Intrarectal Administration Research Articles

Protective effect of 6-paradol in acetic acid-induced ulcerative colitis in rats

BackgroundUlcerative colitis is a gut inflammatory disorder due to altered immune response to gut microbiome, with interplay of environmental and genetic factors. TNF-α activates inflammatory response through a cascade of immune responses, augmenting pro-inflammatory mediators and proteases, activating chemotaxis, and infiltration of inflammatory cells, leading to ulceration and haemorrhage through cytotoxic reactive oxygen species. 6-Paradol, a dietary component in several plants belonging to the Zingiberaceae family, has shown anti-inflammatory and antioxidant activities. Current study evaluates the effect of 6-paradol in amelioration of ulcerative colitis in rats for the first time.Methods6-Paradol (95% purity) was obtained from seeds of Aframomum melegueta. Rats were divided randomly into six groups (n = 8). Group one was administered normal saline; group two was treated with the vehicle only; group three, sulfasalazine 500 mg/kg; and groups four, five, and six, were given 6-paradol (50, 100, 200, respectively) mg/kg orally through gastric gavage for 7 days. Colitis was induced on 4th day by intrarectal administration of 2 ml acetic acid (3%), approximately 3 cm from anal verge. On 8th day, rats were sacrificed, and distal one-third of the colon extending proximally up to 4 cm from anal orifice was taken for biochemical and gross examination. Two centimetres of injured mucosal portion was taken for histopathological investigations. SPSS (ver.26) was used for statistical analysis.ResultsColonic and serum glutathione (GSH) levels decreased, while colonic and serum malondialdehyde (MDA), colonic myeloperoxidase (MPO) activity, serum interleukin-6 (IL-6), serum tumour necrosis factor-α (TNF-α) levels, and colon weight to length ratio were increased significantly in the colitis untreated group compared to normal control. Treatment with 6-paradol considerably improved all these parameters, especially at a dose of 200 mg/kg (p < 0.001), revealing non-significant differences with sulfasalazine 500 mg/kg and normal control (p = 0.998). Sulfasalazine and 6-paradol in a dose dependent manner also markedly reversed mucosal oedema, atrophy and inflammation, cryptic damage, haemorrhage, and ulceration. There were non-significant differences between low and medium doses and between medium and high doses of 6-paradol for IL-6 and serum MDA levels.Conclusion6-Paradol demonstrated protection against acetic acid-induced ulcerative colitis, probably by anti-inflammatory and antioxidant actions.

The Therapeutic Potential of Time-Restricted Fasting on Experimental Ulcerative Colitis

Aim: Intermittent fasting have been reported to have beneficial effect, in that it improves gut microbiota and lowers inflammation. This research is, however, targeted at evaluating the healing effects of fasting on ulcerative colitis in rats.&#x0D; Study Design: A total of eighteen Wistar rats were used for this study, and were divided into three major groups; animals that were neither induced with colitis nor fasted (group 1), animals with colitis and were allowed to fast (group 2), animals with colitis but were not allowed to fast (group 3).&#x0D; Place and duration of study: Department of Physiology, Ladoke Akintola University of Technology, Ogbomoso, Oyo State, between March, 2020 and July, 2020&#x0D; Methodology: The weight of the 18 animals used was 180 ± 20 g. Colitis was induced by a single dose of intra-rectal administration of 1mL/100g body weight of 6% acetic acid. Animals in group 1 served as control animals. Animals in group 2 were only given access to food between 4:00 p.m and 6:30 p.m, whereas animals in group 3 were given food and water ad libitum. Animals were sacrificed ten days post colitis induction. Colonic levels of Tumor necrotic factor-alpha (TNFα), Glutathione (GSH) concentration, Superoxide dismutase (SOD), Catalase (CAT) and Myeloperoxidase (MPO) activities were measured. The blood glucose level in the animals was also recorded by the use of a glucometer.&#x0D; Results: There was a reduction in the concentration of TNFα and GSH, an increase in CAT and SOD activities in the colitis animals that were allowed to fast when compared with colitis animals that were not allowed to fast, ten days post colitis induction. There was also the lowering of blood sugar level, all signifying the beneficial effect of fasting on chemically-induced colitis. Thus, intermittent fasting helped the animals to heal from chemically-induced colitis.

Prophylactic effect of aquatic extract of stevia on acetic acid induced-ulcerative colitis in male rats: a possible role of Nrf2 and PPARγ.

Ulcerative colitis (UC) is a non-specific intestinal inflammatory disease. Several studies demonstrated that inflammation and oxidative stress play significant role in the pathogenesis of this disease. This study aimed to determine the protective effect and possible mechanism by which stevia affects the course of experimentally induced colitis. Male rats were received stevia 20, 40, 80mg/kg/day before induction of colitis by intra-rectal administration of 2mL of 4% acetic acid, AA. Macroscopic and histopathological examination of the colon were done. Colonic content of catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH), myeloperoxidase (MPO) and thiobarbituric acid reactive substances (TBARS) activities and serum levels of interleukin (IL)1- β and tumor necrosis factor (TNF)-α were assessed. Real time-PCR (RT-PCR) was done to determine the expression of NF-κB, Nrf2 and PPARγ genes. Spontaneous contraction and effects of increasing concentrations of acetylcholine and stevia have been studied on the isolated colonic segments. Stevia ameliorated colitis not only histopathologically but also it decreased the level of TNF-α, IL-1β, TBARS, MPO and the expression of NF-κB which were significantly increased in the AA group. The concentration of GSH, SOD, CAT and expression of Nrf2 and PPARγ were significantly increased with stevia. Moreover, stevia showed a relaxant effect on the colonic contractility which was increased in AA group. These all effects of stevia were more prominent with its highest dose. Our results explored that, stevia acts protectively against UC by its anti-inflammatory and antioxidant properties which mediated by up-regulation of Nrf2 and PPARγ with downregulation of NF-κB. We suggest that stevia has the potential for treatment of chronic inflammatory diseases, such as UC.

Comparison of pharmacokinetics of intragastrically and intrarectally administered levetiracetam tablets in healthy non-epileptic dogs.

Canine status epilepticus (CSE) is characterized by epileptic seizures that are longer than 5 min or more than one seizure with incomplete recovery. Currently, diazepam suppositories are generally prescribed for CSE. Levetiracetam (LEV) is one of the newest antiepileptic drugs currently available. This study compared the pharmacokinetics of intragastric and intrarectal administration in oral formula of LEV in four healthy beagles as a reference data when the owner administers levetiracetam to dogs by himself at home. Blood for measuring plasma LEV concen- trations was collected 0, 30, 60, 90, 120, 240, 360, and 540 min after LEV administration. The time to reach the maximum plasma concentration (Tmax) was markedly shorter with intra- rectal administration (45±26 min) than with intragastric administration (270±99 min). Intrarectal administration of LEV tablets could be an effective option for treating canine seizures although it might be a limit for treating CSE because the absorption rate is not fast enough.

Elucidation of colon-protective efficacy of diosgenin in experimental TNBS-induced colitis: inhibition of NF-κB/IkB-α and Bax/Caspase-1 signaling pathways.

ANOVA: Analysis of variance; 5-ASA: 5-aminosalicylic acid; Bax: Bcl-2-associated X protein; COX-2: Cyclooxygenase-2; DAI: Disease Activity Index; DMSO: Dimethyl sulfoxide; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; GSH: Glutathione; HP: Hydroxyproline; IAEC: International Animal Ethics Committee; IBD: Inflammatory Bowel Disease; IBS: Inflammatory Bowel Syndrome; IL's: Interleukin's; IFN-γ: Interferon-gamma; IκBα: nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor-alpha; iNOs: Inducible nitric oxide synthase; LTB4: Leukotriene B4; MDA: Malondialdehyde; MPO: Myeloperoxidase; NO: Nitric Oxide; NF-κB: Nuclear Factor-κB; ROS: Reactive Oxygen Species; SOD: Superoxide Dismutase; TNBS: Trinitrobenzene Sulfonic Acid; TNF-α: Tumor necrosis factor-α.

Comparison of the tolerability and efficacy of intra-rectal lidocaine gel with peri-prostatic nerve block as anaesthetic techniques for prostate biopsy

BackgroundProstate biopsy is a painful procedure. However, over the years, varied opinions exist among urologists about the methods of achieving optimal pain relief for the procedure. The purpose of this study was to compare the tolerability of administration and the efficacy of intra-rectal lidocaine gel with peri-prostatic nerve block for this procedure.MethodsFrom June 2016 to June 2017, 110 patients who met the study criteria were randomized to receive either intra-rectal lidocaine gel (Group 1) or an apical peri-prostatic nerve block (Group 2) for prostate biopsy. Their level of pain perception during the anaesthesia administration and the prostate biopsy was assessed using the Numeric Rating Scale. Other biopsy-related complications and patient satisfaction were also evaluated. Grouped data were compared using student t test and Chi-square with p < 0.05 considered significant.ResultsThe pain score during the administration of anaesthesia was 1.6 ± 1.9 and 3.7 ± 2.1 for the intra-rectal lidocaine gel and peri-prostatic nerve block groups, respectively (p = 0.001). However, during the prostate biopsy, the pain score was 6.8 ± 2.2 and 2.9 ± 1.9 for the intra-rectal lidocaine gel and peri-prostatic nerve block groups, respectively (p = 0.001). There was a significantly lower rate of satisfaction (45.3% versus 86.8%, p = 0.001) in the lidocaine gel group.ConclusionsAlthough intra-rectal lidocaine gel administration is better tolerated by patients, a peri-prostatic nerve block is more effective and provides better patient satisfaction than intra-rectal lidocaine gel when used as anaesthesia for prostate biopsy.

Risperidone attenuates acetic acid-induced colitis in rats through inhibition of TLR4/NF-kB signaling pathway

Aim The purpose of the present study is to explore the anti-inflammatory potential of risperidone in acetic acid-induced rat colitis through inhibition of TLR4/NF-kB pathway. Methods Acute colitis induction was done by intra-rectal administration of 2 mL of 4% diluted acetic acid solution. Two h after colitis induction, dexamethasone (2 mg/kg) as standard drugorrisperidone (2, 4 and 6 mg/kg) were administered orally to wistar rats for five consecutive days. 24 h after the last treatment, animals were sacrificed by cervical dislocation. Macroscopic and microscopic damage evaluation was done. Biochemical and ELISA methods were used to assess myeloid peroxidase (MPO) enzyme activity and tumor necrosis factor-α (TNF-α) level respectively. Moreover, immunohistochemistry (IHC) was performed to detect the expression of TLR4 and pNF-kBproteins. Results Dexamethasone (2 mg/kg) or risperidone (2, 4 and 6 mg/kg) improved acetic acid-induced macroscopic (p < .001) and microscopic lesions. Additionally, risperidone (2, 4 and 6 mg/kg) inhibited the activity of MPO and TNF-α (p < .01, p < .001) in the colon tissue compared to acetic acid group. Furthermore, bothdexamethasone and risperidone (2, 4 and 6 mg/kg) significantly reduced acetic acid-induced expression of TLR4and pNF-kB proteins (p < .05, p < .01, p < .001). Conclusion The anti-inflammatory effect of risperidone on acetic acid-induced colitis in rats may involve inhibition of TLR4 and NF-kB signaling pathway.

The Effects of Dietary Glycine on the Acetic Acid-Induced Mouse Model of Colitis.

Inflammatory bowel disease, a gut disease that is prevalent worldwide, is characterized by chronic intestinal inflammation, such as colitis, and disorder of the gut microbiome. Glycine (Gly) is the simplest amino acid and functions as an anti-inflammatory immune-nutrient and intestinal microbiota regulator. This study aimed at investigating the effect of Gly on colitis induced in mice by intrarectal administration of 5% acetic acid (AA). Bodyweight and survival rates were monitored, and colonic length and weight, serum amino acid concentrations, intestinal inflammation-related gene expression, and colonic microbiota abundances were analyzed. The results showed that Gly dietary supplementation had no effect on the survival rate or the ratio of colonic length to weight. However, Gly supplementation reversed the AA-induced increase in serum concentrations of amino acids such as glutamate, leucine, isoleucine, and valine. Furthermore, Gly inhibited colonic gene expression of interleukin- (IL-) 1β and promoted IL-10 expression in colitis mice. Gly supplementation also reversed the AA-induced reduction in the abundance of bacteria such as Clostridia, Ruminococcaceae, and Clostridiales. This change in the intestinal microbiota was possibly attributable to the changes in colonic IL-10 expression and serum concentrations of valine and leucine. In sum, Gly supplementation regulated the serum concentrations of amino acids, the levels of colonic immune-associated gene expression, and the intestinal microbiota in a mouse model of colitis. These findings enhance our understanding of the role of Gly in regulating metabolism, intestinal immunity, and the gut microbiota in animals afflicted with colitis.

Attempts to induce tolerance to Trichostrongylus colubriformis infection in sheep.

The possibility of manipulating the immune response in lambs to the gastrointestinal nematode Trichostrongyluscolubriformis to reduce production losses associated with infection was investigated. In a series of four experiments, attempts to immunize sheep via the mucosal route to modify the immune response and induce mucosal tolerance are outlined. Initially, a proof of concept study was conducted with lambs being injected with multiple doses of a somatic Tcolubriformis antigen without an adjuvant in the rectal submucosa and subsequently challenged with Tcolubriformis L3 larvae. This was followed by a dose-response study comparing different antigen doses to identify the optimum dose of the nematode antigen for successful induction of mucosal tolerance. The final two studies were conducted to determine the larval stage specificity of the parasite antigen and the most suitable site of delivery required to stimulate mucosal tolerance. In the proof of concept study, lambs either received repeated injections in the rectal submucosa at 3×weekly intervals with 15µg of L3, 11µg of L4 and 21µg of immature adult (L5) somatic Tcolubriformis antigens (ANT) or not (INF) prior to infection with Tcolubriformis. In the dose-rate study, antigen dose rates of 100%, 50%, 10%, 1% or 0% of the antigen concentration used in the proof of concept study were compared while the larval stage study compared antigen from either L3, L4, L5 stages or combination of all (COMB) and the route of administration study compared antigen delivery into either the rectal submucosa (RE) or sub-cutaneous injection (SC). During infection, lamb growth was improved by antigen treatment between days 21 and 42 in the proof of concept study (P=.009), for groups 10%, 50% and 100% in the dose-rate study (P<.05 for all) and in RE in the route of administration study with no improvement observed in the larval stage study. No differences in faecal egg counts were observed (P>.05 for all). Parasite-specific IgA and IgE showed a dose-response (the dose-rate study), were not affected by larval stage (the larval stage study) and were greater in RE than SC (the route of administration study). IL-4 production following lymphocyte stimulation was greatest in COMB (the larval stage study) and RE (the route of administration study). Although antigen treatment improved performance, this was inconsistent and appeared to stimulate immunity rather than induce tolerance. Combined larval stages were more efficient than individual stages, and intra-rectal administration was more effective than sub-cutaneous.

Histone H3K9 methylation regulates chronic stress and IL-6-induced colon epithelial permeability and visceral pain.

Chronic stress is associated with activation of the HPA axis, elevation in pro-inflammatory cytokines, decrease in intestinal epithelial cell tight junction (TJ) proteins, and enhanced visceral pain. It is unknown whether epigenetic regulatory pathways play a role in chronic stress-induced intestinal barrier dysfunction and visceral hyperalgesia. Young adult male rats were subjected to water avoidance stress±H3K9 methylation inhibitors or siRNAs. Visceral pain response was assessed. Differentiated Caco-2/BBE cells and human colonoids were treated with cortisol or IL-6±antagonists. Expression of TJ, IL-6, and H3K9 methylation status at gene promoters was measured. Transepithelial electrical resistance and FITC-dextran permeability were evaluated. Chronic stress induced IL-6 up-regulation prior to a decrease in TJ proteins in the rat colon. The IL-6 level inversely correlated with occludin expression. Treatment with IL-6 decreased occludin and induced visceral hyperalgesia. Chronic stress and IL-6 increased H3K9 methylation and decreased transcriptional GR binding to the occludin gene promoter, leading to down-regulation of protein expression and increase in paracellular permeability. Intrarectal administration of a H3K9 methylation antagonist prevented chronic stress-induced visceral hyperalgesia in the rat. In a human colonoid model, cortisol decreased occludin expression, which was prevented by the GR antagonist RU486, and IL-6 increased H3K9 methylation and decreased TJ protein levels, which were prevented by inhibitors of H3K9 methylation. Our findings support a novel role for methylation of the repressive histone H3K9 to regulate chronic stress, pro-inflammatory cytokine-mediated reduction in colon TJ protein levels, and increase in paracellular permeability and visceral hyperalgesia.

The Anti-Inflammatory Effect of Pistacia Lentiscus in a Rat Model of Colitis.

IntroductionThe mastic tree (Pistacia lentiscus), belonging to the Anacardiaceae family, has anti-inflammatory and antioxidant properties. This study aims to assay the anti-inflammatory effects of mastic in rats with colitis.MethodsForty-eight male Sprague-Dawley rats were randomly divided into six groups of control, colitis without treatment, colitis with mastic (400 mg/kg/daily) administered orally or intra-rectally, colitis with prednisolone (5 mg/kg of body weight), and colitis with sesame oil for seven successive days. Treatment effects were evaluated by determining cytokines (TNFα, IL6) and myeloperoxidase (MPO) activity, macroscopic scores, and histopathological parameters. The results of each group are compared with colitis without treatment group.ResultsAfter administering sesame oil, the MPO level was reduced significantly compared to colitis without the treatment group (P=0.025). The mastic oil (400 mg/kg orally) administration was effective in reducing colitis severity through the reduction in the total colitis index (p=0.046) after 7 days. The Intra-rectal administration of mastic decreased TNF-α significantly, similar to prednisolone and control groups compared to the colitis without treatment group (p=0.024). The IL-6 did not change in the mastic and sesame oil groups.ConclusionAccording to our results, mastic and sesame oil have anti-inflammatory properties, suggesting that they could be used as natural sources to lessen the ulcerative colitis inflammation.

Bupropion Ameliorates Acetic Acid-Induced Colitis in Rat: the Involvement of the TLR4/NF-kB Signaling Pathway.

Inflammatory bowel disease composed of ulcerative colitis and Crohn's disease is a disorder that may involve entire gastrointestinal tract. Its pathogenesis is mainly an immune-mediated inflammation. Recently, it has been indicated that bupropion possesses anti-inflammatory properties; hence, the objective of this experiment is the investigation of the anti-inflammatory influence of bupropion on colonic lesions that emerged following the intrarectal administration of acetic acid. Thirty-six male Wistar rats were allocated randomly into six groups, including control, acetic acid, dexamethasone (2mg/kg), and bupropion (40, 80, and 160mg/kg). Colitis was induced by intrarectal administration of acetic acid in all study groups except control group, and animals were treated by oral administration of dexamethasone and bupropion. While macroscopic and microscopic lesions were observed after colitis induction, administration of dexamethasone and bupropion 160mg/kg led to the remarkable improvement in lesions. In addition, the expression of TLR4 and NF-ĸB was decreased after colitis induction; however, treatment with dexamethasone (2mg/kg) and bupropion (160mg/kg) resulted in a significant decrease in their expression. Regarding biochemical factors, following colitis induction, TNF-α level and MPO activity were increased; nevertheless, dexamethasone (2mg/kg) and bupropion (160mg/kg) decreased the TNF-α and MPO activity. In conclusion, bupropion exerts anti-inflammatory influence through suppressing the TLR4 and NF-ĸB expression in the rat model of acute colitis.

Targeting IL-10, ZO-1 gene expression and IL-6/STAT-3 trans-signaling by a combination of atorvastatin and mesalazine to enhance anti-inflammatory effects and attenuates progression of oxazolone-induced colitis.

Ulcerative colitis (UC) is a chronic inflammatory disease characterized by diffused inflammation of the colon and rectum mucosa. The pathogenesis of UC is multifactorial, and the exact underlying mechanisms remain poorly understood. This study aims to investigate the effect of mesalazine and atorvastatin combination in enhancing anti-inflammatory effects and attenuates progression of oxazolone colitis in rats. In the present study, male albino rats (N=60) were divided into six groups (10 rats each), the first two groups served as normal control and a control saline group. Colitis was induced by intra-rectal administration of oxazolone in the 5th and 7th days after pre-sensitization. Then, rats were divided into untreated group, groups treated with mesalazine or atorvastatin or their combination. Colitis was assessed by colon length, body weight, and incidence of diarrhea, rectal bleeding, and histopathology of colon tissue. Colon tissues were used for measuring interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), IL-13, signal transducer and activator of transcription-3 (STAT-3), myeloperoxidase activity (MPO), reduced glutathione(GSH), and tissue expression of IL-10, tight junction protein zonula occludens (ZO-1), and caspase-3 genes. The combination therapy significantly attenuated progression of UC by decreasing incidence of diarrhea, rectal bleeding, weight loss, IL-13, IL-6, TNF-α, STAT-3, caspase-3, and MPO activity and significantly increased IL-10, ZO-1, colon length, and GSH content, and these effects were more superior to single drugs. These findings showed that combination therapy was able to ameliorate progression of UC and enhance anti-inflammatory effects possibly by restoring IL-10 and ZO-1 levels and limiting IL-6/STAT-3 trans-signaling.

Effect of recombinant serine protease from adult stage of Trichinella spiralis on TNBS-induced experimental colitis in mice

Inflammatory bowel disease (IBD), which includes ulcerative colitis (UC) and Crohn’s disease (CD), is a chronic autoimmune disease. At present, worms and their products has been shown to have protective effects on immune-mediated diseases. Therefore, we aimed to investigate the effect of the recombination Trichinella spiralis (T. spiralis, Ts) adult serine protease-like protein rTs-ADSp-7 on a 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced CD mouse model. Colitis was induced by intrarectal administration of a TNBS solution. The disease activity index (DAI), which included weight loss, diarrhoea, and bloody stool, was measured. Colon segments were stained with haematoxylin and eosin (H.E.) for histopathological score. Cytokine release in the serum was analysed by meso scale discovery (MSD). Cytokine release in the colon was detected by ELISA. Splenocytes were separated, and the cytokine profiles of Th1 (IFN-γ), Th2 (IL-4), Th17 (IL-17A) and Treg cells were analysed by flow cytometry. Our result showed that rTs-ADSp-7 reduced the clinical disease activity of TNBS-induced colitis in mice. In addition, we found that rTs-ADSp-7 reduced the production of Th1- and Th17-related cytokines while upregulating the expression of Th2- and Treg-related cytokines in TNBS-induced colitis mice. rTs-ADSp-7 also increased the population of Th2 and Treg cells in TNBS-induced colitis mice. rTs-ADSp-7 alleviated the severity of TNBS-induced colitis while balancing the CD4+ T cell immune response. rTs-ADSp-7 has therapeutic potential for colitis treatment and can be used as a helminth-derived protein therapy for CD or other Th1 immunity-mediated diseases.

The effect of Hericium erinaceum on the prevention of chemically induced experimental colitis in rats.

Background/AimsThe aim of this study is to investigate the effects of the Hericium erinaceum on an experimental colitis model.MethodsTwenty-four Wistar albino were included in this study. Rats were divided into three groups. Group 1 (n = 8) was sham group. Group 2 is the group of chemically induced by intrarectal administration of trinitrobenzene sulfonic acid (TNBS) resulting in colitis. Group 3 (n = 8) is the group that was treated 7 days before and 7 days after with H. erinaceum resulting in colitis. The activity of colitis was evaluated macroscopically and microscopically in rats. In other words, nitric oxide (NO) levels, malondialdehyde (MDA), interleukin 6 (IL-6), nuclear factor-kappa B (NF-κB) and, tumor necrosis factor-α (TNF-α) in addition to the myeloperoxidasem (MPO) activities was determined.ResultsThe rate of TNBS-induced colitis caused to increase the level of MDA activities meaningfully in the colitis group than the control group. The results indicated that MDA (p = 0.001), NO (p = 0.001), IL-6 (p = 0.001), MPO (p = 0.878), TNF-α (p = 0.001), and NF-κB levels of treatment group decreased in the blood and colon tissues because of the H. erinaceum treatment when compared to the colitis group. H. erinaceum treatment was related to the declining of MDA, NF-κB, NO, IL-6, and TNF-α levels.ConclusionsH. erinaceum had a positive effect on the colitis by reducing oxidative damage in blood and tissue.

Sacral nerve stimulation ameliorates colonic barrier functions in a rodent model of colitis.

The mucosal barrier damage is recognized as one of the key factors in the pathogenesis of colitis. While sacral nerve stimulation (SNS) was reported to have therapeutic potential for colitis, its mechanisms of actions on colonic permeability remained largely unknown. In this study, colitis was induced by intrarectal administration of TNBS in rats. Five days later, they were treated with SNS or sham-SNS for 10days. The effects of SNS on colonic permeability were assessed by measuring the expression of tight-junction proteins involved in regulating permeability and the FITC-dextran test. The mechanism of actions of SNS was investigated by studying the function of the enteric nervous system (ENS) cells and analyzing the autonomic nervous system. SNS decreased the disease activity index, microscopic and macroscopic scores, myeloperoxidase activity, and pro-inflammatory cytokines (TNF-α, IL-6). SNS increased the expression of Zonula Occludens-1, Occludin, Claudin-1, and Junctional adhesion molecule-A in the colon tissue. The FITC-dextran test showed that the colonic permeability was lower with SCS than sham-SNS. SNS increased ChAT, pancreatic polypeptide, and GDNF and reduced norepinephrine NGF, sub-P, and mast cell overactivation in the colon tissue. Concurrently, SNS increased acetylcholine in colon tissues and elevated vagal efferent activity. SNS ameliorates colonic inflammation and enhances colonic barrier function with the proposed mechanisms involving the increase in parasympathetic activity and modulation of the activity of the ENS and immune system, including mast cells.

Arthrocen, an avocado‐soy unsaponifiable agent, improves acetic acid‐induced colitis in rat by inhibition of NF‐kB signaling pathway

The goal of the current study was to evaluate the anti-inflammatory effect of Arthrocen against acetic acid-induced colitis in rats. Acute inflammation was produced through intrarectal administration of 2ml diluted acetic acid (4%) solution. All interventions were carried out for 5days after colitis induction. Arthrocen was administered orally at doses of 30, 60, and 120mgkg-1 day-1 . Then, macroscopic and microscopic studies were performed. Myeloperoxidase (MPO) activity and tumor necrosis factor-α (TNF-α) activity were measured by biochemical and ELISA methods, respectively. Immunohistochemistry was done to investigate the expression of pNF-κB. The results of this study demonstrated that Arthrocen reduced macroscopic and microscopic damage compared to the acetic acid group. Furthermore, Arthrocen decreased the activity of MPO and TNF-α as well as the protein expression of pNF-kB in rat colon tissue. The results of the current study revealed the anti-inflammatory activity of Arthrocen in acetic acid mediated colon inflammation through suppressing the NF-κB pathway. PRACTICAL APPLICATIONS: Inflammatory bowel disease (IBD) is an immune-mediated chronic relapsing disorder affecting the gastrointestinal tract (GIT) characterized by chronic bowel inflammation. A plant-based dietary supplement containing avocado and soy unsaponifiable extracts in a ratio of 1:2 is known as Arthrocen. Arthrocen can be used as a complementary drug beside current drugs in clinical trials for the treatment of IBD.

Mucosal Thrombin Alters Gut Microbiota Biofilms Structure And Promote Dispersion Of Bacteria With Aggressive Behavior

Alterations of gut microbiota have been implicated in a broad variety of intestinal diseases. The mechanisms whereby this may occur remains elusive. Gut mucosal microbiota is naturally organized as a polymicrobial biofilm, separated from the intestinal epithelium by a sterile mucus layer. We recently discovered that intestinal epithelium releases active thrombin, which plays a key role in segregation of intestinal biofilms from host tissue. Furthermore, we detected an upregulation of active thrombin in inflamed human patients and in models of colitis.ObjectivesOur study objective was to determine whether exposure to high thrombin, such as this occurring during inflammation, will alter commensal microbiota biofilms and promote the dispersion of bacteria predisposed to damage the intestinal epithelium.MethodsMicrobiota extracted from healthy human colon biopsies were seeded into the Calgary Biofilm Device and polystyrene coupons to develop, a multispecies anaerobic biofilm. Biofilms were exposed to various concentrations of thrombin (10 to 1000 Unit/ml). Dispersed bacteria released from thrombin‐treated biofilms were collected and their composition was assessed by 16S sequencing. These bacteria were apically exposed to human epithelial monolayers on transwells (Caco2 and HT29MTX). Adhesion (90 minutes), invasion (gentamicin assay, 3 hours) and translocation (4 hours) to basolateral side was quantified by plating on agar. Transwells were processed for fluorescent in situ hybridization for bacteria staining and phalloidin antibody for host cell cytoskeleton. Motility phenotype of biofilmdispersed bacteria was assessed on soft agarose gels (swarming and swimming). Mice (B6) were treated intracolonically with thrombin (5U per day for 10 days) or boiled thrombin (similar dose). Rats (Wistar) were treated with TNBS to induce colitis, and were treated for 3 days intracolonically with dabigatran (thrombin inhibitor, 1 μg/kg) or vehicle.ResultsBiofilm‐dispersed bacteria from thrombin‐treated biofilms attached more importantly to the epithelial monolayers compared to untreated biofilm. 3D reconstruction images confirmed such thrombin‐induced phenotype. Thrombin alters swarming and swimming motility in soft agarose gel. 16S analysis further precise the specific composition of biofilm‐dispersed bacteria after thrombin exposure. In mice, intrarectal administration of thrombin caused alterations of gut microbiota biofilms structure (16S sequencing) and organization ( in situ imaging of gut microbiota). During colitis in rats, local inhibition of thrombin activity prevented gut microbiota biofilms alterations associated with colitis (in situ imaging of gut microbiota and 16S analysis).ConclusionsThese data suggest that high concentration of thrombin released at gut mucosal surface during inflammation alters gut biofilm organization and modifies the phenotype of biofilm‐dispersed bacteria, which were able to invade and cross the epithelial barrier, thus increasing their likelihood to trigger inflammatory flares.

Andrographolide sulfonate ameliorates chronic colitis induced by TNBS in mice via decreasing inflammation and fibrosis

Inflammatory bowel disease could result in diarrhea and abdominal pain, as well as potential complications such as tissue fibrosis. The therapeutic effect of andrographolide sulfonate on acute murine experimental colitis induced by 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) has been confirmed. In the study here, chronic colitis triggered by repeated intrarectal administration of TNBS was established and the effect of andrographolide sulfonate was examined. Repeated TNBS administration induced substantial mice death, which was significantly decreased by andrographolide sulfonate treatment. The elevation of inflammatory cytokines including IL-6, IL-17A, TNF-α as well as IFN-γ in colonic tissues levels were decreased after administration of andrographolide sulfonate. Next, CD4+ T cell and macrophage infiltration was found to descend. The subset of pathogenic CD4+ T cell subset including CD4+IFN-γ+ (Th1) and CD4+IL-17A+ (Th17) were also suppressed by andrographolide sulfonate. Further, the restrain of p38 and p65 activation were also observed after andrographolide sulfonate administration. Finally, TNBS-induced colonic epithelial damage as well as fibrosis were significantly mitigated by andrographolide sulfonate. Based on the results got here, we can make a conclusion that andrographolide sulfonate could decrease inflammation and epithelial damage as well as fibrosis thus ameliorating chronic colitis in mice. Our study suggest the possible use of andrographolide sulfonate for chronic colitis treatment in clinical.

Piper umbellatum L. (Piperaceae): Phytochemical profiles of the hydroethanolic leaf extract and intestinal anti-inflammatory mechanisms on 2,4,6 trinitrobenzene sulfonic acid induced ulcerative colitis in rats

Ethnopharmacological relevancePiper umbellatum L. leaves, commonly found in the Amazon, Cerrado and Atlantic rain forest regions of Brazil, are widely used as a traditional medicine to treat gastrointestinal disorders and inflammation, among others diseases. Also, previous scientific studies demonstrated that P. umbellatum has gastroprotective and anti-inflammatory activity. AimTo investigate the phytochemical profiles and the intestinal anti-inflammatory effect of the hydroethanolic extract of P. umbellatum (HEPu) leaf on ulcerative colitis in rats. Materials and methodsIn this study, the chemical composition of HEPu was analyzed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography coupled to mass spectrometry (LC/MS). Also, this work studied the effects of HEPu on ulcerative colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS, 30 mg/mL in 20% ethanol) by intrarectal administration in rats. Simultaneously, animals were pre-treated orally with HEPu (30, 100 and 300 mg/kg), mesalazine (500 mg/kg), or vehicle. At the end of the experimental period, clinical signs of ulcerative colitis were evaluated by determination of weight loss, gross appearance, ulcer area and histological changes. Reduced glutathione (GSH), lipoperoxides (MDA) and nitric oxide (NO) levels, and superoxide dismutase (SOD), myeloperoxidase (MPO) and catalase (CAT) activities were determined in colon tissues. Also, pro-inflammatory mediators such as tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL- 1β) were quantified by immunoassay on the surface of fluorescent-coded magnetic beads (Luminex MagPix System). ResultsGC-MS analysis showed the presence of 17 different phytochemical compounds in the HEPu. LC/MS analyses revealed the presence of compounds in HEPu as protocatechuic acid, ferulic acid, kaempferol, rosmarinic acid, apigenin and ursolic acid. Treatment with HEPu significantly ameliorated weight loss, macroscopic damage, ulcerated area and histopathological changes such as sub-mucosal edema, cell infiltration, ulceration and necrosis (p < 0.001). Furthermore, HEPu (30, 100, and 300 mg/kg, p.o) inhibited the levels of oxidative parameters, such as MPO (49%, 53%, and 62%, p < 0.001), NO (20%, 19%, 22%, p < 0.01), and MDA (75%, 83%, 70%, p < 0.001), whereas increased the antioxidant activities such as SOD (208%, 192%, 64%, p < 0.001), GSH (94%, 75%, 49%, p < 0.01), and CAT (92%, 69%, 108%, p < 0.01). The extract also inhibited the pro-inflammatory cytokines TNF-α (81%, 85%, 85%, p < 0.001) and IL-1β (95%, 79%, 89%, p < 0.001) levels. ConclusionTogether, these results revealed that P. umbellatum L. is a promising source of metabolites to be used in the treatment of inflammatory bowel disease.