moreover,secretory immunoglobulin A(slgA) and haptoglobin(HP) were analyzed by enzyme linked immunosorbent assay;intercellular adhesion molecule 1(ICAM-1) and macrophage-migration inhibitory factor(MIF) gene expressions were analyzed by quantitative reverse transcriptase realtime polymerase chain reaction(qRT-PCR) using SYBA greenⅠ;and nuclear factorκB(NF-κB) expression and activation and p65 interaction with the promoter of ICAM-1 gene were assessed by Western blotting and chromatin immunoprecipitation assay.Results:TASA administration significantly attenuated the damage and substantially reduced HP elevation and maintained the level of cecum slgA.TASA inhibited the ICAM-1 gene expression and had no effect on MIF gene expression.Also,TASA was able to reduce phospho-lκBα(p-lκBα) protein expression;however,it had no effect on the activation of IκB kinaseα(IKKα) and inhibitor of NF-κBα(IκBα).Moreover,TASA inhibited the p65 recruitment to the ICAM-1 gene promoter.Conclusions:TASA had a protective effect on DSS-induced colitis.Such effect may be associated with its inhibition of NF-κB activation and blockade of NF-κB-regulated transcription activation of proinflammatory mediator gene.展开更多
The role of interleukin 25 (IL-25) in a number of human diseases still has not been extensively studied, here we attempt to evaluate the role of recombinant IL-25 (rIL-25) in the development of dextran sulfate sod...The role of interleukin 25 (IL-25) in a number of human diseases still has not been extensively studied, here we attempt to evaluate the role of recombinant IL-25 (rIL-25) in the development of dextran sulfate sodium (DSS)- induced experimental colitis. Acute colitis was induced in female C57BL/6 mice by oral administration of 2.5% DSS in drinking water ad libitum. At the same time as the start of DSS exposure, mice were injected intraperitoneally with 0.4 +tg of rIL-25 or PBS. Then disease activity index (DAI), histological changes and survival rate were observed. The levels of IL-17, IL-23, and TGF-β1 in colon tissues were determined by ELISA, and the production of IL-17 by CD4+/CD8+ T cells was detected by intracellular flow cytometry. In contrast to the DSS treated mice, DSS + rIL-25 treated mice displayed a lower DAI, limited histological changes and prolonged survival. The levels of IL-23 and TGF-β1 were significantly elevated in the DSS + rIL-25 treated mice compared to the DSS treated mice. There was no significant difference in the production of IL-17 in colon tissues and CD4+/CD8+ T cells between the DSS + rIL-25 treated mice and DSS treated mice. Our findings suggest the role of IL-25 in inhibiting development and progression of acute colitis in DSS-induced mouse colitis model. Cellular & Molecular Immunology. 2008;5(6):425-431.展开更多
Background It has been reported that CD8+ regulatory cells could be induced upon oral tolerance. The purpose of this study was to investigate the changes of CD8a+ T cells in dextran sulfate sodium (DSS)-induced co...Background It has been reported that CD8+ regulatory cells could be induced upon oral tolerance. The purpose of this study was to investigate the changes of CD8a+ T cells in dextran sulfate sodium (DSS)-induced colitis mice pretreated by oral immune regulation. Methods The effects of five low oral doses of colitis-extracted proteins (CEP) on colitis were evaluated by clinical manifestation and histological lesions. The percentages of CD8a+ T cells gating on CD3+ T cells were evaluated in the gut-associated lymphoid tissues (GALT) and the spleens by flow cytometry. Differences between the two groups were compared by Student's t test or Mann-Whitney U test. Results Compared to bovine serum albumin (BSA)-fed control mice, administration of CEP resulted in marked alleviation of colitis. The proportion of CDSa+ T ceils, not only in intraepithelial lymphocytes (IELs) and lamina propria lymphocytes (LPLs) of the large intestine (LI) but also in spleen from CEP-fed colitis mice, was significantly higher than that from BSA-fed colitis mice (LI-IELs: (71.5±5.4)% vs. (60.1±4.3)%, P 〈0.01; LI-LPLs: (60.7±5.2)% vs. (51.9±4.7)%, P 〈0.01; spleen: (24.1±3.6)% vs. (20.3±4.1)%, P 〈0.05; n=8). Mucosal repair in repair-period mice five days after termination of DSS treatment was also accompanied by an increase of CD8a+ T cells in large intestinal mucosal lymphocytes (LI-IELs: (72.1±3.7)% vs. (61.5±4.5)%, P 〈0.01; LI-LPLs: (62.1±5.7)% vs. (52.7±3.6)%, P 〈0.01; n=8). The proportion of CD3+ T cells increased in Peyer's patches (PPs) and decreased in mesenteric lymph nodes (MLNs) from colitis mice compared to untreated mice, whereas the change pattern of CD3+T cells in PPs and MLNs from CEP-fed colitis mice was just on the contrary. Conclusion Improvement of DSS-induced colitis resulted from oral immune regulation is associated with an increase in CD8a+T cells in spleen and large intestinal mucosa.展开更多
Inflammatory bowel diseases(IBD),including Crohn's disease and ulcerative colitis,are complex diseases that result from the chronic dysregulated immune response in the gastrointestinal tract. The exact etiology is...Inflammatory bowel diseases(IBD),including Crohn's disease and ulcerative colitis,are complex diseases that result from the chronic dysregulated immune response in the gastrointestinal tract. The exact etiology is not fully understood,but it is accepted that it occurs when an inappropriate aggressive inflammatory respon-se in a genetically susceptible host due to inciting environmental factors occurs. To investigate the path-ogenesis and etiology of human IBD,various animal models of IBD have been developed that provided indispensable insights into the histopathological and morphological changes as well as factors associated with the pathogenesis of IBD and evaluation of therapeutic options in the last few decades. The most widely used experimental model employs dextran sodium sulfate(DSS) to induce epithelial damage. The DSS colitis model in IBD research has advantages over other various chemically induced experimental models due to its rapidity,simplicity,reproducibility and controllability. In this manuscript,we review the newer publicized advances of research in murine colitis models that focus upon the disruption of the barrier function of the intestine,effects of mucin on the development of colitis,alterations found in microbial balance and resultant changes in the metabolome specifically in the DSS colitis murine model and its relation to the pathogenesis of IBD.展开更多
Objective Patients with chronic ulcerative colitis(UC)often have mental symptoms such as depression and anxiety,and stress can lead to gastrointestinal diseases.However,the correlation between mental stress and UC is ...Objective Patients with chronic ulcerative colitis(UC)often have mental symptoms such as depression and anxiety,and stress can lead to gastrointestinal diseases.However,the correlation between mental stress and UC is unclear.In this paper,chronic unpredictable mild stress(CUMS)was utilized to evaluate the involvement of mental factors in the pathogenesis of UC.Methods The CUMS model was used to evaluate the direct/indirect involvement of mental factors in the pathogenesis of UC.The behavior was evaluated by the open field,forced swimming,and tail suspension tests.Body weight,the disease activity index(DAI)score,colon length,and HE staining of colon tissue were used to evaluate the action of CUMS and fluoxetine.Results The results showed that weight loss and the DAI score increased in CUMS mice,but they had no meaningful effect on colon length and morphological structure of colon tissue.However,CUMS aggravated dextran sulfate sodium(DSS)-induced colon length shortening and colon morphological structure damage.Fluoxetine significantly improved the DAI score,shortened colon length,and damaged morphology and structure of the colons induced by CUMS combined with DSS in mice.Fluoxetine also decreased the level of IL-6 in the serum and the TNF-αand IFN-γlevels of colon tissue.Fluoxetine simultaneously improved behavioral abnormalities induced by CUMS combined with DSS in mice.Conclusion CUMS aggravated the UC symptoms induced by DSS,and fluoxetine could improve the UC symptoms due to its improvement in the inflammatory level and behavioral abnormalities.展开更多
Background Diarrhea is a common clinical feature of ulcerative colitis resulting from unbalanced intestinal fluid and salt absorption and secretion.The Cl-/HCO3-exchanger SLC26A3 is strongly expressed in the mid-dista...Background Diarrhea is a common clinical feature of ulcerative colitis resulting from unbalanced intestinal fluid and salt absorption and secretion.The Cl-/HCO3-exchanger SLC26A3 is strongly expressed in the mid-distal colon and plays an essential role in colonic Cl-absorption and HCO3-secretion.Sic26a3 expression is up-regulated by lysophosphatidic acid (LPA) in vitro.Our study was designed to investigate the effects of LPA on SLC26A3 expression and the diarrheal phenotype in a mouse colitis model.Methods Colitis was induced in C57BL/6 mice by adding 4% of dextran sodium sulfate (DSS) to the drinking water.The mice were assigned to LPA treatment DSS group,phosphate-buffered saline (PBS) treatment DSS group,DSS only group and untreated mice with a completely randomized design.Diarrhea severity was evaluated by measuring mice weight,disease activity index (DAI),stool water content and macroscopic evaluation of colonic damage.The effect of LPA treatment on Sic26a3 mRNA level and protein expression in the different groups of mice was investigated by quantitative PCR and Western blotting.Results All mice treated with DSS lost weight,but the onset and severity of weight loss was attenuated in the LPA treatment DSS group.The increases in stool water content and the macroscopic inflammation score in LPA treatment DSS group were significantly lower compared to DSS control group or PBS treatment DSS group ((18.89±8.67)% vs.(28.97±6.95)% or (29.48±6.71)%,P=0.049,P=0.041,respectively and 2.67±0.81 vs.4.5±0.83 or 4.5±0.54,P=0.020,P=0.006,respectively),as well as the increase in DAI (P=0.004,P=0.008,respectively).LPA enema resulted in higher Slc26a3 mRNA and protein expression levels compared to PBS-treated and untreated DSS colitis mice.Conclusion LPA increases Slc26a3 expression in the inflamed intestine and reduces diarrhea severity in DSS-induced colitis,suggesting LPA might be a therapeutic strategy in the treatment of colitis associated diarrhea.展开更多
AIM:To investigate the therapeutic effects of mesenchymal stem cells(MSCs)transplanted intraperitoneally andintravenously in a murine model of colitis.METHODS:MSCs were isolated from C57BL/6 mouse adipose tissue.MSC c...AIM:To investigate the therapeutic effects of mesenchymal stem cells(MSCs)transplanted intraperitoneally andintravenously in a murine model of colitis.METHODS:MSCs were isolated from C57BL/6 mouse adipose tissue.MSC cultures were analyzed according to morphology,cellular differentiation potential,and surface molecular markers.Experimental acute colitis was induced in C57BL/6 mice by oral administration of2%dextran sulfate sodium(DSS)in drinking water ad libitum from days 0 to 7.Colitis mice were treated with1×106 MSCs via intraperitoneal or intravenous injection on days 2 and 5.The disease activity index was determined daily based on the following parameters:weight loss,stool consistency and presence of blood in the feces and anus.To compare morphological and functional differences in tissue regeneration between different MSC injection modalities,mice were euthanized on day 8,and their colons were examined for length,weight,and histopathological changes.Inflammatory responses were determined by measuring the levels of different serum cytokines using a CBA Th1/Th2/Th17 kit.Apoptotic rates were evaluated by terminal deoxynucleotidyl transferase-mediated d UDPbiotin nick end labeling assay.RESULTS:Intravenous infusion of MSCs was more effective than intraperitoneal treatment(P<0.001)in reducing the clinical and histopathologic severity of colitis,which includes weight loss,diarrhea and inflammation.An histological evaluation demonstrated decreased colonic inflammation based on reduced crypt loss and reduced infiltration of inflammatory cells.This therapeutic effect was most likely mediated by the down-regulation of pro-inflammatory cytokines[interleukin(IL)-6 and tumor necrosis factor(TNF)];and by the up-regulation of anti-inflammatory cytokines(IL-10 and IL-4).Intravenous transplantation alsoinduced high levels of IFN that lead to activation of the immunosuppressive activity of the MSCs,which did not occur with intraperitoneal transplantation(P=0.006).An increase in apoptotic T cells was observed after intravenous,but not intraperitoneal,MSC infusion,suggesting that MSCs can induce apoptosis in resistant T cells in colonic inflammation(P=0.027).CONCLUSION:Our results demonstrate that intravenous treatment is a superior method for reducing colon inflammation compared with intraperitoneal therapy.展开更多
The purpose of this study was to investigate the possible prophylactic effects of Alpinia officinarum hance on experimentally induced acute and chronic colitis models,in-vivo and in-vitro.Acute and chronic colitis wer...The purpose of this study was to investigate the possible prophylactic effects of Alpinia officinarum hance on experimentally induced acute and chronic colitis models,in-vivo and in-vitro.Acute and chronic colitis were induced in Male Wistar rats by administration of Dextran Sulfate Sodium(DSS)in drinking water.DSS induction exhibited colon shrinkage,increased the Disease Activity Index(DAI)score,increased the levels of inflammatory markers and caused severe anemia.DSS induced animals,co-treated with the hexane extract of Alpinia officinarum(HEAO)(200 mg/kg body wt),effectively suppressed colonic injury that was evidenced by the reduced DAI score,colon weight/length ratio,histological damage,proinflammatory markers and MPO activity.Further,it restored the colonic antioxidants near to normal levels by regulating the oxidative stress via attenuation of lipid peroxidation.Our results revealed that the degree of colitis caused by the administration of DSS was significantly attenuated by HEAO.In addition,the in-vitro study showed that HEAO treatment inhibited the proliferation of HT-29 cells and down regulated the mRNA expression of NF-B and COX-2.Taken together,these results suggest that HEAO is a promising anti-oxidant and anti-inflammatory agent that support its possible therapeutic role in the treatment of colitis.展开更多
AIM To assess the therapeutic potential of Lactobacillus acidophilus(LA) for the treatment of pouchitis in a rat model.METHODS Sprague Dawley rats underwent proctocolectomy and ileal pouch-anal anastomosis followed by...AIM To assess the therapeutic potential of Lactobacillus acidophilus(LA) for the treatment of pouchitis in a rat model.METHODS Sprague Dawley rats underwent proctocolectomy and ileal pouch-anal anastomosis followed by administration of dextran sulfate sodium(DSS) to induce pouchitis. Rats with pouchitis were randomly divided into three groups: no intervention(NI), normal saline(NS, 3 m L/d normal saline for 7 d), and LA(3 m L/d LA at 1× 1010 colony-forming units for 7 d). General body condition was recorded and pouch specimens were obtained for histological examination. m RNA expression levels of interleukin(IL)-1β, IL-6, IL-10, and tumor necrosis factor-α were determined by RT-PCR. Zonula occludens protein 1(ZO-1) levels were measured by immunohistochemistry. RESULTS LA reduced weight loss associated with pouchitis(P < 0.05) and improved the symptoms of pouchitis in rats. Compared with the NI and NS groups, rats in the LAgroup showed earlier disappearance of hematochezia(6.17 ± 0.75, 6.50 ± 0.55, 3.17 ± 0.75, P < 0.05) and higher fecal scores(2.67 ± 0.48, 2.50 ± 0.51, 4.42 ± 0.50, respectively, P < 0.05). Histological scores were also lower in the LA group compared with the other two groups(7.17 ± 0.98, 8.00 ± 0.89, 4.00 ± 0.89, respectively, P < 0.05). m RNA expression levels of IL-1β, IL-6, and tumor necrosis factor-α were significantly reduced, while IL-10 m RNA levels were significantly increased in the LA group(P < 0.05, respectively). ZO-1 protein levels were also significantly increased after administration of LA(P < 0.05). CONCLUSION LA alleviates pouchitis induced by DSS after ileal pouchanal anastomosis by decreasing pro-inflammatory factors and increasing anti-inflammatory factors, and restoring ZO-1 expression in the mucosa.展开更多
BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin co...BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin complex 1(mTORC1)activation.Previous studies have demonstrated the therapeutic effects of BT2 on arthritis,liver cancer,and kidney injury.However,the effects of BT2 on ulcerative colitis(UC)are unknown.AIM To investigate the anti-UC effects of BT2 and the underlying mechanism.METHODS Mouse UC models were created through the administration of 3.5%dextran sodium sulfate(DSS)for 7 d.The mice in the treated groups were administered salazosulfapyridine(300 mg/kg)or BT2(20 mg/kg)orally from day 1 to day 7.At the end of the study,all of the mice were sacrificed,and colon tissues were removed for hematoxylin and eosin staining,immunoblot analyses,and immunohistochemical assays.Cytokine levels were measured by flow cytometry.The contents of BCAAs including valine,leucine,and isoleucine,in mouse serum were detected by liquid chromatography-tandem mass spectrometry,and the abundance of intestinal flora was analyzed by 16S ribosomal DNA sequencing.RESULTS Our results revealed that BT2 significantly ameliorated the inflammatory symptoms and pathological damage induced by DSS in mice.BT2 also reduced the production of the proinflammatory cytokines interleukin 6(IL-6),IL-9,and IL-2 and increased the anti-inflammatory cytokine IL-10 level.In addition,BT2 notably improved BCAA catabolism and suppressed mTORC1 activation and cyclooxygenase-2 expression in the colon tissues of UC mice.Furthermore,highthroughput sequencing revealed that BT2 restored the gut microbial abundance and diversity in mice with colitis.Compared with the DSS group,BT2 treatment increased the ratio of Firmicutes to Bacteroidetes and decreased the abundance of Enterobacteriaceae and Escherichia-Shigella.CONCLUSION Our results indicated that BT2 significantly ameliorated DSS-induced UC and that the latent mechanism involved the suppression of BCAA-associated mTORC1 activation and modulation of the intestinal flora.展开更多
Objective To clarify the potential mechanism of Banxia Xiexin Decoction(BXD)on colorectal cancer(CRC)from the perspective of metabolomics.Methods Forty male C57BL/6 mice were randomly divided into normal control(NC),a...Objective To clarify the potential mechanism of Banxia Xiexin Decoction(BXD)on colorectal cancer(CRC)from the perspective of metabolomics.Methods Forty male C57BL/6 mice were randomly divided into normal control(NC),azoxymethane/dextran sulfate sodium(AOM/DSS)model,low-dose BXD(L-BXD),high-dose BXD(H-BXD)and mesalamine(MS)groups according to a random number table,8 mice in each group.Colorectal cancer model was induced by AOM/DSS.BXD was administered daily at doses of 3.915(L-BXD)and 15.66 g/kg(H-BXD)by gavage for consecutive 21 days,and 100 mg/kg MS was used as positive control.Following the entire modeling cycle,colon length of mice was measured and quantity of colorectal tumors were counted.The spleen and thymus index were determined by calculating the spleen/thymus weight to body weight.Inflammatory cytokine and changes of serum metabolites were analyzed by enzyme-linked immunosorbent assay kits and ultra performance liquid chromatography-quadrupole/time-of-flight mass spectrometry(UPLC-Q/TOF-MS),respectively.Results Notably,BXD supplementation protected against weight loss,mitigated tumor formation,and diminished histologic damage in mice treated with AOM/DSS(P<0.05 or P<0.01).Moreover,BXD suppressed expression of serum inflammatory enzymes,and improved the spleen and thymus index(P<0.05).Compared with the normal group,102 kinds of differential metabolites were screened in the AOM/DSS group,including 48 potential biomarkers,involving 18 main metabolic pathways.Totally 18 potential biomarkers related to CRC were identified,and the anti-CRC mechanism of BXD was closely related to D-glutamine and D-glutamate metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,arginine biosynthesis,nitrogen metabolism and so on.Conclusion BXD exerts partial protective effects on AOM/DSS-induced CRC by reducing inflammation,protecting organism immunity ability,and regulating amino acid metabolism.展开更多
Objective To investigate the possible protective effects of sophocarpine on mucosal injury and epithelial barrier disruption on dextran sulfate sodium (DSS)-induced acute colitis. Methods Male BALB/c mice were rando...Objective To investigate the possible protective effects of sophocarpine on mucosal injury and epithelial barrier disruption on dextran sulfate sodium (DSS)-induced acute colitis. Methods Male BALB/c mice were randomly divided into three groups. The mice in normal group were given normal water, and those in model and sophocarpine- treated groups were given 2.5% DSS for 6 d to induce acute colitis. Sophocarpine (30 mg/kg) was ip administered once daily during the study period. Severity of colitis was evaluated by disease activity index (DAI), histological injury and inflammatory cytokine production including tumor necrosis factor-α (TNF-α), interleukin-lβ (IL-Iβ), and monocyte chemoattractant protein-1 (MCP-1). The colonic barrier disruption was assessed by testing the expression of junctional adhesion molecule-1 (JAM-1), E-cadherin (E-CAD), and desmocollin-2 (DSC-2) in colon mucosa. Expression of HNF4~ in colon mucosa was detected by immunohistochemistry staining and real-time RT-PCR, respectively. Results Compared with normal group, DAI, colonic shortening, and histopathological injury in model group were elevated (P 〈 0.05), but reduced in sophocarpine-treated group (P 〈 0.05). Compared with model group, the mRNA expression of inflammatory cytokines (TNF-α, IL-16, MCP-1) were obviously lower in sophocarpine-treated group (P 〈 0.05), while the cellular junction proteins (E-CAD, JAM-l, and DSC-2) were higher (P 〈 0.05). The expression of HNF4α at mRNA and protein levels was decreased significantly in model group, but increased apparently in sophocarpine-treated group. Conclusion Sophocarpine can enhance the expression of HNF4α, promote the expression of colonic intrecellular junctions, thus, maintain the integrity of the colonic barrier and inhibit the colitis process. We suggest that sophocarpine could enhance the production of cellular junction proteins to protect the intestinal barrier fuction, at least partly, in HNF4α-dependent pathway.展开更多
While inflammatory bowel disease(IBD) might be a risk factor in the development of brain dysfunctions,the underlying mechanisms are largely unknown. Here,mice were treated with 5% dextran sodium sulfate(DSS) in drinki...While inflammatory bowel disease(IBD) might be a risk factor in the development of brain dysfunctions,the underlying mechanisms are largely unknown. Here,mice were treated with 5% dextran sodium sulfate(DSS) in drinking water and sacrificed on day 7. The serum level of IL-6 increased, accompanied by elevation of the IL-6 and TNF-a levels in cortical tissue. However, the endotoxin concentration in plasma and brain of mice with DSSinduced colitis showed a rising trend, but with no significant difference. We also found significant activation of microglial cells and reduction in occludin and claudin-5 expression in the brain tissue after DSS-induced colitis.These results suggested that DSS-induced colitis increases systemic inflammation which then results in cortical inflammation via up-regulation of serum cytokines. Here,we provide new information on the impact of colitis on the outcomes of cortical inflammation.展开更多
基金supported by the Natural Science Foundation of Jiangsu Province (BK20200084)The National Natural Science Foundation of China (U1903205 and 31972971)Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province。
文摘Inflammatory bowel disease(IBD)is a complex relapsing inflammatory disease in the gut and is driven by complicated host-gut microbiome interactions.Gut commensals have shown different functions in IBD prevention and treatment.To gain a mechanistic understanding of how different commensals affect intestinal inflammation,we compared the protective effects of 6 probiotics(belonging to the genera Akkermansia,Bifidobacterium,Clostridium,and Enterococcus)on dextran sulfate sodium(DSS)-induced colitis in mice with or without gut microbiota.Anti-inflammatory properties(ratio of interleukin(IL)-10 and IL-12)of these strains were also evaluated in an in vitro mesenteric lymph nodes(MLN)co-culture system.Results showed that 4 probiotics(belonging to the species Bifidobacterium breve,Bifidobacterium bifidum,and Enterococcus faecalis)can alleviate colitis in normal mice.The probiotic strains differed in regulating the intestinal microbiota,cytokines(IL-10,IL-1βand interferon(IFN)-γ),and tight junction function(Zonulin-1 and Occludin).By constrast,Akkermansia muciniphila AH39 and Clostridium butyricum FHuNHHMY49T1 were not protective.Interestingly,B.breve JSNJJNM2 with high anti-inflammatory potential in the MLN model could relieve colitis symptoms in antibiotic cocktail(Abx)-treated mice.Meanwhile,E.faecalis FJSWX25M1induced low levels of cytokines in vitro and showed no beneficial effects.Therefore,we provided insight into the clinical application of probiotics in IBD treatment.
基金Supported by National Natural Science Foundation of China,No.31370921Natural Science Foundation of Liaoning Province,No.2015020515
文摘AIM To assess the effect of sodium selenite on the severity of dextran sulfate sodium(DSS)-induced colitis in C57BL/6 mice.METHODS Mice were randomly divided into four groups(n = 10/group): normal group, selenium(Se) group, chronic colitis group, and Se + chronic colitis group. The mice were sacrificed on day 26. Survival rates, clinical symptoms, colon length, and histological changes were determined. The percentages and absolute numbers of immune system cells in the lamina propria lymphocytes(LPL) of the colon, the expression of m RNA in colon tissue, and the concentrations of Th1, Th17, and Treg cytokines in LPL from the large intestine, were measured.RESULTS Se significantly ameliorated the symptoms of colitis and histological injury(P < 0.05 each), increasing the proportions of neutrophils and CD4+ CD25+ T cells(P < 0.05 each) and decreasing the proportions of γδT cells, CD4+, CD4+CD44+, and CD4+ CD69+ T cells in LPL(P < 0.05 each). Moreover, Se reduced the expression of IL-6, IFN-γ, IL-17 A, IL-21, T-bet, and RORγt(P < 0.05 each), but enhanced the expression of IL-10 and Foxp3(P < 0.05 each). CONCLUSION These results suggest that Se protects against DSSinduced chronic colitis perhaps by increasing the number of CD4(+)CD25(+) Tregs that suppress the secretion of proinflammatory cytokines and populations of Th1, Th17, and γδT cells.
基金Supported by the 13th Five-Year Plan for National Key R&D Program of China,No. 2018YFC1705405Scientific Research Innovation Team Project of Beijing University of Chinese Medicine,No. 2019-JYB-TD004+1 种基金New Faculty Startup Fund Program of BUCM,No. 2022-JYB-XJSJJ078National Natural Science Foundation of China,No. 82004113
文摘BACKGROUND Intestinal micro-ecological imbalances impair the intestinal barrier and induce intestinal inflammation,for example,ulcerative colitis(UC).According to the latest research,abnormalities in intestinal microbiota structure and their metabolites play a dominant role in UC progression;in addition,they could affect the mucus barrier based on different factors.Although numerous studies have confirmed the important role of intestinal microbiota in UC pathogenesis,the intricate connection between microbiota and metabolites and mucus barrier in UC occurrence remains unclear,and correlation analyses of differential microbiota and their metabolites under UC are relatively scarce.AIM To reveal the differential intestinal microbiota and metabolites in UC pathogenesis and explore more sensitive biomarker compositions.METHODS We used the antibiotic combination method to establish intestinal pseudo-aseptic mice;afterward,dextran sulfate sodium(DSS)was applied to establish an acute experimental colitis mice model.Colitis severity,assessed based on disease activity index,colorectal length,colorectal wet weight,and histological lesions,and mucus-related staining(mucopolysaccharide alcian blue and immunofluorescence of mucin),was compared between the pseudo-aseptic and bacterial colitis mice.Finally,differential intestinal microbiota,metabolites,and their association and correlations,were analyzed by 16s rDNA sequencing in combination with non-targeted metabolomics,through gas chromatography-mass spectrometry.RESULTS Compared with the pseudo-aseptic mice,intestinal bacteria positive mice were more severely ill and their intestinal mucus loss was more pronounced in DSS-induced colitis(P<0.05),suggesting that different microbiota and metabolites could cause the different degrees of colitis.Subsequently,we observed that in addition to Klebsiella,and Bacteroides,which were widely associated with colitis,Candidatus Stoquefichus,Anaerobiospirillum,Muribaculum,and Negativibacillus may be involved in protection against colitis.Furthermore,differential metabolites of the microbiota were mainly enriched in the synthesis-related pathways of key structural sequences of mucin.In combination with the mucin-related staining and immunofluorescence results,the findings indicate that the differential microbiota and their metabolites potentially regulate the composition and function of mucus under colitis.CONCLUSION Microbiota and their metabolites are major factors regulating the composition and function of mucus,in turn influencing the function and structure of intestinal mucus barrier under colitis.The different microbiota and metabolites identified in the present study could be novel biomarkers for colitis.
基金the grants from the National Natural Science Foundation of China(30800812)
文摘Ulcerative colitis (UC) is a lifelong illness with profound emotional and social impacts, and could cause serious damage to large intestine, especially in colon. However, the pathogenesis of UC remained unclear. The present study attempts to find out the role of matrix metalloproteinases-7 (MMP-7) and lysozyme in the pathogenesis of UC through a mice model induced by dextran sulfate sodium (DSS). The UC model was evaluated both by disease activity index (DAI) and the intestinal histopathology. The results show that there is a high correlation between the DAI score and the pathological changes of colon. Interleukin-6 (IL-6) serum levels and large intestinal fluids levels in UC mice are always higher than that of the control groups, which might be associated with the degree of the inflammation damage in the colon. The change tendency of the MMP-7 mRNA and protein expressions are both up-regulated firstly and then down-regulated from 1 to 5 d in the colon, but only the MMP-7 protein is up-regulated at 7 d again. The up-regulated MMP-7 levels in the early stage of UC may play a protective role through the activated defensins, while the down-regulated levels in the mid-later stage of UC may be connected with the severe lesions in the colon. However, the up-regulated MMP-7 levels in the later stage of UC in the colon may also contribute to the tissue repair or be served as a marker to CRC (colorectal cancer). The distribution of lysozyme protein indicates that there may be Paneth-like cells in the colon. Both the changes of MMP-7 and lysozyme in the small intestine may play a protective role for the safe environment of the whole gut, especially to the colon of UC.
文摘Background: Disrupted Ca2+ homeostasis contributes to the development of colonic dysmotility in ulcerative colitis (UC), but the underlying mechanisms are unknown. This study aimed to examine the alteration of colonic smooth muscle (SM) Ca2+ signaling and Ca2+ handling proteins in a rat model of dextran sulfate sodium (DSS)-induced UC. Methods: Male Sprague-Dawley rats were randomly divided into control (n = 18) and DSS (n = 17) groups. Acute colitis was induced by 5% DSS in the drinking water for 7 days. Contractility of colonic SM strips (controls, n = 8 and DSS, n = 7) was measured in an organ bath. Cytosolic resting Ca2+ levels (n = 3 in each group) and Ca2+ transients (n = 3 in each group) were measured in single colonic SM cells. Ca2+ handling protein expression was determined by Western blotting (n = 4 in each group). Differences between control and DSS groups were analyzed by a two-sample independent t-test. Results: Average tension and amplitude of spontaneous contractions of colonic muscle strips were significantly enhanced in DSS-treated rats compared with controls (1.25 ± 0.08 g vs. 0.96 - 0.05 g, P = 0.007; and 2.67 - 0.62 g vs. 0.52 ±0.10 g, P= 0.013). Average tensions of carbachol-evoked contractions were much weaker in the DSS group (1.08 ±0.10 g vs. 1.80 ±0.19 g, P = 0.006). Spontaneous Ca2+ transients were observed in more SM cells from DSS-treated rats (15/30 cells) than from controls (5/36 cells). Peak caffeine-induced intracellular Ca2+ release was lower in SM cells of DSS-treated rats than controls (0.413 ±0.046 vs. 0.548 ±0.041, P = 0.033). Finally, several Ca2+ handling proteins in colonic SM were altered by DSS treatment, including sarcoplasmic reticulum calcium-transporting ATPase 2a downregulation and phospholamban and inositol 1,4,5-trisphosphate receptor 1 upregulation. Conclusions: Impaired intracellular Ca2+ signaling of colonic SM, caused by alteration of Ca2+ handing proteins, contribute to colonic dysmotility in DSS-induced UC.
基金Traditional Korean Medicine R&D Program funded by the Ministry of Health&Welfare through the Korea Health Industry Development Institute(KHIDI),No.HI15C00255and National Research Foundation of Korea(NRF)funded by the Korean government(MSIP),No.2018R1A5A2025272.
文摘BACKGROUND Sulfasalazine has been used as a standard-of-care in ulcerative colitis for decades,however,it results in severe adverse symptoms,such as hepatotoxicity,blood disorders,male infertility,and hypospermia.Accordingly,the new treatment strategy has to enhance pharmacological efficacy and stimultaneously minimize side effects.AIM To compare the anti-inflammatory action of sulfasalazine alone or in combination with herbal medicine for ulcerative colitis in a dextran sodium sulfate(DSS)-induced colitis mouse model.METHODS To induce ulcerative colitis,mice received 5%DSS in drinking water for 7 d.Animals were divided into five groups(n=9 each)for use as normal(non-DSS),DSS controls,DSS+sulfasalazine(30 mg/kg)-treatment experimentals,DSS+sulfasalazine(60 mg/kg)-treatment experimentals,DSS+sulfasalazine(30 mg/kg)+Citrus unshiu peel and Bupleuri radix mixture(30 mg/kg)(SCPB)-treatment experimentals.RESULTS The SCPB treatment showed an outstanding effectiveness in counteracting the ulcerative colitis,as evidenced by reduction in body weight,improvement in crypt morphology,increase in antioxidant defenses,down-regulation of proinflammatory proteins and cytokines,and inhibition of proteins related to apoptosis.CONCLUSIONSCPB may represent a promising alternative therapeutic against ulcerative colitis,without inducing adverse effects.
基金Supported by Guangdong Administration of Traditional Chinese Medicine(No.201 01 92)
文摘Objective:To investigate the effect of total alkaloids of Sophora alopecuroides(TASA) on dextran sulfate sodium(DSS)-induced colitis in mice.Methods:Chronic experimental colitis was induced by administration of 4 cycles of 4%DSS.Fifty mice were randomly distributed into 4 groups(normal,DSS,DSS/high-dose TASA, and DSS/low-dose TASA groups) by a random number table with body weight stratification.Mice in the normal group(n=11) and DSS-induced colitis control group(n=15) received control treatment of 20 mL/kg distilled water; DSS plus TASA high- and low-dose groups(n=12 each) were treated with TASA solution(20 mL/kg) at the doses of 60 mg/kg and 30 mg/kg,respectively.The severity of colitis was assessed on the basis of clinical signs, colon length,and histology scores.Moreover,secretory immunoglobulin A(slgA) and haptoglobin(HP) were analyzed by enzyme linked immunosorbent assay;intercellular adhesion molecule 1(ICAM-1) and macrophage-migration inhibitory factor(MIF) gene expressions were analyzed by quantitative reverse transcriptase realtime polymerase chain reaction(qRT-PCR) using SYBA greenⅠ;and nuclear factorκB(NF-κB) expression and activation and p65 interaction with the promoter of ICAM-1 gene were assessed by Western blotting and chromatin immunoprecipitation assay.Results:TASA administration significantly attenuated the damage and substantially reduced HP elevation and maintained the level of cecum slgA.TASA inhibited the ICAM-1 gene expression and had no effect on MIF gene expression.Also,TASA was able to reduce phospho-lκBα(p-lκBα) protein expression;however,it had no effect on the activation of IκB kinaseα(IKKα) and inhibitor of NF-κBα(IκBα).Moreover,TASA inhibited the p65 recruitment to the ICAM-1 gene promoter.Conclusions:TASA had a protective effect on DSS-induced colitis.Such effect may be associated with its inhibition of NF-κB activation and blockade of NF-κB-regulated transcription activation of proinflammatory mediator gene.
基金supported by a grant from the Natural Science Foundation of Liaoning province, China (No. 20072101)
文摘The role of interleukin 25 (IL-25) in a number of human diseases still has not been extensively studied, here we attempt to evaluate the role of recombinant IL-25 (rIL-25) in the development of dextran sulfate sodium (DSS)- induced experimental colitis. Acute colitis was induced in female C57BL/6 mice by oral administration of 2.5% DSS in drinking water ad libitum. At the same time as the start of DSS exposure, mice were injected intraperitoneally with 0.4 +tg of rIL-25 or PBS. Then disease activity index (DAI), histological changes and survival rate were observed. The levels of IL-17, IL-23, and TGF-β1 in colon tissues were determined by ELISA, and the production of IL-17 by CD4+/CD8+ T cells was detected by intracellular flow cytometry. In contrast to the DSS treated mice, DSS + rIL-25 treated mice displayed a lower DAI, limited histological changes and prolonged survival. The levels of IL-23 and TGF-β1 were significantly elevated in the DSS + rIL-25 treated mice compared to the DSS treated mice. There was no significant difference in the production of IL-17 in colon tissues and CD4+/CD8+ T cells between the DSS + rIL-25 treated mice and DSS treated mice. Our findings suggest the role of IL-25 in inhibiting development and progression of acute colitis in DSS-induced mouse colitis model. Cellular & Molecular Immunology. 2008;5(6):425-431.
基金This work was supported by a grant from the Natural Science Foundation of Zhejiang Province (No. Y2110864). Conflicts of interest: None.
文摘Background It has been reported that CD8+ regulatory cells could be induced upon oral tolerance. The purpose of this study was to investigate the changes of CD8a+ T cells in dextran sulfate sodium (DSS)-induced colitis mice pretreated by oral immune regulation. Methods The effects of five low oral doses of colitis-extracted proteins (CEP) on colitis were evaluated by clinical manifestation and histological lesions. The percentages of CD8a+ T cells gating on CD3+ T cells were evaluated in the gut-associated lymphoid tissues (GALT) and the spleens by flow cytometry. Differences between the two groups were compared by Student's t test or Mann-Whitney U test. Results Compared to bovine serum albumin (BSA)-fed control mice, administration of CEP resulted in marked alleviation of colitis. The proportion of CDSa+ T ceils, not only in intraepithelial lymphocytes (IELs) and lamina propria lymphocytes (LPLs) of the large intestine (LI) but also in spleen from CEP-fed colitis mice, was significantly higher than that from BSA-fed colitis mice (LI-IELs: (71.5±5.4)% vs. (60.1±4.3)%, P 〈0.01; LI-LPLs: (60.7±5.2)% vs. (51.9±4.7)%, P 〈0.01; spleen: (24.1±3.6)% vs. (20.3±4.1)%, P 〈0.05; n=8). Mucosal repair in repair-period mice five days after termination of DSS treatment was also accompanied by an increase of CD8a+ T cells in large intestinal mucosal lymphocytes (LI-IELs: (72.1±3.7)% vs. (61.5±4.5)%, P 〈0.01; LI-LPLs: (62.1±5.7)% vs. (52.7±3.6)%, P 〈0.01; n=8). The proportion of CD3+ T cells increased in Peyer's patches (PPs) and decreased in mesenteric lymph nodes (MLNs) from colitis mice compared to untreated mice, whereas the change pattern of CD3+T cells in PPs and MLNs from CEP-fed colitis mice was just on the contrary. Conclusion Improvement of DSS-induced colitis resulted from oral immune regulation is associated with an increase in CD8a+T cells in spleen and large intestinal mucosa.
基金Supported by the Department of Veterans Affairs,Office of Research and Development(Biomedical Laboratory Research and Development)No.BX001155
文摘Inflammatory bowel diseases(IBD),including Crohn's disease and ulcerative colitis,are complex diseases that result from the chronic dysregulated immune response in the gastrointestinal tract. The exact etiology is not fully understood,but it is accepted that it occurs when an inappropriate aggressive inflammatory respon-se in a genetically susceptible host due to inciting environmental factors occurs. To investigate the path-ogenesis and etiology of human IBD,various animal models of IBD have been developed that provided indispensable insights into the histopathological and morphological changes as well as factors associated with the pathogenesis of IBD and evaluation of therapeutic options in the last few decades. The most widely used experimental model employs dextran sodium sulfate(DSS) to induce epithelial damage. The DSS colitis model in IBD research has advantages over other various chemically induced experimental models due to its rapidity,simplicity,reproducibility and controllability. In this manuscript,we review the newer publicized advances of research in murine colitis models that focus upon the disruption of the barrier function of the intestine,effects of mucin on the development of colitis,alterations found in microbial balance and resultant changes in the metabolome specifically in the DSS colitis murine model and its relation to the pathogenesis of IBD.
基金supported by grants from the National Natural Science Foundation Youth Project of China(No.81703716)Jiangxi Science Foundation for Distinguished Young Scholars(No.20224ACB216019)+5 种基金the Natural Science Foundation of Jiangxi Province(No.20202BABL206151 and No.20202BABL216026)Youth Talents Project of Jiangxi Science and Technology Normal University(No.2017QNBJRC006)Doctoral Startup Fund of Jiangxi Science and Technology Normal University(No.2019BSQD015)Department Education Science and Technology Research Project of Jiangxi(No.GJJ201134)the Open Project of Jiangxi Provincial Key Laboratory of Drug Design and Evaluation(No.JKD-KF-2104)the National Undergraduate Training Program for Innovation of China(No.202211318024).
文摘Objective Patients with chronic ulcerative colitis(UC)often have mental symptoms such as depression and anxiety,and stress can lead to gastrointestinal diseases.However,the correlation between mental stress and UC is unclear.In this paper,chronic unpredictable mild stress(CUMS)was utilized to evaluate the involvement of mental factors in the pathogenesis of UC.Methods The CUMS model was used to evaluate the direct/indirect involvement of mental factors in the pathogenesis of UC.The behavior was evaluated by the open field,forced swimming,and tail suspension tests.Body weight,the disease activity index(DAI)score,colon length,and HE staining of colon tissue were used to evaluate the action of CUMS and fluoxetine.Results The results showed that weight loss and the DAI score increased in CUMS mice,but they had no meaningful effect on colon length and morphological structure of colon tissue.However,CUMS aggravated dextran sulfate sodium(DSS)-induced colon length shortening and colon morphological structure damage.Fluoxetine significantly improved the DAI score,shortened colon length,and damaged morphology and structure of the colons induced by CUMS combined with DSS in mice.Fluoxetine also decreased the level of IL-6 in the serum and the TNF-αand IFN-γlevels of colon tissue.Fluoxetine simultaneously improved behavioral abnormalities induced by CUMS combined with DSS in mice.Conclusion CUMS aggravated the UC symptoms induced by DSS,and fluoxetine could improve the UC symptoms due to its improvement in the inflammatory level and behavioral abnormalities.
基金This study was supported by grants from National Natural Science Foundation of China (No. 81100264, No. 81360076, and No. 81100498), the Hubei Provincial Health Department Foundation for Young Scholars (QJX2012-04) and the Deutsche Forschungsgemeinschaft SFB621/C9 (to US).Acknowledgment: The authors thank the participants involved in this study and Dr. Ma Ruling for helping with data analysis.
文摘Background Diarrhea is a common clinical feature of ulcerative colitis resulting from unbalanced intestinal fluid and salt absorption and secretion.The Cl-/HCO3-exchanger SLC26A3 is strongly expressed in the mid-distal colon and plays an essential role in colonic Cl-absorption and HCO3-secretion.Sic26a3 expression is up-regulated by lysophosphatidic acid (LPA) in vitro.Our study was designed to investigate the effects of LPA on SLC26A3 expression and the diarrheal phenotype in a mouse colitis model.Methods Colitis was induced in C57BL/6 mice by adding 4% of dextran sodium sulfate (DSS) to the drinking water.The mice were assigned to LPA treatment DSS group,phosphate-buffered saline (PBS) treatment DSS group,DSS only group and untreated mice with a completely randomized design.Diarrhea severity was evaluated by measuring mice weight,disease activity index (DAI),stool water content and macroscopic evaluation of colonic damage.The effect of LPA treatment on Sic26a3 mRNA level and protein expression in the different groups of mice was investigated by quantitative PCR and Western blotting.Results All mice treated with DSS lost weight,but the onset and severity of weight loss was attenuated in the LPA treatment DSS group.The increases in stool water content and the macroscopic inflammation score in LPA treatment DSS group were significantly lower compared to DSS control group or PBS treatment DSS group ((18.89±8.67)% vs.(28.97±6.95)% or (29.48±6.71)%,P=0.049,P=0.041,respectively and 2.67±0.81 vs.4.5±0.83 or 4.5±0.54,P=0.020,P=0.006,respectively),as well as the increase in DAI (P=0.004,P=0.008,respectively).LPA enema resulted in higher Slc26a3 mRNA and protein expression levels compared to PBS-treated and untreated DSS colitis mice.Conclusion LPA increases Slc26a3 expression in the inflamed intestine and reduces diarrhea severity in DSS-induced colitis,suggesting LPA might be a therapeutic strategy in the treatment of colitis associated diarrhea.
基金Supported by Fundo de IncentivoàPesquisa e Eventos do Hospital de Clínicas de Porto Alegre and Coordenacao de Aperfeicoamento de Pessoal de Nível Superior
文摘AIM:To investigate the therapeutic effects of mesenchymal stem cells(MSCs)transplanted intraperitoneally andintravenously in a murine model of colitis.METHODS:MSCs were isolated from C57BL/6 mouse adipose tissue.MSC cultures were analyzed according to morphology,cellular differentiation potential,and surface molecular markers.Experimental acute colitis was induced in C57BL/6 mice by oral administration of2%dextran sulfate sodium(DSS)in drinking water ad libitum from days 0 to 7.Colitis mice were treated with1×106 MSCs via intraperitoneal or intravenous injection on days 2 and 5.The disease activity index was determined daily based on the following parameters:weight loss,stool consistency and presence of blood in the feces and anus.To compare morphological and functional differences in tissue regeneration between different MSC injection modalities,mice were euthanized on day 8,and their colons were examined for length,weight,and histopathological changes.Inflammatory responses were determined by measuring the levels of different serum cytokines using a CBA Th1/Th2/Th17 kit.Apoptotic rates were evaluated by terminal deoxynucleotidyl transferase-mediated d UDPbiotin nick end labeling assay.RESULTS:Intravenous infusion of MSCs was more effective than intraperitoneal treatment(P<0.001)in reducing the clinical and histopathologic severity of colitis,which includes weight loss,diarrhea and inflammation.An histological evaluation demonstrated decreased colonic inflammation based on reduced crypt loss and reduced infiltration of inflammatory cells.This therapeutic effect was most likely mediated by the down-regulation of pro-inflammatory cytokines[interleukin(IL)-6 and tumor necrosis factor(TNF)];and by the up-regulation of anti-inflammatory cytokines(IL-10 and IL-4).Intravenous transplantation alsoinduced high levels of IFN that lead to activation of the immunosuppressive activity of the MSCs,which did not occur with intraperitoneal transplantation(P=0.006).An increase in apoptotic T cells was observed after intravenous,but not intraperitoneal,MSC infusion,suggesting that MSCs can induce apoptosis in resistant T cells in colonic inflammation(P=0.027).CONCLUSION:Our results demonstrate that intravenous treatment is a superior method for reducing colon inflammation compared with intraperitoneal therapy.
文摘The purpose of this study was to investigate the possible prophylactic effects of Alpinia officinarum hance on experimentally induced acute and chronic colitis models,in-vivo and in-vitro.Acute and chronic colitis were induced in Male Wistar rats by administration of Dextran Sulfate Sodium(DSS)in drinking water.DSS induction exhibited colon shrinkage,increased the Disease Activity Index(DAI)score,increased the levels of inflammatory markers and caused severe anemia.DSS induced animals,co-treated with the hexane extract of Alpinia officinarum(HEAO)(200 mg/kg body wt),effectively suppressed colonic injury that was evidenced by the reduced DAI score,colon weight/length ratio,histological damage,proinflammatory markers and MPO activity.Further,it restored the colonic antioxidants near to normal levels by regulating the oxidative stress via attenuation of lipid peroxidation.Our results revealed that the degree of colitis caused by the administration of DSS was significantly attenuated by HEAO.In addition,the in-vitro study showed that HEAO treatment inhibited the proliferation of HT-29 cells and down regulated the mRNA expression of NF-B and COX-2.Taken together,these results suggest that HEAO is a promising anti-oxidant and anti-inflammatory agent that support its possible therapeutic role in the treatment of colitis.
基金Supported by Jie-Shou Li Gut Barrier Foundation,No.LJS_201008
文摘AIM To assess the therapeutic potential of Lactobacillus acidophilus(LA) for the treatment of pouchitis in a rat model.METHODS Sprague Dawley rats underwent proctocolectomy and ileal pouch-anal anastomosis followed by administration of dextran sulfate sodium(DSS) to induce pouchitis. Rats with pouchitis were randomly divided into three groups: no intervention(NI), normal saline(NS, 3 m L/d normal saline for 7 d), and LA(3 m L/d LA at 1× 1010 colony-forming units for 7 d). General body condition was recorded and pouch specimens were obtained for histological examination. m RNA expression levels of interleukin(IL)-1β, IL-6, IL-10, and tumor necrosis factor-α were determined by RT-PCR. Zonula occludens protein 1(ZO-1) levels were measured by immunohistochemistry. RESULTS LA reduced weight loss associated with pouchitis(P < 0.05) and improved the symptoms of pouchitis in rats. Compared with the NI and NS groups, rats in the LAgroup showed earlier disappearance of hematochezia(6.17 ± 0.75, 6.50 ± 0.55, 3.17 ± 0.75, P < 0.05) and higher fecal scores(2.67 ± 0.48, 2.50 ± 0.51, 4.42 ± 0.50, respectively, P < 0.05). Histological scores were also lower in the LA group compared with the other two groups(7.17 ± 0.98, 8.00 ± 0.89, 4.00 ± 0.89, respectively, P < 0.05). m RNA expression levels of IL-1β, IL-6, and tumor necrosis factor-α were significantly reduced, while IL-10 m RNA levels were significantly increased in the LA group(P < 0.05, respectively). ZO-1 protein levels were also significantly increased after administration of LA(P < 0.05). CONCLUSION LA alleviates pouchitis induced by DSS after ileal pouchanal anastomosis by decreasing pro-inflammatory factors and increasing anti-inflammatory factors, and restoring ZO-1 expression in the mucosa.
基金Supported by National Natural Science Foundation of ChinaNo. 82074241+1 种基金Project of Jiangsu Province Hospital of Traditional Chinese Medicine Peak TalentNo. y2021rc36
文摘BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin complex 1(mTORC1)activation.Previous studies have demonstrated the therapeutic effects of BT2 on arthritis,liver cancer,and kidney injury.However,the effects of BT2 on ulcerative colitis(UC)are unknown.AIM To investigate the anti-UC effects of BT2 and the underlying mechanism.METHODS Mouse UC models were created through the administration of 3.5%dextran sodium sulfate(DSS)for 7 d.The mice in the treated groups were administered salazosulfapyridine(300 mg/kg)or BT2(20 mg/kg)orally from day 1 to day 7.At the end of the study,all of the mice were sacrificed,and colon tissues were removed for hematoxylin and eosin staining,immunoblot analyses,and immunohistochemical assays.Cytokine levels were measured by flow cytometry.The contents of BCAAs including valine,leucine,and isoleucine,in mouse serum were detected by liquid chromatography-tandem mass spectrometry,and the abundance of intestinal flora was analyzed by 16S ribosomal DNA sequencing.RESULTS Our results revealed that BT2 significantly ameliorated the inflammatory symptoms and pathological damage induced by DSS in mice.BT2 also reduced the production of the proinflammatory cytokines interleukin 6(IL-6),IL-9,and IL-2 and increased the anti-inflammatory cytokine IL-10 level.In addition,BT2 notably improved BCAA catabolism and suppressed mTORC1 activation and cyclooxygenase-2 expression in the colon tissues of UC mice.Furthermore,highthroughput sequencing revealed that BT2 restored the gut microbial abundance and diversity in mice with colitis.Compared with the DSS group,BT2 treatment increased the ratio of Firmicutes to Bacteroidetes and decreased the abundance of Enterobacteriaceae and Escherichia-Shigella.CONCLUSION Our results indicated that BT2 significantly ameliorated DSS-induced UC and that the latent mechanism involved the suppression of BCAA-associated mTORC1 activation and modulation of the intestinal flora.
基金Supported by the Natural Science Foundation of Nanjing University of Chinese Medicine(No.XZR2020038)Science and Technology Innovation Project of Suzhou Medical and Health Care(No.SKJY2021136)Fifth Batch of Gusu Health Personnel Training Project in Suzhou(No.GSWS2020085)。
文摘Objective To clarify the potential mechanism of Banxia Xiexin Decoction(BXD)on colorectal cancer(CRC)from the perspective of metabolomics.Methods Forty male C57BL/6 mice were randomly divided into normal control(NC),azoxymethane/dextran sulfate sodium(AOM/DSS)model,low-dose BXD(L-BXD),high-dose BXD(H-BXD)and mesalamine(MS)groups according to a random number table,8 mice in each group.Colorectal cancer model was induced by AOM/DSS.BXD was administered daily at doses of 3.915(L-BXD)and 15.66 g/kg(H-BXD)by gavage for consecutive 21 days,and 100 mg/kg MS was used as positive control.Following the entire modeling cycle,colon length of mice was measured and quantity of colorectal tumors were counted.The spleen and thymus index were determined by calculating the spleen/thymus weight to body weight.Inflammatory cytokine and changes of serum metabolites were analyzed by enzyme-linked immunosorbent assay kits and ultra performance liquid chromatography-quadrupole/time-of-flight mass spectrometry(UPLC-Q/TOF-MS),respectively.Results Notably,BXD supplementation protected against weight loss,mitigated tumor formation,and diminished histologic damage in mice treated with AOM/DSS(P<0.05 or P<0.01).Moreover,BXD suppressed expression of serum inflammatory enzymes,and improved the spleen and thymus index(P<0.05).Compared with the normal group,102 kinds of differential metabolites were screened in the AOM/DSS group,including 48 potential biomarkers,involving 18 main metabolic pathways.Totally 18 potential biomarkers related to CRC were identified,and the anti-CRC mechanism of BXD was closely related to D-glutamine and D-glutamate metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,arginine biosynthesis,nitrogen metabolism and so on.Conclusion BXD exerts partial protective effects on AOM/DSS-induced CRC by reducing inflammation,protecting organism immunity ability,and regulating amino acid metabolism.
文摘Objective To investigate the possible protective effects of sophocarpine on mucosal injury and epithelial barrier disruption on dextran sulfate sodium (DSS)-induced acute colitis. Methods Male BALB/c mice were randomly divided into three groups. The mice in normal group were given normal water, and those in model and sophocarpine- treated groups were given 2.5% DSS for 6 d to induce acute colitis. Sophocarpine (30 mg/kg) was ip administered once daily during the study period. Severity of colitis was evaluated by disease activity index (DAI), histological injury and inflammatory cytokine production including tumor necrosis factor-α (TNF-α), interleukin-lβ (IL-Iβ), and monocyte chemoattractant protein-1 (MCP-1). The colonic barrier disruption was assessed by testing the expression of junctional adhesion molecule-1 (JAM-1), E-cadherin (E-CAD), and desmocollin-2 (DSC-2) in colon mucosa. Expression of HNF4~ in colon mucosa was detected by immunohistochemistry staining and real-time RT-PCR, respectively. Results Compared with normal group, DAI, colonic shortening, and histopathological injury in model group were elevated (P 〈 0.05), but reduced in sophocarpine-treated group (P 〈 0.05). Compared with model group, the mRNA expression of inflammatory cytokines (TNF-α, IL-16, MCP-1) were obviously lower in sophocarpine-treated group (P 〈 0.05), while the cellular junction proteins (E-CAD, JAM-l, and DSC-2) were higher (P 〈 0.05). The expression of HNF4α at mRNA and protein levels was decreased significantly in model group, but increased apparently in sophocarpine-treated group. Conclusion Sophocarpine can enhance the expression of HNF4α, promote the expression of colonic intrecellular junctions, thus, maintain the integrity of the colonic barrier and inhibit the colitis process. We suggest that sophocarpine could enhance the production of cellular junction proteins to protect the intestinal barrier fuction, at least partly, in HNF4α-dependent pathway.
基金supported by grants from the National Natural Science Foundation of China (81430044)the National Basic Research Development Program of China (2012CB518200 and 2011CB910800)
文摘While inflammatory bowel disease(IBD) might be a risk factor in the development of brain dysfunctions,the underlying mechanisms are largely unknown. Here,mice were treated with 5% dextran sodium sulfate(DSS) in drinking water and sacrificed on day 7. The serum level of IL-6 increased, accompanied by elevation of the IL-6 and TNF-a levels in cortical tissue. However, the endotoxin concentration in plasma and brain of mice with DSSinduced colitis showed a rising trend, but with no significant difference. We also found significant activation of microglial cells and reduction in occludin and claudin-5 expression in the brain tissue after DSS-induced colitis.These results suggested that DSS-induced colitis increases systemic inflammation which then results in cortical inflammation via up-regulation of serum cytokines. Here,we provide new information on the impact of colitis on the outcomes of cortical inflammation.