UC Model Research Articles

Ganluyin ameliorates DSS-induced ulcerative colitis by inhibiting the enteric-origin LPS/TLR4/NF-κB pathway

Ethnopharmacological relevanceUlcerative colitis (UC) is a chronic inflammatory bowel disease (IBD), that is associated with a significantly increased risk of colon cancer. As a classic traditional Chinese medicine, Ganluyin (GLY) has a long history as an anti-inflammatory medication, but its impacts on UC has not been established. Aim of the studyThis study aims to evaluate the protective effect and mechanism of GLY on a pathway involving enteric-origin lipopolysaccharide (LPS), toll-like receptor (TLR)4, and NF-κB in mice with dextran sulfate sodium (DSS)-induced UC. Materials and methodsAfter three weeks of intragastric administration of GLY, a UC model was induced in mice by administration of 4% DSS in drinking water for one week. The disease activity index (DAI) was measured, and histological staining was used to detect histopathological changes of colon. LPS content of the serum was measured by ELISA, and the expression of tight junction proteins and proteins related to TLR4/NF-κB pathway in colon were analyzed by immunohistochemistry or Western Blotting. The intestinal flora was analyzed by 16S rRNA sequencing. ResultsGLY improved the histological pathological changes of DSS-induced UC, as assessed by DAI, colonic mucosal damage, inflammatory cell infiltration, and goblet cell and mucus reduction. GLY also protected the intestinal mucosal barrier by increasing the expression of the tight junction proteins, occludin, claudin-1, and ZO-1 and by reducing the serum LPS content and decreasing the expression of TLR4, MyD88, NF-κB, IL-6, IL-1β, and TNF-α proteins in colon. Analyses of the intestinal flora showed that GLY restored the homeostasis of the intestinal flora through increases in the abundance of Firmicutes and decreases in the abundance of Proteobacteria and Bacteroidetes, which is associated with the production of LPS. ConclusionGLY might exert an anti-UC effect by improving the colonic mucosal barrier and inhibiting the enteric-origin LPS/TLR4/NF-κB inflammatory pathway, and restoring the homeostasis of the intestinal flora in UC mice. These discoveries lay a strong foundation for GLY as a UC treatment.

DIET AND ITS MICROENVIRONMENT INFLUENCE THE HEALTH OF COLON

Abstract Ulcerative colitis (UC) is an autoimmune chronic inflammatory disease of the large intestine that greatly increases risks of colorectal cancer. UC is an idiopathic inflammatory disorder of the colonic mucosa, which starts in the rectum and generally extends proximally in a continuous manner through part of, or the entire, colon. A Western diet, smoking, alcohol abuse, altered circadian rhythm and a dysbiotic microbiome is thought to induce UC. However, it is still not completely understood the factors in the diet that triggers UC. Identification of the factors associated with the diet that induce UC will lead to healthier lifestyle that prevents UC. In animal models we show that standard diet and diet rich in fiber positively influenced the health of the colon. A high fat high protein diet negatively regulated the health of the colon. A high fat high protein diet caused obesity in animals, paradoxically it also reduced the colon length significantly in these animals. Animals feeding on a high fat high protein had no energy in the fecal pellet demonstrating peinosis [hunger-like-condition; peina (Greek) hunger; osis (Greek) condition] of the colon. A high fat high protein diet also induced peripheral neuropathy in animals; whereas animals on a high fiber diet was healthy and had longer colon and no peripheral neuropathy. Bacteria of the family Erysipelotrichaceae that is related to inflammation-related disorders of the gastrointestinal tract is also high in animals fed with a high fat high protein diet; these animals also had low levels of valeric acid and propionic acid in the colon. Overall, a high fat high protein diet negatively impacts the health of the colon due to poor energy metabolism and its associated dysbiotic microbiome and metabolome. Currently, anti-inflammatory drugs and immune suppressors are prescribed for UC treatment, side-effects and the risk of opportunistic infections limit the quality of treatment. Hence, there is a need for a new class of biologics for efficient UC treatment. Bin1 is a risk factor in UC as determined by the increase in Bin1 expression in the colon of UC patients. In animal models, we showed that attenuation of BIN1 gene protected against UC. We have developed an innovative therapeutic strategy to treat UC that acts by correcting the leaky colonic epithelial barrier function that drives chronic inflammation in this setting. Specifically, we recently showed that administration of our anti-Bin1 monoclonal antibody (Bin1 mAb 99D) is sufficient to mimic the effect of genetic downregulation of Bin1 to preserve the normal colonic mucosa in an established mouse model of UC, in comparison to control-treated mice which exhibited severe lesions and inflammatory cell influx throughout the mucosa. A diet that positively influence the colon may be beneficial during immunotherapeutic treatment of ulcerative colitis.

Therapeutic effect of Patrinia villosa on TNBS-induced ulcerative colitis via metabolism, vitamin D receptor and NF-κB signaling pathways

Ethnopharmacological relevanceUlcerative colitis (UC) is a chronic inflammatory disease affecting the colon. Patrinia villosa Juss. (P.V) is an important traditional Chinese medicine widely used for more than 2000 years from ShenNongBenCaoJing, a famous ancient Chinese medicinal literary. P.V is often used in the treatment of UC, but the pathogenesis is not clear.Aim of the study: This study was designed to analysis the metabolic pathways and relevant mechanisms of P.V on UC rats induced by TNBS. Materials and methodsThe rat model of UC was established by 2,4,6-trinitrobenzene sulfonic acid (TNBS)/ethanol method. Three doses of P.V (21 g/kg, 43 g/kg, 64 g/kg) were administrated for 14 days. Disease activity index (DAI) scoring system and H&E staining were used to evaluate the efficacy. A method for simultaneous detection of 96 endogenous metabolic components was established by UPLC-MS. The method was used to detect the metabolites in serum and liver of rats with UC induced by TNBS. PLS-DA and Metaboanalyst were used to analyze the main metabolic pathways involved in the treatment of UC. The contents of IL-1β, TNF-α and IL-6 in the colonic homogenate of rats were detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of VDR, NF-κB, p-NF-κB, NLRP3 and caspase-1 in colon tissues of rats were detected by the method of Western blot. ResultsDAI scoring system and H&E staining indicated that P.V have the obvious therapeutic effect on UC induced by TNBS as a dosage-dependent manner. 36 potential biomarkers in serum and 26 potential biomarkers in liver were found in positive and negative ion mode of UPLC-MS, which significantly affected Glycine, serine and threonine metabolism, Glycerophospholipid metabolism, Purine metabolism, Histidine metabolism, Alanine, aspartate, and glutamate metabolism, Arginine and proline metabolism in serum, and significantly affected Purine metabolism, Lycine, serine and threonine metabolism, Glutathione metabolism, Glyoxylate and dicarboxylate metabolism in the liver. The contents of pro-inflammatory cytokines and receptors related to NF-κB signaling axis of model group were significantly higher than those of the control group, compared with the model group, their contents of the P.V group were significantly decreased (p < 0.01). Compared with the model group, the expression of NF-κB, p-NF-κB, NLRP3 and caspase-1 in colon tissues of the rats in P.V group were significantly decreased (p < 0.01). The expression of VDR in model group were significantly reduced compared to that in the control group, compared with the model group, the expression of VDR in P.V group were significantly increased (p < 0.01). ConclusionP.V has an obvious therapeutic effect on UC induced by TNBS by regulating the energy metabolism, amino acid metabolism, purine metabolism, bile acid metabolism and lipid metabolism. P.V exerts anti-inflammatory effect by impacting bile acid levels, activating VDR, and inhibiting the overactivation of NF-κB signaling pathways.

Neferine, a natural alkaloid from Nelumbo nucifera, ameliorates experimental chronic ulcerative colitis in mice

ObjectiveUlcerative colitis (UC), a chronic inflammatory disease, is closely associated with the development of colorectal cancer. Neferine is a natural alkaloid that possesses many therapeutic effects. Our previous studies showed that neferine showed a significant protective effect on the acute ulcerative colitis mouse model. Here, the protective role of neferine on chronic UC was investigated. MethodsA chronic mice UC model was established by employing dextran sodium sulfate (DSS) and neferine was orally administrated. The disease activity index (DAI) scores were calculated. The levels of pro-inflammatory mediators were determined by ELISA kits. The pathological changes were evaluated by H&E staining. Protein expressions were detected by immunohistochemistry and Western blotting. ResultsNeferine treatment significantly decreased the DAI scores, ameliorated pathological injuries, and reduced the levels of myeloperoxidase (MPO), IL-1β, IL-6, and TNF-α. Furthermore, the expressions of NF-κB p65, receptor-interacting protein kinase 1 and 3 (RIP1 and RIP3), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in colon tissues were suppressed by neferine. In addition, the decreased expression of caspase-8 and IκBα were restored by neferine. ConclusionNeferine protects chronic UC by regulating the secretion of pro-inflammatory cytokines and inhibiting the NF-κB signaling pathway.

Proteomic analysis of lysine acetylation reveals that metabolic enzymes and heat shock proteins may be potential targets for DSS-induced mice colitis

BackgroundResearch on acetylation modification and its modification sites will be of great significance for revealing the mechanism of disease and developing new targeted medicines. In this study, we aim to construct a complete atlas of acetylome in the DSS-induced ulcerative colitis mice model (UC model) MethodsA high-resolution mass spectrometry-based quantitative approach was employed to identify lysine-acetylated proteins and acetylation sites. Bioinformatics analysis and in vitro experiments verified anti-inflammatory effects of HSP90B1-K142ac. Results2597 acetylation events and 1914 sites were quantified, highlighting 140 acetylation site changes in the colitis colon tissue. 91 acetylation sites in 75 proteins were up-regulated, and 49 acetylation sites in 39 proteins were down-regulated in the UC models. The differentially acetylated proteins mainly consisted of non-histone proteins located in the cytoplasm and mitochondria. KEGG and protein-protein interaction networks analysis showed that the differentially acetylated proteins were enriched in the TCA cycle, fatty acid metabolism, and protein processing in the endoplasmic reticulum. 68% of the differentially metabolized enzymes have a down-regulated trend in acetylation levels. The acetylation level of lysine 142 in HSP90B1 was found to be obvious in the UC colon, and point mutation of HSP90B1-K142ac would result in the decreasing secretion of TNF-α and IL-2 in LPS-stimulated cultured cells. ConclusionOur work built a complete atlas of acetylome and revealed the potential role of metabolic enzymes and heat shock proteins in DSS-induced colitis.

PD37-08 PHOSPHATIDYLDIGLYCEROL-BASED THERMOSENSITIVE LIPOSOMES SHOW SUPERIOR THERAPEUTIC EFFICACY AGAINST MUSCLE-INVASIVE BLADDER CANCER IN VIVO

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology I (PD37)1 Sep 2021PD37-08 PHOSPHATIDYLDIGLYCEROL-BASED THERMOSENSITIVE LIPOSOMES SHOW SUPERIOR THERAPEUTIC EFFICACY AGAINST MUSCLE-INVASIVE BLADDER CANCER IN VIVO Iris Brummelhuis, Michiel Simons, Lars Lindner, Simone Kort, Sytse de Jong, Martin Hossann, Alfred Witjes, and Egbert Oosterwijk Iris BrummelhuisIris Brummelhuis More articles by this author , Michiel SimonsMichiel Simons More articles by this author , Lars LindnerLars Lindner More articles by this author , Simone KortSimone Kort More articles by this author , Sytse de JongSytse de Jong More articles by this author , Martin HossannMartin Hossann More articles by this author , Alfred WitjesAlfred Witjes More articles by this author , and Egbert OosterwijkEgbert Oosterwijk More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002047.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Recommended treatments for muscle-invasive bladder cancer (MIBC) are associated with substantial morbidity. Hyperthermia (HT) triggered drug release from phosphatidyldiglycerol-based thermosensitive liposomes (DPPG2-TSL) might prevent surgical bladder removal and toxicity from systemic chemotherapy. We aimed to assess the efficacy of DPPG2-TSL encapsulated doxorubicin (DOX; DPPG2-TSL-DOX) combined with bladder HT, compared to free systemic and intravesical DOX, in a syngeneic orthotopic rat urothelial carcinoma model. METHODS: A total of 191 female Fischer F344 rats were used. Bladder tumors were initiated by inoculation of AY-27 cells and tumor bearing rats were selected with cystoscopy and semi-randomized over treatment groups. At day 5 and 8, animals were treated with 2 mg/kg DOX in different treatment modalities: intravenous (iv) DPPG2-TSL-DOX with HT, iv free DOX without HT, intravesical DOX without HT, intravesical DOX with HT or no treatment (control group), respectively. Animals were euthanized at day 14 and complete tumor response was assessed by histopathological evaluation. RESULTS: Treatment with iv DPPG2-TSL-DOX with HT resulted in the highest rate of animals with complete tumor response of 70%, which was significantly better than iv free DOX (18%, p=0.02) and no treatment (0%, p=0.001), and non-significantly better than intravesical DOX with (43%, p=0.35) or without HT (50%, p=0.41; Figure 1). All rats receiving intravesical DOX with HT (7/7) and 24% (5/21) of rats treated with DPPG2-TSL-DOX containing the same DOX dose with HT had to be euthanized before day 14 because of >15% body weight loss, while some rats showed dilated ureters (3/7 and 4/21) and urine retention (2/7 and 1/21, respectively).Figure 1. Tumor stage as determined by histopathological evaluation per group. Green colored parts of bars represent T1 tumors, and purple T2. CONCLUSIONS: Treatment with DPPG2-TSL-DOX combined with intravesical HT outperformed systemic and intravesical DOX in a syngeneic orthotopic rat UC model. There might be a role for DPPG2-TSL encapsulating chemotherapeutics in treatment of MIBC in the future. Source of Funding: This study was partially sponsored by Thermosome © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e658-e658 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Iris Brummelhuis More articles by this author Michiel Simons More articles by this author Lars Lindner More articles by this author Simone Kort More articles by this author Sytse de Jong More articles by this author Martin Hossann More articles by this author Alfred Witjes More articles by this author Egbert Oosterwijk More articles by this author Expand All Advertisement Loading ...

Systems pharmacology and transcriptomics reveal the mechanisms of Sanhuang decoction enema in the treatment of ulcerative colitis with additional Candida albicans infection

BackgroundUlcerative colitis (UC) is an important inflammatory phenotype in bowel disease (IBD), which is caused by multiple potential factors, including fungal dysbiosis. Candida albicans (C. albicans) was confirmed to be an important factor promoting the occurrence and development of UC. Sanhuang decoction (SHD) has been used for UC therapy in China for thousand of years, although its core active constituents and pharmacological mechanism remain undefined.MethodsIn this work, a murine model of UC with C. albicans colonization was established with dextran sodium sulfate (DSS) and C. albicans intragastric administration. The major bioactive constituents and potential mechanism of SHD against UC with fungal dysbiosis were comprehensively examined by combining systems pharmacology and in vivo transcriptomics.ResultsSHD attenuated C. albicans burden, reduced DAI, increased mucosal integrity and relived systemic inflammation in UC mice. Systems pharmacology analysis identified 9 core bioactive ingredients and 45 hub targets of SHD against UC. Transcriptomics analysis confirmed 370 differentially expressed genes (DEGs) after SHD treatment, which were mainly enriched in inflammatory and immune response related signaling pathways. Toll-like receptor and PI3K-Akt signaling pathway were screened out as the candidate targets involved in the action of SHD on fungal dysbiosis-associated UC, which were consistent with the findings in systems pharmacology. The expression of TLR4, IL-1β, NF-κB, PI3K and Akt proteins were stimulated by C. albicans, and partially reversed by SHD in UC mice.ConclusionThese findings suggested SHD could be a candidate for the treatment of fungal dysbiosis-associated UC via TLR4-NF-κB and PI3K-Akt signaling pathways.

Importance of Telomere Shortening in the Pathogenesis of Ulcerative Colitis: A New Treatment From the Aspect of Telomeres in Intestinal Epithelial Cells.

Ulcerative colitis [UC] is a chronic inflammatory disease of the colon with frequent relapses. Telomere shortening in intestinal epithelial cells has been reported in severe or longstanding cases. However, its influence on UC pathogenesis remains unelucidated. To this end, we evaluated telomere shortening using a long-term organoid inflammation model that we had originally established. A UC model using human colon organoids was established to assess telomere changes chronologically. MST-312 was used for the telomerase inhibition assay. The potential of telomerase activators as a novel UC treatment was evaluated with an in vitro model, including microarray analysis, and histological changes were assessed using xenotransplantation into mouse colonic mucosa. Our UC model reproduced telomere shortening in vitro, which was induced by the continuous suppression of telomerase activity via P53. MST-312-based analysis revealed that telomere shortening was involved in the pathogenesis of UC. Madecassoside [MD] improved the telomere length of the UC model and UC patient-derived organoids, which further promoted cell proliferation in vitro and improved the graft take-rate of xenotransplantation. Moreover, histological analysis revealed that MD induced normal crypt structure with abundant goblet cells. This study is the first to reveal the mechanism and importance of telomere shortening in the pathogenesis of UC. MD could be a novel candidate for UC treatment beyond endoscopic mucosal healing.

Suppressing Kv1.3 Ion Channel Activity with a Novel Small Molecule Inhibitor Ameliorates Inflammation in a Humanised Mouse Model of Ulcerative Colitis.

Background and AimsThe potassium channel Kv1.3 is a potentially attractive therapeutic target in T cell-mediated inflammatory diseases, as the activity of antigen-activated T cells is selectively impeded by Kv1.3 inhibition. In this study, we examined Kv1.3 as a potential therapeutic intervention point for ulcerative colitis [UC], and studied the efficacy of DES1, a small-molecule inhibitor of Kv1.3, in vitro and in vivo.MethodsKv1.3 expression on T cells in peripheral blood mononuclear cells [PBMCs] isolated from donors with and without UC was examined by flow cytometry. In biopsies from UC patients, Kv1.3-expressing CD4+ T cells were detected by flow cytometry and immunohistochemistry. In vitro, we determined the ability of DES1 to inhibit anti-CD3-driven activation of T cells. In vivo, the efficacy of DES1 was determined in a humanised mouse model of UC and compared with infliximab and tofacitinib in head-to-head studies.ResultsKv1.3 expression was elevated in PBMCs from UC patients and correlated with the prevalence of TH1 and TH2 T cells. Kv1.3 expression was also detected on T cells from biopsies of UC patients. In vitro, DES1 suppressed anti-CD3-driven activation of T cells in a concentration-dependent manner. In vivo, DES1 significantly ameliorated inflammation in the UC model and most effectively so when PBMCs from donors with higher levels of activated T cells were selected for reconstitution. The efficacy of DES1 was comparable to that of either infliximab or tofacitinib.ConclusionInhibition of Kv1.3 [by DES1, for instance] appears to be a potential therapeutic intervention strategy for UC patients.

Regulation of IL-6/STAT-3/Wnt axis by nifuroxazide dampens colon ulcer in acetic acid-induced ulcerative colitis model: Novel mechanistic insight

AimUlcerative colitis (UC) is a common intestinal problem characterized by the diffusion of colon inflammation and immunity dysregulation. Nifuroxazide, a potent STAT-3 inhibitor, exhibits diverse pharmacological properties. The present study aimed to elucidate a novel anti-colitis mechanism of nifuroxazide against the acetic acid-induced UC model. MethodsRats were grouped into control (received vehicle), UC (2 ml of 5% acetic acid by intrarectal infusion), UC plus sulfasalazine (100 mg/kg/day, P.O.), UC plus nifuroxazide (25 mg/kg/day, P.O.), and UC plus nifuroxazide (50 mg/kg/day, P.O.) and lasted for 6 days. ResultsThe present study revealed that nifuroxazide significantly reduced UC measures, hematological changes, and histological alteration. In addition, treatment with nifuroxazide significantly down-regulated serum CRP as well as the colonic expressions of MPO, IL-6, TNF-α, TLR-4, NF-κB-p65, JAK1, STAT-3, DKK1 in a dose-dependent manner. Besides, our results showed that the colonic Wnt expression was up-regulated with nifuroxazide treatment. In a dose-dependent manner, nifuroxazide markedly alleviated acetic acid-induced cellular infiltration and improved ulcer healing by increasing intestinal epithelial cell regeneration. SignificanceOur results collectively indicate that nifuroxazide is an effective anti-colitis agent through regulation of colon inflammation and proliferation via modulation IL-6/STAT-3/Wnt signaling pathway.

6,7-Dihydroxy-2,4-Dimethoxyphenanthrene from Chinese Yam Peels Alleviates DSS-Induced Intestinal Mucosal Injury in Mice via Modulation of the NF-κB/COX-2 Signaling Pathway.

In this study, we evaluated the protective effect and molecular mechanism of a dominant phenanthrene, (6,7-dihydroxy-2,4-dimethoxyphenanthrene, CYP4), from Chinese yam peels on intestinal epithelial integrity. Three doses of Chinese yam phenolic extract (CYPE) and Chinese yam phenanthrene 4 (CYP4) were administered to BALB/c mice for 7 days before dextran sulfate sodium (DSS) treatment, with berberine hydrochloride as a positive control (PC). Results showed that both disease activity indexes (DAIs), histological damage score (HDS) and survival rate in DSS mice, were improved with preintervention of CYPE and CYP4, which exhibited better efficiency than PC. Further studies showed that administration of CYP4 downregulated the oxidative stress-associated factors, MPO and NO, and improved tight junction protein occludin. Besides, the CYP4 treatment substantially downregulated the caspase-3 expression and the apoptosis rate of intestinal epithelial cells. In addition, the CYP4 treatment ameliorated the production of inflammatory cytokines including TNF-α, IFN-γ, IL-10, and IL-23 in the colon. Furthermore, the protein expression of ERK1/2, NF-κB p65, pNF-κB, and COX-2 was suppressed in CYE4 groups as compared with that in model control (MC). These findings suggested that CHP4 could effectively inhibit the activation of NF-κB/COX-2 in an experimental UC model in vivo. It was demonstrated for the first time that CYPE and CYP4 protected intestinal mucosa from damage and prevented DSS-induced colitis in mice. CYP4 was one of the active principles obligatory for the biological effect of Chinese yam in protecting intestinal health. These findings indicated that CYP4 might be a promising and useful approach for treatment of UC in humans.

Suppression of ELF4 in ulcerative colitis predisposes host to colorectal cancer

SummaryUlcerative colitis (UC) is a chronic inflammatory bowel disease, characterized by relapsing and remitting colon mucosal inflammation. For patients suffering from UC, a higher risk of colon cancer has been widely recognized. Here, we found that Elf4−/− mice developed colon tumors with 3 cycles of dextran sulfate sodium salt (DSS) treatment alone. We further showed that ELF4 suppression was prevalent in both patients with UC and DSS-induced mice models, and this suppression was caused by promoter region methylation. ELF4, upon PARylation by PARP1, transcriptionally regulated multiple DNA damage repair machinery components. Consistently, ELF4 deficiency leads to more severe DNA damage both in vitro and in vivo. Oral administration of montmorillonite powder can prevent the reduction of ELF4 in DSS-induced colitis models and lower the risk of colon tumor development during azoxymethane (AOM) and DSS induced colitis-associated cancer (CAC). These data provided additional mechanism of CAC initiation and supported the “epigenetic priming model of tumor initiation”.

Dental pulp stem cells overexpressing hepatocyte growth factor facilitate the repair of DSS-induced ulcerative colitis

BackgroundUlcerative colitis (UC) is a chronic and recurrent disease without satisfactory treatment strategies. Dental pulp stem cell (DPSC) transplantation has been proposed as a potential therapy for UC. This study aimed to investigate the therapeutic effects of the rat hepatocyte growth factor (HGF) gene transduced into DPSCs for UC.MethodsThe therapeutic effects of HGF-DPSCs transplanted intravenously into a rat model of UC induced by 5% dextran sulphate sodium (DSS) were compared with the other treatment groups (LV-HGF group, DPSCs group and GFP-DPSCs group). Immunofluorescence and immunohistochemistry were used to observe the localization and proliferation of HGF-DPSCs at the site of colon injury. The expression levels of inflammatory factors were detected by real-time quantitative PCR (RT-PCR) and western blotting. The oxidative stress markers were detected by ELISA. DAI scores and body weight changes were used to macroscopically evaluate the treatment of rats in each group.ResultsImmunofluorescence and immunohistochemistry assays showed that HGF-DPSCs homed to colon injury sites and colocalized with intestinal stem cell (ISC) markers (Bmi1, Musashi1 and Sox9) and significantly promoted protein expression (Bmi1, Musashi1, Sox9 and PCNA). Anti-inflammatory cytokine (TGF-β and IL-10) expression was the highest in the HGF-DPSCs group compared with the other treatment groups, while the expression of pro-inflammatory cytokines (TNF-α and INF-γ) was the lowest. Additionally, the oxidative stress response results showed that malondialdehyde (MDA) and myeloperoxidase (MPO) expression decreased while superoxide dismutase (SOD) expression increased, especially in the HGF-DPSCs group. The DAI scores showed a downward trend with time in the five treatment groups, whereas body weight increased, and the changes were most prominent in the HGF-DPSCs group.ConclusionsThe study indicated that HGF-DPSCs can alleviate injuries to the intestinal mucosa by transdifferentiating into ISC-like cells, promoting ISC-like cell proliferation, suppressing inflammatory responses and reducing oxidative stress damage, which provides new ideas for the clinical treatment of UC.

Network pharmacology and pharmacokinetics integrated strategy to investigate the pharmacological mechanism of Xianglian pill on ulcerative colitis

BackgroundUlcerative colitis (UC) is a chronic inflammatory bowel disease with high morbidity, which leads to poor quality of life. The Xianglian pill (XLP) is a classical Chinese patent medicine and has been clinically proven to be an effective treatment for UC. PurposeThe pharmacological mechanism of the key bioactive ingredients of XLP for the treatment of UC was investigated by a network pharmacology and pharmacokinetics integrated strategy. Study design and methodsNetwork pharmacology was used to analyze the treatment effect of nine quantified XLP ingredients on UC. Key pathways were enriched and analyzed by protein-protein interaction and Kyoto Encyclopedia of Genes and Genomes analyses. The effect of XLP on Th17 cell differentiation was validated using a mouse model of UC. The binding of nine compounds with JAk2, STAT3, HIF-1α, and HSP90AB1 was assessed using molecular docking. A simple and reliable ultra-high-performance liquid chromatography-tandem mass spectrometry method was developed for the simultaneous quantification of nine ingredients from XLP in plasma and applied to a pharmacokinetic study following oral administration. ResultsNine compounds of XLP, including coptisine, berberine, magnoflorine,berberrubine, jatrorrhizine, palmatine, evodiamine, rutaecarpine, and dehydrocostus lactone, were detected. Network pharmacology revealed 50 crossover genes between the nine compoundsand UC. XLP treats UC mainly by regulating key pathways of the immune system, including Th17 cell differentiation, Jak-Stat, and PI3K-Akt signaling pathways. An in vivo validation in mice found that XLP inhibits Th17 cell differentiation by suppressing the Jak2-Stat3 pathway, which alleviates mucosal inflammation in UC. Molecular docking confirmed that eight compounds are capable of binding with JAk2, HIF-1α, and HSP90AB1, further confirming the inhibitory effect of XLP on the Jak2-Stat3 pathway. Moreover, apharmacokinetic study revealed that the nine ingredients of XLP are exposed in the plasma and colon tissue, which demonstrates its pharmacological effect on UC. ConclusionThis study evaluates the clinical treatment efficacy of XLP for UC. The network pharmacology and pharmacokinetics integrated strategy evaluation paradigm is efficient in discovering the key pharmacological mechanism of herbal formulae.

Mogrol, an aglycone of mogrosides, attenuates ulcerative colitis by promoting AMPK activation

BackgroundUlcerative colitis (UC) is a non-specific chronic inflammatory disease. The incidence of UC in China has been increasing in recent years. Mogrol is an aglycone of mogrosides. Studies have shown that mogrosides have anti-oxygenation, anti-inflammatory, and laxative effects as well as other biological activities. PurposeTo investigate the beneficial effects of mogrol on UC and identify its underlying mechanisms. Study designWe used the dextran sodium sulphate (DSS)-induced UC model in mice, TNF-α-damaged NCM460 colonic epithelial cells, macrophage cells THP-M stimulated with lipopolysaccharide (LPS) / adenosine triphosphate (ATP) and compound C (an AMPK inhibitor) to confirm the key role of AMPK (AMP-activated protein kinase) activation. MethodsHistological evaluation, immunohistochemical staining, Western blot analysis, immunofluorescence assay and quantitative real time-PCR were used in the study. ResultsOral administration of mogrol (5 mg/kg/daily) in vivo significantly attenuated pathological colonic damage, inhibited inflammatory infiltration and improved the abnormal expression of NLRP3 inflammasome in colonic mucosa via the AMPK and NF-κB signaling pathways. In vitro, mogrol protected against intestinal epithelial barrier dysfunction by activating AMPK in TNF-α-treated NCM460 cells and inhibited the production of inflammatory mediator in LPS-stimulated THP-M cells. Furthermore, mogrol's effects were reversed by compound C intervention in DSS-induced UC model. ConclusionMogrol exerts protective effects in experimental UC and inhibits production of inflammatory mediators through activation of AMPK-mediated signaling pathways.

Copaifera malmei Harms leaves infusion attenuates TNBS-ulcerative colitis through modulation of cytokines, oxidative stress and mucus in experimental rats

Ethnopharmacological relevanceEthnobotanical studies show that the infusion of the leaves from Copaifera malmei Harms (Fabaceae) has been utilized in the Brazilian traditional medicine to treat provocative and gastrointestinal diseases, among others. Recently, our research team has shown that an infusion extract of the leaves of C. malmei has a strong antiulcer activity and its oral use gives no indications of toxicity. Aim of the studyThe aim of the study is to evaluate the anti-inflammatory intestinal effect of an infusion extract from the leaves of Copaifera malmei (IECm) in an animal model of ulcerative colitis induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). Materials and methodsAcute intestinal inflammation was induced in male Wistar rats by TNBS in 20% EtOH (0.25 mL). IECm was administered by oral gavage (for 72, 48, 24 and 2 h) preceding the induction of ulcerative colitis. The colon damage and degree of inflammation were evaluated by morphological observation scores and colon weight. The improved colonic mucosal injury, oxidative stress and inflammatory response were assessed by histopathological investigation and by estimating myeloperoxidase (MPO) activity, malondialdehyde (MDA) and glutathione (GSH) levels and tumor necrosis factor (TNF), interleukin 1β (IL1-β), IL-17 and IL-10 colon tissue concentrations. The histopathological changes were done on the colon tissues by hematoxylin and eosin and Periodic Acid-Schiff staining were utilized to measure the mucus. ResultsPre-treatment (25, 100 and 400 mg/kg) with IECm altogether diminished the intestinal inflammation prompted by TNBS in rats by diminishing colonic score by 69.12% (p < 0.01), 19.87% (p < 0.05) and 67.60% (p < 0.01), individually. Improvement of colonic mucosal injury by treatment with IECm was shown by a decline in MPO activity at dosages 25 and 400 mg/kg by 67.98% and 59.68% (p < 0.001), MDA levels 64.80% and 80.00% (p < 0.01) and an expansion in GSH content at all portions (62.53%, 53.38% and 81.20% p < 0.05) compared with vehicle control group. IECm additionally prevention of intestinal inflammation as confirm by decreased cytokine levels, for example, TNF (31.26%, p < 0.05, 50.68% and 45.95%, p < 0.01), IL1-β (56.41%, 58.83% and 56.65%, p < 0.001), IL-17 (51.66%, p < 0.001, 22.23%, p < 0.05 and 49.67%, p < 0.001) and increased the IL-10 levels at 25 and 400 mg/kg (57.13%, p < 0.01 and 35.83%, p < 0.05) respectively. Histopathological examination of the colon tissue displayed recovery of ulcerative colitis of IECm treated animals by reducing leukocyte infiltrate, epithelial, submucosal and muscular layer damages and maintaining mucus production. ConclusionThese findings revealed that IECm was effective and possess anti-colitic activities in a rodent model of UC and can be useful in inflammatory bowel diseases (IBD). The pre-treatment with IECm decreased intestinal inflammation by reducing macroscopical and microscopical colon injury. In addition, the present study demonstrated that IECm ameliorates TNBS-colitis by promoting antioxidant effect, modulation of cytokines release and restauration of mucus production. The study reinforces the traditional use of the Copaifera malmei leaves infusion to inflammatory gastrointestinal disorders and makes IECm a potential herbal medicine for the treatment of IBD.

The IITM Model: A Simple and Expressive Model for Universal Composability

The universal composability paradigm allows for the modular design and analysis of cryptographic protocols. It has been widely and successfully used in cryptography. However, devising a coherent yet simple and expressive model for universal composability is, as the history of such models shows, highly non-trivial. For example, several partly severe problems have been pointed out in the literature for the UC model. In this work, we propose a coherent model for universal composability, called the IITM model (“Inexhaustible Interactive Turing Machine”). A main feature of the model is that it is stated without a priori fixing irrelevant details, such as a specific way of addressing of machines by session and party identifiers, a specific modeling of corruption, or a specific protocol hierarchy. In addition, we employ a very general notion of runtime. All reasonable protocols and ideal functionalities should be expressible based on this notion in a direct and natural way, and without tweaks, such as (artificial) padding of messages or (artificially) adding extra messages. Not least because of these features, the model is simple and expressive. Also the general results that we prove, such as composition theorems, hold independently of how such details are fixed for concrete applications. Being inspired by other models for universal composability, in particular the UC model and because of the flexibility and expressivity of the IITM model, conceptually, results formulated in these models directly carry over to the IITM model.

Joint State Composition Theorems for Public-Key Encryption and Digital Signature Functionalities with Local Computation

In frameworks for universal composability, complex protocols can be built from sub-protocols in a modular way using composition theorems. However, as first pointed out and studied by Canetti and Rabin, this modular approach often leads to impractical implementations. For example, when using a functionality for digital signatures within a more complex protocol, parties have to generate new verification and signing keys for every session of the protocol. This motivates to generalize composition theorems to so-called joint state (composition) theorems, where different copies of a functionality may share some state, e.g., the same verification and signing keys. In this paper, we present a joint state theorem which is more general than the original theorem of Canetti and Rabin, for which several problems and limitations are pointed out. We apply our theorem to obtain joint state realizations for three functionalities: public-key encryption, replayable public-key encryption, and digital signatures. Unlike most other formulations, our functionalities model that ciphertexts and signatures are computed locally, rather than being provided by the adversary. To obtain the joint state realizations, the functionalities have to be designed carefully. Other formulations proposed in the literature are shown to be unsuitable. Our work is based on the IITM model. Our definitions and results demonstrate the expressivity and simplicity of this model. For example, unlike Canetti’s UC model, in the IITM model no explicit joint state operator needs to be defined and the joint state theorem follows immediately from the composition theorem in the IITM model.

Oral Delivery of β-Lactoglobulin-Nanosphere-Encapsulated Resveratrol Alleviates Inflammation in Winnie Mice with Spontaneous Ulcerative Colitis

Resveratrol (RES) is a nutraceutical with promising anti-inflammatory properties for the treatment of inflammatory bowel diseases (IBD). However, the clinical effectiveness of resveratrol as an oral anti-inflammatory agent is hindered by its extremely poor solubility and poor stability. In this study, we encapsulated resveratrol in β-lactoglobulin (BLG) nanospheres and systematically analyzed their formulation parameters in vitro followed by a thorough in vivo anti-inflammatory testing in a highly specialized spontaneous murine UC model (Winnie mice model). Complexation of resveratrol with BLG increased the aqueous solubility of resveratrol by ≈1.7 times with 10% w/w loading. Additionally, the in vitro dissolution of resveratrol from the particles was found to be higher compared to resveratrol alone, resulting in >90% resveratrol dissolution in ∼8 h. The anti-inflammatory activity of resveratrol was examined for the first time in Winnie mice, a mouse model that closely represents the clinical signs of IBD. At a 50 mg/kg oral dose for 2 weeks, BLG-RES significantly improved both % body weight and disease activity index (DAI), compared to free resveratrol in Winnie mice. Importantly, histological evaluations revealed a similar trend with striking improvement in the pathology of the colon via an increase in goblet cell numbers and recovery of colonic epithelium. BLG-RES significantly increased the expression level of cytokine interleukin-10 (Il10), which confirms the reduction in inflammation potentially because of the increased dissolution and stability of resveratrol by complexation with BLG. This comprehensive study demonstrates the effectiveness of biocompatible nanomaterials such as BLG in oral delivery of poorly soluble anti-inflammatory molecules such as resveratrol in the treatment of IBD.

EXPERIMENTAL ULCERATIVE COLITIS: TGF-Β AS A DIAGNOSTIC MARKER

Previous studies stated that TGF-β is a pleiotropic cytokine, abundant in mammalian intestine, regulates multiple cellular functions as a suppressor of the immune response, cell proliferation, and oncogenesis. Studyaims: Here, we investigated the value of TGF-β as a diagnostic marker of ulcerative colitis disease (UC) in experimental UC model. Materials& Methods: Acute & chronic UC was induced in mice by using (3% and 1.5% respectively) DSS in drinking water. The induction of UC was confirmed by histopathological examination. Proteins and mRNA expression of both TGF-β and TNF-α were analyzed by western blot and real time PCR respectively. Complete blood count (CBC), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR). Results: A marked elevation occurred in proteins and mRNA expression of both TGF-β and TNF-α in UC tissues in acute and chronic groups vs. healthy group. Furthermore, significant elevation in both CRP and ESR of diseased groups was recorded in comparison to normal group . Blood count revealed marked leukocytosis, thrombocytosis and marked decrease in red blood cell counts with severe anemia in both acute and chronic UC groups compared to healthy control group . Conclusion: Increased levels of TGF-β protein and mRNA expression in UC disease might have a diagnostic value side by side with inflammatorybiomarkers (TNF-α, CRP & ESR), leukocytosis, thrombocytosis, bloody diarrhea and histopathological changes.