Immune Modulatory Treatment of Trinitrobenzene Sulfonic Acid Colitis with Calcitriol Is Associated with a Change of a T Helper (Th) 1/Th17 to a Th2 and Regulatory T Cell Profile (original) (raw)
Related papers
Trinitrobenzene sulfonic acid (TNBS)-induced colitis is one of the most common methods for studying inflammatory bowel disease in animal models. Several factors may, however, affect its reproducibility, rate of animal mortality, and macroscopic and histopathological outcomes. Our aim was to validate the main contributing factors to this method and compare the effects of different reference drugs upon remission of resultant colon injuries. TNBS was dissolved in 0.25 ml of ethanol (50% v/v) and instilled (25, 50, 100 and 150 mg/kg) intracolonically to the male Wistar rats. After determination of optimum dose of TNBS in male rats and assessment of this dose in female rats, they were treated with reference drugs including dexamethasone [1 mg/kg, intraperitoneally (i.p.) and 2 mg/kg, orally (p.o.)], Asacol (mesalazine, 100 mg/kg, p.o.; 150 mg/kg, enema) and hydrocortisone acetate (20 mg/kg, i.p.; 20 mg/kg, enema) which started 2 h after colitis induction and continued daily for 6 consecutive days. Thereafter, macroscopic and microscopic parameters and clinical features were assessed and compared in different groups. We found that the optimum dose of TNBS for the reproducibility of colonic damage with the least mortality rate was 50 mg/kg. Amongst studied reference drugs, hydrocortisone acetate (i.p.), dexamethasone (i.p. and p.o.) and Asacol (p.o.) significantly diminished the severity of macroscopic and microscopic injuries and could be considered effective for experimental colitis studies in rats. Our findings suggest that optimization of TNBS dose is essential for induction of colitis under the laboratory conditions; and gender exerts no impact upon macroscopic and histological characteristics of TNBS-induced colitis in rats. Furthermore, the enema forms of hydrocortisone and Asacol are not appropriate reference drugs.
Biological & Pharmaceutical Bulletin, 2011
Ulcerative colitis (UC) and Crohn's disease (CD), major inflammatory bowel diseases (IBD) affecting the gastrointestinal tract in humans, are characterized by chronic and spontaneously relapsing inflammation. While the etiology of IBD is not fully understood, early histological and functional changes as well as late permanent tissue destruction are known to be induced by mucosal injury in the colon caused by inflammatory responses generated by persistent and inadequate mucosal innate and adoptive immune reactions against intestinal bacteria, as well as by environmental factors in genetically susceptible individuals. 1,2) While systemic glucocorticoids (GCs) are considered the current first-line therapy for patients with severe UC, approximately 30% of patients do not respond to GC therapy. 3) In recent years, oral treatment with tacrolimus has been proven effective in inducing remission in patients with refractory UC, thereby allowing for the reduction or withdrawal of GC. 4,5) However, while tacrolimus is known to inhibit production of interleukin (IL)-2, IL-4, IL-5, and interferon (IFN)-g in activated peripheral T cells through inhibition of calcineurin activity, 6) little is known regarding its effect on the production of cytokines in inflamed colon tissue. It is shown that in rodents, acute colitis was induced during dextran sulfate sodium (DSS) treatment, mainly through activating innate immune cells such as neutrophils and macrophages, 7) and colitis was perpetuated via the adaptive immune response, primarily by T cell activation, even after cessation of DSS treatment. 8) Melger et al. showed that colon tissue in the chronic phase of DSS-induced colitis shared some pathophysiological aspects with human UC, such as ulceration, edema, dense infiltration of mononuclear cells, irregular epithelial formation with crypt distortion, and goblet cell depletion. 9) Given these findings, the DSS-induced colitis model is believed to be suitable for evaluating the therapeutic potential of drugs for treating UC. Here, using inflamed colons of colitic mice, we investigated the effect of tacrolimus on extant colitis and on the release of IFN-g, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-12, and tumor necrosis factor (TNF)-a, cytokines pivotal in inducing and perpetuating colitis in DSS-treated mice and humans with UC. 9) MATERIALS AND METHODS Drugs Solid dispersion formulation of tacrolimus and its placebo were prepared by Astellas Pharma Inc. (Tokyo, Japan). These compounds were suspended in distilled water and diluted to appropriate concentrations at the time of use. Animals Six-week-old male C57BL/6J mice, purchased from Charles River Laboratories Japan, Inc. (Kanagawa, Japan), were maintained in a temperature-and humidity-controlled room with a 12-h light-dark cycle. All animal experimental procedures were approved by the Institutional Animal Care and Use Committee of Astellas Pharma Inc. and were in accordance with regulations laid down by the Association for the Assessment and Accreditation of Laboratory Animal Care. Induction of Colitis DSS (molecular weight: 36000 to 50000; MP Biomedicals, Solon, OH, U.S.A.) was dissolved in distilled water at a concentration of 2.5% (w/v). Colitis was induced in mice by administering the DSS solution as drinking water for seven days (the first day of DSS treatment and the day of treatment completed were designated as Day 0 and Day 7, respectively); distilled water was given to mice in the normal group for the same period. On Day 10, 3 d after cessation of DSS treatment, mice showing loss of body weight were selected for the investigation. All mice were then divided into groups in order to obtain similar mean rates of body weight loss for each group. Colitic mice numbered 8 to 10 per group, while normal mice numbered 3 to 5 of normal mice in a group were included in each study. Time-Course Study of Colitis Four sets of normal (nϭ4) and DSS-treated control groups (nϭ8) were prepared
Clinical & Experimental Immunology, 2011
Tumour necrosis factor (TNF)-a plays a critical role in the pathogenesis of T helper type 1-mediated colitis such as Crohn's disease. However, the roles of its two receptors in mediating pathology remain largely unknown. In this study, trinitrobenzene sulphonic acid (TNBS) was used to induce colitis in TNF-receptor single or double knock-out (DKO) BALB/c mice and in wildtype counterparts. TNF-R1 -/mice had significantly less weight loss, reduced mortality, colon shortening and oedema, colon histological damage and lower levels of colon myeloperoxidase compared with wild-type (WT) BALB/c mice. A similar manifestation was also observed in TNF-R2 -/and TNF-R1 -/-TNF-R2 -/-(TNF-R DKO) mice. Strikingly, systemic inflammatory response (including splenomegaly and monocyte expansion) was found in WT and TNF-R1 -/mice after TNBS, instead of TNF-R2 -/and TNF-R DKO mice.
2014
The 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced acute inflammatory bowel disease (IBD) model in the rat is discussed, focusing on the details of the TNBS instillation and highlighting the advantages and limitations of this model. For determination of the time- dependent action of 50% ethanol and different doses of TNBS, male Wistar rats were treated with 50% ethanol or 10 mg or 30 mg of TNBS dissolved in 50% ethanol. The TNBS-induced inflammation peaked 48-72 h after installation and the colitis caused by 30 mg of TNBS was more severe than that caused by 10 mg of TNBS. To test the effectiveness of sulfasalazine (SASP), male rats were treated with 10 mg of TNBS or with 10 mg of TNBS and SASP, and 72 h later the extent of mucosal damage was determined. Orally administered 50 mg/kg/day SASP proved to reduce the TNBS-induced colonic inflammation in rats significantly. The TNBS-induced colitis model facilitates a better understanding of the immunopathological mechanisms of IBD. Op-...
Protective effects of citicoline on TNBS-induced experimental colitis in rats
International journal of clinical and experimental medicine, 2014
The aim of this study was to investigate the effects of citicoline on the development of colitis and antioxidant parameters in rats subjected to tribenzene sulfonic acid (TNBS)-induced colitis. Twenty four Wistar Albino female rats were divided into four subgroups (n=6) (control, colitis control, colitis + 50 mg/kg citicoline, colitis + 250 mg/kg citicoline). Colitis was induced using an enema of TNBS and ethanol; following which citicoline was administrated for 3 days and effects of citicoline was subsequently evaluated. Based on microscopic damage scores, there was no difference between rats of the TNBS-colitis and 50 mg/kg citicoline treated groups, whereas treatment with 250 mg/kg citicoline, caused significant reduction in colon injury compared to that observed in rats of TNBS-colitis group. In terms of the biochemical analyses, myeloperoxidase (MPO), malondialdehyde (MDA), reduced glutathione (GSH), and IL-6 levels in rats from 250 mg/kg citicoline group were significantly dif...
Digestive Diseases and Sciences, 2014
Recent findings indicate that carbon monoxide (CO) in non-toxic doses exerts a beneficial antiinflammatory action in various experimental models. However, the precise anti-inflammatory mechanism of CO in the intestine remains unclear. Here, we assessed the effects of a novel water-soluble CO-releasing molecule, CORM-3, on trinitrobenzene sulfonic acid (TNBS)induced colitis in mice. Methods To induce colitis, C57BL/6 male mice received an enema of TNBS. CORM-3 or its inactive compound, iCORM-3, were administered intraperitoneally, once immediately before, and twice daily after receiving an enema of TNBS. Three days after TNBS administration, the distal colon was removed, assessed for colonic damage and histological scores, polymorphonuclear leukocyte recruitment (tissue-associated myeloperoxidase, MPO activity), and TNF-a, IFN-c and IL-17A expression (mRNA and protein levels in the colon mucosa). CD4 ? T cells isolated from murine spleens were stimulated with anti-CD3/CD28, in the presence or absence of CORM-3/ iCORM-3. The cell supernatants were assessed for TNF-a and IFN-c expression, 24 h following stimulation. Results Colonic damage and histological scores were significantly increased in TNBS-induced mice compared to sham-operated mice. Tissue-associated MPO activity and expression of TNF-a, IFN-c, and IL-17A in the colonic mucosa were higher in TNBS-induced colitis mice. The above changes were attenuated in CORM-3-treated mice. Further, CORM-3 was effective in reducing TNF-a and IFN-c production in anti-CD3/CD28-stimulated CD4 ? T cells.
Immunology, 2016
The clinical benefits of short-term therapy with glucocorticoids (GC) in Inflammatory Bowel Disease (IBD) patients are widely known. However, the effects of this treatment towards the reestablishment of the regulatory network in IBD are not fully explored yet. Therefore, here we evaluated the immunological effects of the abbreviated GC therapy in experimental colitis induced by 3% Dextran Sulfate Sodium (DSS) in C57BL/6 mice. Treatment with GC improved disease outcome, constrained circulating leukocytes and ameliorated intestinal inflammation. The control of the local inflammatory responses involved a reduction in the expression of IFN-γ and IL-1ß, associated to augmented mRNA levels of peroxisome proliferator-activated receptors PPAR (α and γ) in intestine. Furthermore, there was a reduction of CD4(+) T cells producing IFN-γ, together with an increased frequency of the putative regulatory population of T cells producing IL-10, in spleen. These systemic alterations were accompanied ...
Immunomodulatory Action of a Proprietary Product Using TNBS (Tri Nitro Benzene Sulfonic Acid) - Induced Ulcerative Colitis in Sprague Dawley Rats, 2021
Immunomodulatory activity of the novel formulation with respect to gut health using TNBS induced ulcerative colitis was studied in the present experiment. The action of Biofield Energy Healing (the Trivedi Effect®) based test formulation and Biofield Energy Treatment per se was studies. The test formulation consisted of essential ingredients viz. minerals (zinc, magnesium, iron, and copper) and vitamins (B6, B12, and D3). Each ingredient of the test formulation was divided into two parts. One part was denoted as the control without any Biofield Energy Healing Treatment, while the other part was defined as the Biofield Energy Treated sample, which received the Biofield Energy Healing Treatment by a renowned Biofield Energy Healer, Mr. Mahendra Kumar Trivedi. Additionally, three group of animals also received Biofield Energy Treatment per se (day -15) under similar conditions. The immune biomarkers such as immunoglobulin M (IgM), IgG, IgA, IgE, biochemistry, and hematology parameters were evaluated. The level of IgE was significantly increased by 22.95%, 25.51%, and 48.57% (p≤0.001) in the G5, G6, and G7 groups, respectively as compared with the untreated test group (G4). IgM level was significantly (p≤0.001) increased by 12.16%, 20.27%, and 11.49% in the G7, G8, and G9 groups, respectively as compared with the G4 group. The level of IgG was significantly (p≤0.001) increased by 11.19% in the G9 group as compared with the G4 group. Hematology data suggested significant increased the levels of total leukocyte count (TLC) (25.95%; p≤0.05 in G9), neutrophils (65.26%, 59.62%, and 117.84% (p≤0.05) in G6, G7, and G9 respectively), and monocytes (61.97% in G9) were reported when compared with the G4 group. Further, the level of lymphocytes were significantly (p≤0.001) increased by 98.24%, 70.31%, 61.91%, and 108.20% in the G5, G6, G8, and G9 groups, respectively as compared to the disease control (G2) group. Serum glutamic-oxaloacetic transaminase (SGOT) or AST level was significantly reduced by 31.96% (p≤0.05), 23.24%, 23.44%, 24.45%, and 23.93% in the G5, G6, G7, G8, and G9 groups, respectively as compared with the G2 group. Moreover, serum glutamic pyruvic transaminase (SGPT) or ALT level was reduced by 22.63% and 23.44% in G6 group as compared to G2 and G6 groups, respectively. The level of alkaline phosphatase (ALP) was reduced by 28.42%, 21.50%, 17.91%, 17.00%, and 21.63% in the G5, G6, G7, G8, and G9 groups, respectively than G2 group. Creatine kinase-myocardial band (CK-MB) level was significantly (p≤0.001) decreased by 46.26%, 28.88%, 42.33%, 43.57%, and 43.97%, respectively in G5, G6, G7, G8, and G9 groups, respectively as compared with the G4 group. The level of testosterone was increased by 45.96% and 35.31% in the G6 group as compared with the G2 and G4 groups, respectively. In addition, animal body weight, feed intake, and relative organ weight data did not showed any abnormal findings with respect to the safe and non-toxic treatment strategies. In conclusion, the Trivedi Effect®-Biofield Energy Healing Treatment and the test formulation has the significant capacity for immunomodulatory effect, which supposed to be found effective against overall gut health and inflammation.
Journal of Interferon & Cytokine Research, 2010
IBD is characterized by uncontrolled immune responses in inflamed mucosa, with dominance of IL-17-producing cells and deficiency of Treg cells. The aim of this study was to explore the effect and mechanisms of RA, the ligand of RAR␣, on immune responses in human and murine colitis. Colonic biopsies from patients with UC were cultured and treated with RA as the agonist of RAR␣ or LE135 as the antagonist of RAR␣. Expressions of IL-17 and FOXP3 were detected by immunohistochemistry. Murine colitis was induced by intrarectal administration with TNBS at Day 1. Mice were then i.p.-treated with RA or LE135 daily for 7 days. Cytokine levels in the cultures of mouse LPMCs were measured. Expressions of FOXP3 and IL-17 in colon tissues or MLN were detected by immunohistological analysis. Body weight and colon inflammation were evaluated. RA treatment up-regulated FOXP3 expression and downregulated IL-17 expression in colon biopsies of patients and in colon tissues and MLN of mice with colitis compared with controls. LPMCs from RA-treated mice produced lower levels of proinflammatory cytokines (TNF-␣, IL-1, IL-17) but more regulatory cytokines (IL-10, TGF-) compared with that of untreated mice. LE135 showed the opposite effect of RA. Furthermore, RA ameliorated TNBS-induced colitis in a dose-dependent manner, as seen by improved body weight and colon inflammation. RA down-regulates colon inflammatory responses in patients with IBD in vitro and in murine colitis in vivo, representing a potential therapeutic approach in IBD treatment. J. Leukoc. Biol. 86: 959 -969; 2009. Article 0741-5400/09/0086-959