Colonic Wet Weight Research Articles

Effects of polyphenolic maqui (Aristotelia chilensis) extract on the inhibition of NLRP3 inflammasome and activation of mast cells in a mouse model of Crohn's disease-like colitis.

Crohn's disease (CD) involves activation of mast cells (MC) and NF-кB in parallel with the PPAR-α/NLRP3 inflammasome/IL-1β pathway in the inflamed colon. Whether polyphenols from maqui (Aristotelia chilensis) represent a natural alternative treatment for CD is unclear. Therefore, we used an animal model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced CD-like colitis to investigate protective effects of maqui extract through monitoring NLRP3 inflammasome and MC activation in colon tissue. Maqui extract was administered via orogastric route to mice after (post-Treatment group) or prior (pre-Treatment group) to TNBS-induction. Colon pathology was characterized by histoarchitectural imaging, disease activity index (DAI), and assessing NF-кB, p-NF-кB, PPAR-α/NLRP3 expression and IL-1β levels. Compared to mice treated with TNBS alone administration of anthocyanin-rich maqui extract improved the DAI, colon histoarchitecture and reduced both colon wet-weight and transmural inflammation. Induction with TNBS significantly increased colonic NLPR3 inflammasome activation, while co-treatment with maqui extract (either post- or pre-Treatment) significantly downregulated NLRP3, ASC and caspase-1 levels, which manifested as reduced colonic IL-1β levels. Supplemented maqui extract marginally diminished NF-кB activity in epithelial cells but reached statistical significance in immune cells (as judged by decreased NF-кB phosphorylation). PPAR-α signaling was largely unaffected by Maqui whereas MC infiltration into the colon mucosa and submucosa decreased and their level of degranulation was suppressed. These outcomes show the post- and pre- Treatment effect of a polyphenolic extract rich in anthocyanins from maqui the acute phase of TNBS- induced CD-like colitis is linked to suppression of the NLRP3 inflammasome and reduced MC responses. These data indicate that maqui extract represents a potential nutraceutical for the treatment of inflammatory bowel disease (IBD).

Effects of Prebiotic Fructooligosaccharides on Susceptibility to Dextran Sodium Sulfate‐Induced Colitis

Inflammatory bowel disease (IBD) is a chronic intestinal disease that includes Crohn’s disease and ulcerative colitis. Previous studies have found alterations in the microbiome in IBD and in animal models of IBD, underscoring the role of the intestinal microbiota in IBD pathogenesis. Prebiotics are molecules in foods, such as fructooligosaccharides and other soluble fiber molecules, that support the growth or activity of beneficial probiotic microbes. Previous studies suggest that prebiotics can reduce symptoms in models of colitis. This study aimed to determine whether pretreatment with 1% short‐chain fructooligosacchardies (FOS) for 1 week, followed by 8 days of 3% DSS administration with continued FOS treatment, altered the severity of disease in C57Bl/6 mice. Clinical signs of colitis, including weight loss, fecal occult blood, and fecal consistency, were monitored daily during the DSS treatment period and the daily disease activity index was calculated. At the end of the experiment, samples were collected for histological scoring, and for RNA isolation and analysis of colonic pro‐inflammatory cytokine expression by RT‐PCR. Through the 8 days of DSS treatment, the disease activity index increased in both DSS‐treated and FOS/DSS‐treated mice compared to control non‐colitic mice, but was not significantly different between the two colitic groups. In addition, colon wet weight, colon length, and histological colitis scores were similar in colitic mice treated with FOS compared to colitic mice without FOS treatment. Colonic claudin mRNA expression was lower in DSS‐treated mice than control mice, but was not significantly altered by FOS treatment. These data suggest that treatment with fructooligosaccharides did not significantly alter susceptibility to DSS‐induced colitis.

Alhagi pseudalhagi Extract Exerts Protective Effects Against Intestinal Inflammation in Ulcerative Colitis by Affecting TLR4-Dependent NF-κB Signaling Pathways.

Alhagi pseudalhagi Desv. Extract (APE) is the major active fraction extracted from the aerial part of Alhagi pseudalhagi Desv. In view of its application in Uyghur medicine, it may be beneficial for the treatment of ulcerative colitis (UC). The aim of the present study was to investigate the possible beneficial effects of APE on UC mice and detect the possible mechanisms underlying these effects. Methods: An acute UC model was established in mice using dextran sulfate sodium. Sixty mice were randomly divided into six groups: normal, UC model, sulfasalazine (200 mg/kg), high-dose APE (APE-H, 2.82 g/kg), middle-dose APE (APE-M, 1.41 g/kg), and low-dose APE (APE-L, 0.70 g/kg) groups. Drugs were administered by gavage for 10 days after the induction of colitis. Serum and colon tissue samples were collected from the mice during the experiment, and survival signs, body weight changes, disease activity index (DAI), colon length, and colon wet weight in mice were determined after the treatment. UC-induced damage, including inflammation and ulceration of colon mucosa, were observed by the naked eye as well as using hematoxylin and eosin staining (H&E) and scanning electron microscopy and scored according to Wallace and Keean’s criteria. We measured the levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6, and IL-10 in the serum and colon tissues using ELISA. Additionally, the relative protein levels of toll-like receptor 4 (TLR4), nuclear factor-kappa B p65 (NF-κB p65), phosphorylated NF-κB p65 at Ser536 (p-p65 Ser536), inhibitor kappa B-kinase ß (IK-Kβ), and phosphorylated IK-Kβ (Ser176/180) (p-IK-Kβ) in colonic mucosal epithelial tissues were detected using western blotting. The main functional components of APE were analyzed and confirmed by UPLC-MS/MS. Results: APE treatment repaired the UC-induced colon mucosa injury, reduced the weight loss, attenuated DAI, colon macroscopic damage index, and histological inflammation, and significantly downregulated the levels of inflammatory markers, including TNF-α, IL-1β, and IL-6, in the serum and colon tissues. Additionally, APE treatment reduced the levels of TLR4 and phosphorylation of p-NF-κB and p-IK-Kβ. The main components of APE are taxifolin, 3,5-dihydroxy-2-(4-hydroxyphenyl)-7-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl) oxan-2-yl] oxychromen-4-one, hyperoside, rutin, kaempferol, isorhamnetin, 7,8-dihydroxyflavone, and kaempferide. Conclusions: To the best of our knowledge, the present study is first to demonstrate that APE exerts a protective effect against intestinal inflammation in UC by affecting TLR4-dependent NF-κB signaling pathways.

Gabapentin improves colonic inflammatory damage and oxidative stress on acetic acid-induced colitis independent of cannabinoid pathway

Objective: Gabapentin (GBP) possess a systemic anti-inflammatory action confirmed and endocannabinoids system have been effectives in reduces inflammatory disorders on intestinal tract. The aim of this study is evaluate the participation of the endogenous cannabinoids pathway in the anti-inflammatory and antioxidant effects of GBP in acetic acid (AA) induced colitis in mice. Methods: Colitis induction was performed using AA (6%) and mice treated intraperitoneally with GBP or dexamethasone (subcutaneously) at 17 h or 17:30 h after induction of colitis, respectively. After 18 h of colitis induction, the animals were euthanized and colonic sample was taken for evaluation of macroscopic and microscopic lesion, wet weight and biochemical analyzes. Results: The administration of GBP was effective in reduce the macro and microscopic lesions, significant decreasing the colonic wet weight of colitis mice. The drug reduced the concentration myeloperoxidase (MPO), malondialdehyde (MDA) and interleukin 1 (IL-1) and increased the level glutathione (GSH) in colitis mice compared to normal mice. The treatment with endocannabinoids (ECs) receptors antagonists did not alter the GBP effect. Conclusion: Gabapentin was able to reduce inflammatory parameters in acid acetic-induced colitis, but its effect seems to be independent of cannabinoid pathway.

Effects of Combined Inulin and VSL#3 on Susceptibility to Dextran Sodium Sulfate‐Induced Colitis

Inflammatory bowel disease (IBD) is a chronic intestinal disease that includes Crohn's disease and ulcerative colitis. Previous studies have found alterations in the microbiome in IBD and in animal models of IBD, underscoring the role of the intestinal microbiota in IBD pathogenesis. Prebiotics are molecules in foods that support the growth or activity of beneficial microbes. Inulin, a soluble fiber molecule, is a commonly available prebiotic, and previous studies suggest that it can reduce symptoms in dextran sodium sulfate (DSS)-induced colitis in mice. VSL#3 is a commercially available mixture of 8 probiotic species that has shown beneficial effects in models of intestinal inflammation. This study aimed to determine whether pretreatment with 1% inulin for 1 week, followed by 7 days of 3% DSS administration with inulin plus VSL#3 treatment, altered the severity of disease in C57Bl/6 mice. Clinical signs of colitis, including weight loss, fecal occult blood, and fecal consistency, were monitored daily during the DSS treatment period. At the end of the experiment, samples were collected for histological scoring, and for RNA isolation and analysis of colonic pro-inflammatory cytokine expression by RT-PCR. At the end of DSS treatment, the disease activity index was not different between control colitic mice or those treated with inulin and VSL#3. In addition, colon length and colon wet weight were similar in the two groups. Treatment with inulin+VSL#3 did not significantly alter expression of mRNA for COX-2, TGF-beta, or KC in the colon of mice with DSS colitis. In conclusion, a combination of inulin + VSL#3 treatment did not significantly alter susceptibility to DSS-induced colitis.

McN-A-343, a muscarinic agonist, reduces inflammation and oxidative stress in an experimental model of ulcerative colitis

AimThe aim of the present study was to investigate the anti-inflammatory response mediated of the M1 muscarinic acetylcholine receptor (mAChR) during experimental colitis. Material and methodsAfter the induction of 6% acetic acid colitis, mice were treated with McN-A-343 0.5, 1.0, and 1.5 mg/kg or dexamethasone (DEXA, 2.0 mg/kg) or pirenzepine (PIR, 10 mg/kg; M1 mAChR antagonist). Colonic inflammation was assessed by macroscopic and microscopic lesion scores, colonic wet weight, myeloperoxidase (MPO) activity, interleukin-1 beta (IL1-β) levels and tumor necrosis factor alpha (TNF-α), glutathione (GSH), malondialdehyde (MDA) and nitrate and nitrite (NO3/NO2), mRNA expression of IKKα, nuclear factor kappa beta (NF-kB) and cyclooxygenase-2 (COX-2), as well protein expression of NF-kB and COX-2. ResultsTreatment with McN-A-343 at a concentration of 1.5 mg/kg showed a significant reduction in intestinal damage as well as a decrease in wet weight, MPO activity, pro-inflammatory cytokine concentration, markers of oxidative stress and expression of inflammatory mediators. The action of the M1 agonist by the administration of pirenzepine, which promoted the blocking of the mAChR M1-mediated anti-inflammatory response, has also been proven. ConclusionThe results suggest that peripheral colonic M1 mAChR is involved in reversing the pro-inflammatory effect of experimentally induced colitis, which may represent a promising therapeutic alternative for patients with ulcerative colitis.

Blumea lacera DC., accelerates the healing of acetic acid induced ulcerative colitis in rats by regulating oxidative stress and colonic inflammation: in-vivo and in silico molecular docking experiments

To expound the protective outcome of standardised hydroalcoholic extract of Blumea lacera (EEBL) in acetic acid-induced ulcerative colitis (UC). Further, in silico computational experiments were performed to identify the inhibitory activity of bioactive constituents of EEBL against inflammatory targets. Rats were pre-treated with EEBL (100 and 200 mg/kg) for 7 days and on eighth day, UC was induced by intrarectal instillation of 2 ml of 4% v/v acetic acid and treatment continues till 11th day. Colonic injury was examined by clinical activity score and mucosal inflammation was assessed by colon wet weight, macroscopic scoring and histopathological examination. In addition, colonic contractility studies and biochemical analysis were carried out. Further, bioactive compounds from title plant were subjected to drug likeness prediction and molecular docking studies against myeloid differentiation factor-2 (MD-2), nuclear factor-kappa B (NF-κB), tumor necrosis factor alpha (TNF-alpha) and soluble epoxide hydrolase (sEH). Pre-treatment with both the doses of EEBL significantly ameliorated the acetic acid-induced colonic injury and these results were parallels with biochemical and histopathology examinations. Out of fifteen selected bioactive compounds, 5-hydroxy-3methyl-3,6,7,4-tetramethoxy flavones, lachnophyllic acid and d-fenchone exhibited better drug likeness property with higher docking score. The observed protective outcome might be recognized due to the synergistic effect of identified bioactive compounds by inhibition of key targets of inflammation.

Effects of prebiotic inulin pre‐treatment on susceptibility to DSS‐induced colitis in mice

Inflammatory bowel disease (IBD) is a chronic intestinal disease that includes Crohn’s disease and ulcerative colitis. Previous studies have found alterations in the microbiome in IBD and in animal models of IBD, underscoring the role of the intestinal microbiota in IBD pathogenesis. Prebiotics are molecules in foods that support the growth or activity of beneficial microbes. Inulin, a soluble fiber molecule, is a commonly available prebiotic, and previous studies suggest that it can reduce symptoms in dextran sodium sulfate (DSS)‐induced colitis in mice. This study aimed to determine whether pretreatment with 1% inulin for 1 week, followed by 7 days of 3% DSS administration with continued inulin treatment, altered the severity of disease in C57Bl/6 mice. Clinical signs of colitis, including weight loss, fecal occult blood, and fecal consistency, were monitored daily during the DSS treatment period. At the end of the experiment, samples were collected for histological scoring, and for RNA isolation and analysis of colonic pro‐inflammatory cytokine expression by quantitative PCR. Unexpectedly, inulin pre‐treated, DSS‐exposed mice were positive for blood in the stool a day sooner than mice given DSS only. However, by the end of DSS treatment, the disease activity index was not different between the two groups. In addition, colon length and colon wet weight were similar in inulin‐treated mice and non‐inulin treated mice. Histological scoring of colitis showed no difference between inulin‐treated and non‐inulin‐treated colitic mice. In conclusion, 7 days of pre‐treatment with inulin followed by induction of acute DSS‐induced colitis did not significantly alter susceptibility to colitis.Support or Funding InformationSupported by the Missouri Western State University PORTAL program

6]-Gingerol modulates spermatotoxicity associated with ulcerative colitis and benzo[a]pyrene exposure in BALB/c mice.

The deterioration of male reproductive health may represent an outcome of an active disease and environmental factors. The present study investigated the modulatory role of [6]-gingerol in spermatotoxicity resulting from colitis and benzo[a]pyrene (B[a]P), an environmental and food-borne pollutant. Group I (control) mice received corn oil alone, while group II ([6]-gingerol alone) mice orally received [6]-gingerol alone at 100 mg/kg body weight. Group III [benzo[a]pyrene+dextran sulfate sodium (BDS) alone] mice were orally exposed to B[a]P at 125 mg/kg for 7 days followed by three cycles of 4% dextran sulfate sodium (DSS) in drinking water. A cycle consisted of seven consecutive days of exposure to DSS-treated water followed by 14 consecutive days of normal drinking water. Group IV (BDS+[6]-gingerol) mice were orally treated daily with 100 mg/kg of [6]-gingerol during exposure to B[a]P and DSS in the same manner as those of group III. [6]-Gingerol significantly abrogated BDS-mediated increase in disease activity index and restored the colon wet weight, colon length and colon mass index to near normal when compared to BDS alone group. Moreover, [6]-gingerol significantly prevented BDS-induced decreases in the daily sperm production (DSP), testicular sperm number (TSN), epididymal sperm number, sperm progressive motility and sperm membrane integrity when compared with the control. [6]-Gingerol markedly increased the sperm antioxidant enzymes activities and decreased the sperm head, mid-piece and tail abnormalities as well as suppressed oxidative stress and inflammatory biomarkers in BDS-exposed mice. [6]-Gingerol protected against spermatotoxicity in experimental model of interaction of colitis with environmental pollutant B[a]P.

Effect of Niacin on Inflammation and Angiogenesis in a Murine Model of Ulcerative Colitis

Butyrate and niacin are produced by gut microbiota, however butyrate has received most attention for its effects on colonic health. The present study aimed at exploring the effect of niacin on experimental colitis as well as throwing some light on the ability of niacin to modulate angiogenesis which plays a crucial role of in the pathogenesis of inflammatory bowel disease. Rats were given niacin for 2 weeks. On day 8, colitis was induced by intrarectal administration of iodoacetamide. Rats were sacrificed on day 15 and colonic damage was assessed macroscopically and histologically. Colonic myeloperoxidase (MPO), tumour necrosis factor (TNF)-α, interleukin (IL)-10, vascular endothelial growth factor (VEGF), angiostatin and endostatin levels were determined. Niacin attenuated the severity of colitis as demonstrated by a decrease in weight loss, colonic wet weight and MPO activity. Iodoacetamide-induced rise in the colonic levels of TNF-α, VEGF, angiostatin and endostatin was reversed by niacin. Moreover, niacin normalized IL-10 level in colon. Mepenzolate bromide, a GPR109A receptor blocker, abolished the beneficial effects of niacin on body weight, colon wet weight as well as colonic levels of MPO and VEGF. Therefore, niacin was effective against iodoacetamide-induced colitis through ameliorating pathologic angiogenesis and inflammatory changes in a GPR109A-dependent manner.

Tangshen Formula Attenuates Colonic Structure Remodeling in Type 2 Diabetic Rats.

Aim. This study investigated the effect and mechanism of the Chinese herbal medicine Tangshen Formula (TSF) on GI structure remodeling in the rat model of diabetes. Methods. Type 2 diabetic rats were used. Wet weight per unit length, layer thicknesses, levels of collagens I and III, nuclear factor kappa B (NF-κB), interferon-γ (IFN-γ), interleukin-6 (IL-6), transforming growth factor-β1 (TGF-β1), and Smad2/3 expression in the rat colon were measured. Results. Compared with the control group animals, wet weight and layer thicknesses of the colon increased, and expressions of collagens I and III, NF-κB, IFN-γ, IL-6, TGF-β1, and Smad2/3 increased significantly in the diabetic animals. TSF inhibited increase in colonic wet weight and layer thicknesses, downregulated expressions of collagens I and III in the mucosal layer, and downregulated expressions of NF-κB, IFN-γ, IL-6, TGF-β1, and Smad2/3 in the colon wall. Furthermore, level of expression of NF-κB was associated with those of TGF-β1 and Smad2/3. Expression of TGF-β1 was associated with the most histomorphometric parameters including colonic weight, mucosal and muscle thicknesses, and levels of collagens I and III in mucosal layer. Conclusion. TSF appears to attenuate colonic structure remodeling in type 2 diabetic rats through inhibiting the overactivated pathway of NF-κB, thus reducing expressions of TGF-β1.

Therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis

Objective To investigate the therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis. Methods A total of 40 Sprague-Dawley rats were divided into four groups including the healthy control group, model group, fecal transplant treatment group and vancomycin treatment group. Rats in three experimental groups were subcutaneously injected with clindamycin phosphate (10 mg), followed by treatment with toxin producing Clostridium difficile (ACTT43255) enema 24 hours later. The rats in fecal transplant treatment group and vancomycin treatment group were respectively treated with fecal suspension and vancomycin one day after modeling. The rats were fasted for one day after the last administration and then executed. The levels of potassium ion (K), sodium ion (Na), albumin (ALB), white blood cells (WBC), C-reaction protein (CRP), interleukin-1β (IL-1β), interleukin-10 (IL-10), interleukin-12 (IL-12) and interleukin-17 (IL-17) as well as the percentage of neutrophils (N%) in serum samples were detected. The colon tissue samples were collected for pathology examination. Results The rat model of pseudomembranous colitis was successfully established by subcutaneous injection of clindamycin in combination with toxin-producing Clostridium difficile (ACTT43255) enema. The signs of intestinal inflammation including serious weight loss, remarkably shortened colon length and significantly increased colon wet weight index were observed in rats from the model group (P<0.05). Compared with the rats from model group, the rats received fecal transplant showed significantly increased levels of K, ALB, IL-10 and IL-10/IL-12 in serum and decreased levels of WBC, N%, CRP, IL-1β and IL-17 (P<0.05). Conclusion Fecal transplantation was proved to be an effective approach for the treatment of pseudomembranous colitis. The therapeutic mechanism might due to its impacts on serum inflammatory factors. Key words: Fecal transplantation; Pseudomembranous colitis; Inflammatory factor

Active components alignment of Gegenqinlian decoction protects ulcerative colitis by attenuating inflammatory and oxidative stress

Ethnopharmacological relevanceGegenqinlian Decoction (GQD) has been used as a folk remedy for gastrointestinal diseases in China over thousands of years. It has significant treatment efficacy for patients with inflammatory bowel disease (IBD). We analyzed and showed that the active components alignment of Gegenqinlian Decoction (ACAG) possesses broad pharmacological effects including analgesic, antipyretic, anti-inflammatory, antibacterial, antiviral and antidiarrhea, as well as the effect of adjusting gastrointestinal function in our preliminary experiments. However, the exact molecular mechanisms on how ACAG exerts these pharmacological effects still remain elusive. In the present study, the plausible pharmacological effects of ACAG on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis were investigated. Materials and methodsMale Sprague-Dawley (SD) rats with TNBS/ethanol-induced colitis were used. The colonic wet weight, macroscopic and histological colon injury, superoxide dismutase (SOD), malonyldialdehyde (MDA), and inducible nitric oxide synthase (iNOS) activity were observed. Pro-inflammation cytokines were determined by ELISA methods, semi-quantitative RT-PCR and Immuno-histochemistry. ResultsWe showed administration of ACAG was able to improve colitis. This was manifested by a decreased in the score of macroscopic and histological colonic injury, by lowered colonic wet weight, accompanied by significant increased of SOD activity, and decreased of MDA and iNOS activities. The treatment also significantly reduced tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) levels in colon and serum as well as the colonic mRNA levels for several inflammatory cytokines such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), macrophage inflammatory protein-2 (MIP-2), intercellular adhesion molecule-1 (ICAM-1) and toll-like receptor 2, 4 (TLR2, TLR4). In addition, we also showed that ACAG was able to inhibit the activation and translocation of transcription factors, nuclear factor kappaBp65 (NF-κBp65) in colon. ConclusionsOur results suggest that ACAG exhibits protective effect in TNBS-induced ulcerative colitis. We postulate that this might be due to its modulation of oxidant/anti-oxidant balance, downregulation of productions, expressions of pro-inflammatory cytokines and inhibition of NF-κBp65 signal transduction pathways.

Amentoflavone inhibits iNOS, COX-2 expression and modulates cytokine profile, NF-κB signal transduction pathways in rats with ulcerative colitis

Ulcerative colitis is a chronic inflammatory disorder characterized by oxidative stress, leucocyte infiltration and upregulation of pro-inflammatory cytokines. The aim of the present study was to examine the effect of amentoflavone on a murine model of ulcerative colitis (UC). UC was induced by intracolonic injection of 3% acetic acid in male Wistar rats. amentoflavone (10 mg/kg·b.wt) or reference drug sulfasalazine (100 mg/kg·b.wt) was administrated intra-peritoneally for 5 consecutive days before induction of colitis with acetic acid. Administration of amentoflavone was found to reduce the extent of inflammatory colonic injury. This was manifested by a decrease in the score of mucosal injury, by lowered colonic wet weight as well as vascular permeability and diminished lactate dehydrogenase (LDH) and myeloperoxidase (MPO) activity reflecting reduced leukocyte infiltration. Furthermore, the mucosal content of lipid peroxidation (LPO), glutathione (GSH), superoxide dismutase (SOD), nitric oxide (NO) activity confirms that amentoflavone could significantly inhibit colitis. The treatment also reduced significantly the colonic tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and IL-6 levels as well as the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) compared to colitis control group. The histopathological studies also confirm the foregoing findings. amentoflavone was also able to inhibit the activation and translocation of transcription factors, nuclear factor (NF)-κB subunits (p65/p50). These results suggest that amentoflavone exhibits protective effect in acetic acid-induced ulcerative colitis which might be due to its modulation of oxidant/anti-oxidant balance, down-regulation of productions and expressions of pro-inflammatory cytokines, inflammatory mediators and inhibition of NF-κB signal transduction pathways.

The possible mechanisms of Picrasma quassiodes (D. Don) Benn. in the treatment of colitis induced by 2,4,6-trinitrobenzene sulfonic acid in mice

Ethnopharmacological relevancePicrasma quassiodes (D. Don) Benn.(PQB) is used in folk medicines for the treatment of colds, upper respiratory infection, acute tonsillitis, acute gastroenteritis, bacillary dysentery and a variety of acute infectious diseases in Asia. Although recent reports indicate that PQB has antibacterial, and anti-inflammatory effects, its effects on colitis and its inhibitory mechanisms have not been previously reported. Aim of the studyTo assess the effects and the mode of action of the extract of Picrasma quassiodes (D. Don) Benn.(PQB) on a model of colitis in mice induced by trinitrobenzene sulfonic acid (TNBS). Materials and methodsWe induced mice colitis using TNBS/ethanol, then different doses of Picrasma quassiodes (D. Don) Benn.(PQB) extract (100, 200 and 400mg/kg/day) and sulfasalazine (500mg/kg/day) were administered by gavage for 7 days after the induction of colitis. The mice body weight, colonic wet weight, colonic lengths, myeloperoxidase (MPO) activity, macroscopic and histological colon injury were observed. Pro-inflammatory cytokines such as: tumor necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8) were assayed by enzyme-linked immunoassay. The protein expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in the colons were determined by immunohistochemical analysis. ResultsPQB administration effectively prevented mice diarrhea, decreasing of the body weights, shortening of colon length and increasing of colon wet weight. Macroscopic and histological examinations also indicated that it was protected against colonic edema, ulceration and MPO activity elevation. Furthermore, PQB inhibited the abnormal secretions of pro-inflammatory cytokines, such as TNF-α and IL-8. Additionally, administration of PQB effectively inhibited COX-2 and iNOS protein expression. ConclusionsThese results suggest that PQB has an anti-inflammatory effect on TNBS-induced colitis due to the down-regulations of the productions and expressions of inflammatory mediators, and that it may be a potential inflammatory bowel disease (IBD) drug candidate.

The protective and healing effects of Si Shen Wan in trinitrobenzene sulphonic acid-induced colitis

Ethnopharmacological relevanceSi Shen Wan is a traditional Chinese herbal medicine formula for the treatment of diseases with diarrhea, such as ulcerative colitis, allergic colitis and chronic colitis. To investigate the protective and healing effects of Si Shen Wan in the experimental colitis induced by trinitrobenzene sulphonic acid, and to furture explore its mechanism of action. Materials and methodsRats with colitis treated with Si Shen Wan for 10 days. Colon wet weight, colon organ coefficient, colonic damage score and pathological change after trinitrobenzene sulphonic acid challenge were determined. The levels of MPO, MDA, GSH-PX, SOD and the expression of IL-4 and IL-10 mRNA in the colon were also measured. ResultsAfter treatment, colon wet weight, colon organ coefficient and colonic damage score were lower than that in the control group (p<0.05). MDA and MPO concentrations in the inflamed colonic tissues were decreased remarkably in the treated groups compared with that in the control group (p<0.05). But SOD level, IL-4 and IL-10 mRNA expression in the inflamed colonic tissues were obviously increased. ConclusionsIt is a potential path that protective effect of Si Shen Wan on impaired colonic mucosa rats with experimental colitis was accomplished by down-regulating the level of MDA and MPO, and up-regulating the level of SOD and the IL-4 and IL-10 mRNA expression in the colon mucosa.

Regulation of Sishen Wan on Bax/Bcl-2 mRNA, Fas/FasL in colonic tissue from rats with colitis

To evaluate therapeutic effect of Sishen Wan on experimental colitis, and explore its mechanism by expression of Bax/Bcl-2 mRNA, Fas/FasL in colonic tissue. Experimental colitis was induced by rectal administration of trinitrobenzene sulfonic acid (TNBS) dissolved in ethanol. The model animals were divided into four groups: the induced colitis but untreated group, the induced colitis groups treated with the high, middle, low dose of Sishen Wan, and the induced colitis group treated with salicylazosulfapyridine (SASP). After 10 day administration, the body weight, colonic wet weight, colonic weight index, colonic damage score and pathological change were evaluated, and the level of Fas and FasL by flow cytometry, Bax mRNA and Bcl-2 mRNA by reverse transcription polymerase chain reaction (RT-PCT). Compared with the model group, the colonic wet weight and colonic weight index were remarkably decreased in the middle dose of Sishen Wan group (P < 0.05). The colonic injury scores were significantly reduced after rats were treated with the three doses of Sishen Wan (P < 0.05). Representative restored features were observed including fewer inflammatory cellular infiltration and follicular hyperplasia, superficial and little ulcer with fibroplasia in colonic mucosa from the treated groups. The expression of Fas in the colonic mucosa was obviously down-regulated (P < 0.05) and the ratio of Bcl-2 mRNA/Bax mRNA was significantly up-regulated (P < 0.05) in the groups treated with the three doses of Sishen Wan. Sishen Wan might postpone colonic epithelium apoptosis or improve inflammatory cell apoptosis by regulating the expression of Fas/ FasL and Bax/Bcl-2 mRNA in colonic tissue, which is possible potential path to effectively treat experimental colitis by enema.

Characterization of the Cysteinyl Leukotriene 2 Receptor in Novel Expression Sites of the Gastrointestinal Tract

Cysteinyl leukotrienes (cysLTs: LTC₄, LTD₄, and LTE₄) are pro-inflammatory lipid molecules synthesized from arachidonic acid. They exert their actions on at least two cysLT receptors (CysLT₁R and CysLT₂R). Endothelial expression and activation of these receptors is linked to vasoactive responses and to the promotion of vascular permeability. Here we track the expression pattern of CysLT₂R in a loss-of-function murine model (CysLT₂R-LacZ) to neurons of the myenteric and submucosal plexus in the small intestine, colonic myenteric plexus, dorsal root ganglia, and nodose ganglion. Cysteinyl leukotriene (LTC₄/D₄) stimulation of colonic submucosal venules elicited a greater permeability response in wild-type mice. In a dextran sulfate sodium-induced colon inflammation model, the disease activity index and colonic edema (measured by wet:dry weights and submucosal thickness) were significantly reduced in knockout (KO) mice compared to controls. Tumor necrosis factor-α levels in colon tissue were significantly lower in KO mice; however, myeloperoxidase activity was similar in both the KO and wild-type groups. Finally, patch-clamp recordings of basal neuronal activity of colonic-projecting nociceptive neurons from dorsal root ganglia (T9-13) revealed significantly higher excitability in KO neurons compared to wild type. These results suggest that a lack of neuronal expression of CysLT₂R in the murine colonic myenteric plexus attenuates colitis disease progression via a reduction in inflammation-associated tissue edema and increases neuronal sensitivity to nociceptive stimuli.

Calcineurin inhibition by polaprezinc in rats with experimentally-induced colitis

Aims We investigated the therapeutic effect of polaprezinc (PZ), N-(3-aminopropionyl)- l-histidinato zinc, in rats with experimentally-induced colitis by focusing on calcineurin (CN) inhibition. CN plays a crucial role in T-cell activation and cytokine gene expression and is targeted by immunosuppressants such as cyclosporine and FK506. Main methods Colitis was induced into male Wistar rats by trinitrobenzene sulfonic acid and was treated with intrarectally administered PZ. The inflammation was assessed by the macroscopic damage score, colon wet weight, and proinflammatory mediator expression by RT-PCR analysis. Protein expression of calcineurin and the activation of its substrate, the nuclear factor of activated T cells (NFAT) transcription factor, were also studied. Calcineurin inhibition by PZ was investigated in in vitro experiments using colonic mucosa, purified calcineurin enzyme, and Jurkat T cells. Key findings CN was activated in the colitic mucosa; PZ treatment inhibited CN activation, the expression of proinflammatory cytokines in the mucosa, and thereby ameliorated the experimental colitis in rats. In in vitro experiments, PZ inhibited CN activity, NFAT activation, interleukin-2 expression, and the growth of Jurkat T cells. In the effective concentrations, PZ did not affect cell viability. Significance Our results suggest that PZ can be used as an immunosuppressive agent for the treatment of colitis through its inhibitory effect on CN activity.

Topical application of glycyrrhizin preparation ameliorates experimentally induced colitis in rats

To examine the efficacy of glycyrrhizin preparation (GL-p) in the treatment of a rat model of ulcerative colitis (UC). Experimental colitis was induced by oral administration of dextran sodium sulfate. Rats with colitis were intrarectally administered GL-p or saline. The extent of colitis was evaluated based on body weight gain, colon wet weight, and macroscopic damage score. The expression levels of pro-inflammatory cytokines and chemokines in the inflamed mucosa were measured by cytokine antibody array analysis. The effect of GL-p on myeloperoxidase (MPO) activity in the inflamed mucosa and purified enzyme was assayed. GL-p treatment significantly ameliorated the extent of colitis compared to sham treatment with saline. Cytokine antibody array analysis showed that GL-p treatment significantly decreased the expression levels of pro-inflammatory cytokines and chemokines, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α, cytokine-induced neutrophil chemoattractant-2, and monocyte chemoattractant protein-1 in the inflamed mucosa. Furthermore, GL-p inhibited the oxidative activity of mucosal and purified MPO. GL-p enema has a therapeutic effect on experimental colitis in rats and may be useful in the treatment of UC.