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The ABCC subfamily contains thirteen members. Nine of these transporters are called multidrug resistance proteins (MRPs). The MRPs have been associated with developing ulcerative colitis (UC). This study aimed to evaluate the ABCC expression in UC patients and its role in a dextran sulfate sodium (DSS)-induced colitis mice model under 5-aminosalicylates or methylprednisolone treatment and compared with control without inflammation. DSS-induced colitis mice were treated with 5-aminosalicylates (50 mg/kg 24 h) or methylprednisolone (2 mg/kg 24 h). Human rectal biopsies were obtained from UC patients. The abcc-relative mRNA levels and protein expression were determined by RT-PCR and immunohistochemistry. abcc4, abcc5, and abcc6 mRNA levels were significantly increased in DSS-induced colitis compared to the other groups. The 5-aminosalicylate treatment dramatically increased the abcc2 and abcc3 mRNA levels vs. control. Methylprednisolone treatment increased abcc1 vs. DSS-induced colitis and colitis treated with 5-aminosalicylate. Immunohistochemical analysis revealed down-regulation of ABCC1/ABCC2/ABCC5/ABCC7 in mice colitis vs. control. Treatment with 5-aminosalicylate restored ABCC5 levels, while methylprednisolone restored ABCC2/ABCC5/ABCC7 in colitis mice at similar control levels. Relative mRNA levels of mrp1-5 were increased in active UC patients vs. control. ABCC2/ABCC4/ABCC7 were conspicuously expressed in the mucosa of 5-aminosalicylate and/or methylprednisolone-treated UC patients, while ABCC2/ABCC4/ABCC5/ABCC7 in submucosa, ABCC1/ABCC5/ABCC7 in muscular, and ABCC1/ABCC4/ABCC5/ABCC7 in serosa were expressed vs. controls. This is the first report about the differential up-regulation of the ABCC subfamily gene and protein expression in DSS-induced colitis under aminosalicylates or methylprednisolone treatment.

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Inflammatory bowel diseases (IBD) are idiopathic disorders characterized by chronic gastrointestinal inflammation. Given conventional therapies' adverse effects and clinical failures, novel approaches are being investigated. Recent studies have highlighted the role of specialized pro-resolving lipid mediators (SPMs) in the active resolution of chronic inflammation. In this regard, omega-3 fatty acid-derived Resolvin D2 (RvD2) appears to play a protective role in the pathophysiology of IBD. Therefore, we characterized the RvD2 pathway and its receptor expression in the intestinal mucosa of experimental colitis induced by dextran sulfate sodium. We also evaluated the preventive impact of an omega-3-enriched diet and the therapeutic efficacy of RvD2 compared with anti-TNF-α treatment. We found an increase in TNFα and IL22 expression and decreased levels of enzymes involved in RvD2 biosynthesis, such as PLA2, 15-LOX, 5-LOX, and its receptor GPR18 in experimental colitis. Omega-3 supplementation reduced the Disease Activity Index (DAI), weight loss, colonic shortening, and inflammation. These results and the increased IL-10 transcriptional levels after RvD2 treatment suggest that this mediator attenuated experimental colitis. These results enhance our understanding of the molecular mechanisms involved in the exacerbated inflammatory response present in experimental colitis and suggest that RvD2 and its omega-3 precursor offer a promising therapeutic approach for IBD.

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Colorectal cancer CRC remains one of the leading causes of cancer-related deaths worldwide, with chronic intestinal inflammation identified as a major risk factor. Notably, the tumor suppressor TP53 undergoes mutation at higher rates and earlier stages during human inflammation-driven colon tumorigenesis than in sporadic cases. We investigated whether deleting Trp53 affects inflammation-induced tumor growth and the expression of Lgr5+ cancer stem cells in mice. We examined azoxymethane (AOM)/dextran sodium sulfate (DSS)-induced colon tumorigenesis in wild-type Trp53 (+/+), heterozygous (+/-), and knockout (-/-) mice. Trp53-/- mice showed increased sensitivity to DSS colitis and earlier accelerated tumorigenesis with 100% incidence. All groups could develop invasive tumors, but knockouts displayed the most aggressive features. Unlike wild-type CRC, knockouts selectively showed increased populations of Lgr5+ colon cancer stem-like cells. Trp53 loss also boosted laminin, possibly facilitating the disruption of the tumor border. This study highlights how Trp53 deletion promotes the perfect storm of inflammation and stemness, driving colon cancer progression. Trp53 deletion dramatically shortened AOM/DSS latency and improved tumor induction efficiency, offering an excellent inflammation-driven CRC model.

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Ulcerative colitis has been associated with psychological distress and an aberrant immune response. The immunomodulatory role of systemic cytokines produced during experimental intestinal inflammation in tonic immobility (TI) defensive behavior remains unknown. The present study characterized the TI defensive behavior of guinea pigs subjected to colitis induction at the acute stage and after recovery from intestinal mucosa injury. Moreover, we investigated whether inflammatory mediators (tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-8, IL-10, and prostaglandins) act on the mesencephalic nucleus, periaqueductal gray matter (PAG). Colitis was induced in guinea pigs by intrarectal administration of acetic acid. The TI defensive behavior, histology, cytokine production, and expression of c-FOS, IBA-1, and cyclooxygenase (COX)-2 in PAG were evaluated. Colitis reduced the duration of TI episodes from the first day, persisting throughout the 7-day experimental period. Neuronal c-FOS immunoreactivity was augmented in both columns of the PAG (ventrolateral (vlPAG) and dorsal), but there were no changes in IBA-1 expression. Dexamethasone, infliximab, and parecoxib treatments increased the duration of TI episodes, suggesting a modulatory role of peripheral inflammatory mediators in this behavior. Immunoneutralization of TNF-α, IL-1ß, and IL-8 in the vlPAG reversed all effects produced by colitis. In contrast, IL-10 neutralization further reduced the duration of TI episodes. Our results reveal that peripherally produced inflammatory mediators during colitis may modulate neuronal functioning in mesencephalic structures such as vlPAG.

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Yerba Mate (YM) (Ilex paraguariensis) is a natural herbal supplement with a well-described anti-inflammatory capacity and beneficial effects in different inflammatory contexts such as insulin resistance or obesity. However, whether YM could improve other inflammatory conditions such as colitis or the immune cell population that can be modulated by this plant remains elusive. Here, by using 61 male and female C57BL/6/J wild-type (WT) mice and the dextran sodium sulfate (DSS)-induced acute colitis model, we evaluated the effect of YM on colitis symptoms and macrophage polarization. Our results showed that the oral administration of YM reduces colitis symptoms and improves animal survival. Increasing infiltration of anti-inflammatory M2 macrophage was observed in the colon of the mice treated with YM. Accordingly, YM promoted M2 macrophage differentiation in vivo. However, the direct administration of YM to bone marrow-derived macrophages did not increase anti-inflammatory polarization, suggesting that YM, through an indirect mechanism, is able to skew the M1/M2 ratio. Moreover, YM consumption reduced the Eubacterium rectale/Clostridium coccoides and Enterobacteriaceae groups and increased the Lactobacillus/Lactococcus group in the gut microbiota. In summary, we show that YM promotes an immunosuppressive environment by enhancing anti-inflammatory M2 macrophage differentiation, reducing colitis symptoms, and suggesting that YM consumption may be a good cost-effective treatment for ulcerative colitis.

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Introdução: Investigações têm mostrado que a proteína anti-inflamatória Anexina A1 (ANXA1) está associada ao processo de invasão tumoral e que suas ações podem ser mediadas pelo receptor para peptídeos formilados (FPR), metaloproteinases (MMP-2 e MMP-9) e enzima de ciclooxigenase (COX-2). Por essas razões o objetivo do trabalho foi verificar a expressão da ANXA1 e dos receptores FPR1 e FPR2 em processos inflamatórios e tumorais do cólon. Material e Métodos: A detecção das proteínas foi realizada, por imuno-histoquímica, de biópsias de inflamação de cólon (colite) e tumores benignos (adenomas) e malignos (adenocarcinomas) de cólon, provenientes dos arquivos do Serviço de Patologia do Centro Universitário Padre Albino (UNIFIPA). As secções foram incubadas com os anticorpos primários policlonais anti-ANXA1 (1:1000), anti-FPR1, anti-FPR2 (1:500), anti-COX-2 (1:500), anti-MMP-2 (1:350) e anti-MMP-9 (1:350), reveladas com DAB e contracoradas com Hematoxilina. As quantificações foram feitas por densitometria e as análises estatísticas realizadas pela ANOVA e post test de Bonferroni. Resultados: Nossas análises imuno-histoquímicas e densitométricas mostraram aumento da expressão de ANXA1, FPR1 e COX-2 nos casos de colite, adenoma e adenocarcinoma, comparados a margem normal. Não houve imunorreatividade para FPR2 em nenhuma das condições clínicas estudadas. Também, foi observado aumento de MMP-2 nos casos de colite e adenocarcinoma e aumento de MM 9 no adenocarcinoma. Os dados mostram a correlação positiva da proteína ANXA1 com o receptor FPR1, COX-2 e MMP 2 nas biópsias de colite. Conclusões: A ANXA1 pode interagir de maneira específica com o receptor FPR1, COX-2 e MMP 2 nos processos inflamatórios e no desenvolvimento tumoral do cólon

Introduction: Investigations have shown that the anti-inflammatory protein Annexin A1 (ANXA1) is associated with the process of tumor invasion and that its actions can be mediated by the receptor for formylated peptides (FPR), metalloproteinases (MMP-2 and MMP-9) and cyclooxygenase enzyme (COX-2). For these reasons, the objective of the work was to verify the expression of ANXA1 and the FPR1 and FPR2 receptors in inflammatory and tumor processes of the colon. Material and Methods: Detection of proteins was carried out, by immunohistochemistry, from biopsies of colon inflammation (colitis) and benign (adenomas) and malignant (adenocarcinomas) colon tumors. The material was obtained from the archives of the Pathology Service of Padre Albino University Center (UNIFIPA). The sections were incubated with the primary polyclonal antibodies anti-ANXA1 (1:1000), anti-FPR1, anti-FPR2 (1:500), anti-COX-2 (1:500), anti-MMP-2 (1: 350) and anti-MMP-9 (1:350), revealed with DAB and counterstained with Hematoxylin. Quantifications were made by densitometry and statistical analyzes were carried out by ANOVA and Bonferroni post test. Results: Our immunohistochemical and densitometric analyzes showed increased expression of ANXA1, FPR1 and COX-2 in cases of colitis, adenoma and adenocarcinoma, compared to normal margins. There was no immunoreactivity for FPR2 in any of the clinical conditions studied. Also, an increase in MMP-2 was observed in cases of colitis and adenocarcinoma and an increase in MM-9 in adenocarcinoma. The data show the positive correlation of the ANXA1 protein with the FPR1, COX 2 and MMP-2 receptor in colitis biopsies. Conclusions: ANXA1 can specifically interact with the FPR1, COX-2 and MMP-2 receptor in inflammatory processes and colon tumor development

Introducción: Investigaciones han demostrado que la proteína antiinflamatoria Anexina A1 (ANXA1) está asociada al proceso de invasión tumoral y que sus acciones pueden ser mediadas por el receptor de péptidos formilados (FPR), metaloproteinasas (MMP-2 y MMP-9) y la enzima ciclooxigenasa (COX-2). Por estos motivos, el objetivo del trabajo fue verificar la expresión de ANXA1 y de los receptores FPR1 y FPR2 en procesos inflamatorios y tumorales del colon. Material y Métodos: La detección de proteínas se realizó, mediante inmunohistoquímica, a partir de biopsias de inflamación de colon (colitis) y tumores benignos (adenomas) y malignos (adenocarcinomas) de colon, del archivo del Servicio de Patología del Centro Universitario Padre Albino (UNIFIPA). Las secciones se incubaron con los anticuerpos policlonales primarios anti-ANXA1 (1:1000), anti-FPR1, anti-FPR2 (1:500), anti-COX-2 (1:500), anti-MMP-2 (1:350) y anti-MMP-9 (1:350), revelados con DAB y contrateñidos con hematoxilina. Las cuantificaciones se realizaron mediante densitometría y los análisis estadísticos se realizaron mediante ANOVA y post test de Bonferroni. Resultados: Nuestros análisis inmunohistoquímicos y densitométricos mostraron una mayor expresión de ANXA1, FPR1 y COX-2 en casos de colitis, adenoma y adenocarcinoma, en comparación con los márgenes normales. No hubo inmunorreactividad para FPR2 en ninguna de las condiciones clínicas estudiadas. Además, se observó un aumento de MMP-2 en casos de colitis y adenocarcinoma y un aumento de MM-9 en adenocarcinoma. Los datos muestran la correlación positiva de la proteína ANXA1 con los receptores FPR1, COX-2 y MMP-2 en biopsias de colitis. Conclusiones: ANXA1 puede interactuar específicamente con los receptores FPR1, COX-2 y MMP-2 en procesos inflamatorios y desarrollo de tumores de colon

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Lactobacillus delbrueckii subsp. lactis CIDCA 133 is a health-promoting bacterium that can alleviate gut inflammation and improve the epithelial barrier in a mouse model of mucositis. Despite these beneficial effects, the protective potential of this strain in other inflammation models, such as inflammatory bowel disease, remains unexplored. Herein, we examined for the first time the efficacy of Lactobacillus delbrueckii CIDCA 133 incorporated into a fermented milk formulation in the recovery of inflammation, epithelial damage, and restoration of gut microbiota in mice with dextran sulfate sodium-induced colitis. Oral administration of Lactobacillus delbrueckii CIDCA 133 fermented milk relieved colitis by decreasing levels of inflammatory factors (myeloperoxidase, N-acetyl-ß-D-glucosaminidase, toll-like receptor 2, nuclear factor-κB, interleukins 10 and 6, and tumor necrosis factor), secretory immunoglobulin A levels, and intestinal paracellular permeability. This immunobiotic also modulated the expression of tight junction proteins (zonulin and occludin) and the activation of short-chain fatty acids-related receptors (G-protein coupled receptors 43 and 109A). Colonic protection was effectively associated with acetate production and restoration of gut microbiota composition. Treatment with Lactobacillus delbrueckii CIDCA 133 fermented milk increased the abundance of Firmicutes members (Lactobacillus genus) while decreasing the abundance of Proteobacteria (Helicobacter genus) and Bacteroidetes members (Bacteroides genus). These promising outcomes influenced the mice's mucosal healing, colon length, body weight, and disease activity index, demonstrating that this immunobiotic could be explored as an alternative approach for managing inflammatory bowel disease.

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Introduction: Inflammatory bowel diseases (IBDs) disrupt the intestinal epithelium, leading to severe chronic inflammation. Current therapies cause adverse effects and are expensive, invasive, and ineffective for most patients. Annexin A1 (AnxA1) is a pivotal endogenous anti-inflammatory and tissue repair protein in IBD. Nanostructured compounds loading AnxA1 or its active N-terminal mimetic peptides improve IBD symptomatology. Methods: To further explore their potential as a therapeutic candidate, the AnxA1 N-terminal mimetic peptide Ac2-26 was incorporated into SBA-15 ordered mesoporous silica and covered with EL30D-55 to deliver it by oral treatment into the inflamed gut. Results: The systems SBA-Ac2-26 developed measurements revealed self-assembled rod-shaped particles, likely on the external surface of SBA-15, and 88% of peptide incorporation. SBA-15 carried the peptide Ac2-26 into cultured Raw 264.7 macrophages and Caco-2 epithelial cells. Moreover, oral administration of Eudragit-SBA-15-Ac2-26 (200 µg; once a day; for 4 days) reduced colitis clinical symptoms, inflammation, and improved epithelium recovery in mice under dextran-sodium sulfate-induced colitis. Discussion: The absorption of SBA-15 in gut epithelial cells is typically low; however, the permeable inflamed barrier can enable microparticles to cross, being phagocyted by macrophages. These findings suggest that Ac2-26 is successfully delivered and binds to its receptors in both epithelial and immune cells, aligning with the clinical results. Conclusion: Our findings demonstrate a simple and cost-effective approach to delivering Ac2-26 orally into the inflamed gut, highlighting its potential as non-invasive IBD therapy.

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The mammalian target of rapamycin (mTOR) pathway plays a key role in determining immune cells function through modulation of their metabolic status. By specific deletion of Rictor in CD11c+ myeloid cells (referred to here as CD11cRicΔ/Δ), we investigated the role of mTOR complex 2 (mTORC2) signaling in dendritic cells (DCs) function in mice. We showed that upon dextran sulfate sodium-induced colitis, the lack of mTORC2 signaling CD11c+ cells diminishes the colitis score and abrogates DC migration to the mesenteric lymph nodes, thereby diminishing the infiltration of T helper 17 cells in the lamina propria and subsequent inflammation. These findings corroborate with the abrogation of cytoskeleton organization and the decreased activation of Rac1 and Cdc42 GTPases observed in CD11c+-mTORC2-deficient cells. Meta-analysis on colonic samples from ulcerative colitis patients revealed increased gene expression of proinflammatory cytokines, which coincided with augmented expression of the mTOR pathway, a positive correlation between the DC marker ITGAX and interleukin-6, the expression of RICTOR, and CDC42. Together, this work proposes that targeting mTORC2 on DCs offers a key to hamper inflammatory responses, and this way, ameliorates the progression and severity of intestinal inflammatory diseases.

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Phytotherapy is an attractive strategy to treat inflammatory bowel disease (IBD) that could be especially useful in developing countries. We previously demonstrated the intestinal anti-inflammatory effect of the total ethereal extract from the Physalis peruviana (Cape gooseberry) calyces in TNBS-induced colitis. This work investigates the therapeutic potential of Peruviose A and B, two sucrose esters that constitute the major metabolites of its calyces. The effect of the Peruvioses A and B mixture on TNBS-induced colitis was studied after 3 (preventive) and 15-days (therapy set-up) of colitis induction in rats. Colonic inflammation was assessed by measuring macroscopic/histologic damage, MPO activity, and biochemical changes. Additionally, LPS-stimulated RAW 264.7 macrophages were treated with test compounds to determine the effect on cytokine imbalance in these cells. Peruvioses mixture ameliorated TNBS-induced colitis in acute (preventive) or established (therapeutic) settings. Although 3-day treatment with compounds did not produce a potent effect, it was sufficient to significantly reduce the extent/severity of tissue damage and the microscopic disturbances. Beneficial effects in the therapy set-up were substantially higher and involved the inhibition of pro-inflammatory enzymes (iNOS, COX-2), cytokines (TNF-α, IL-1ß, and IL-6), as well as epithelial regeneration with restoration of goblet cells numbers and expression of MUC-2 and TFF-3. Consistently, LPS-induced RAW 264.7 cells produced less NO, PGE2, TNF-α, IL-6, and MCP-1. These effects might be related to the inhibition of the NF-κB signaling pathway. Our results suggest that sucrose esters from P. peruviana calyces, non-edible waste from fruit production, might be useful as an alternative IBD treatment.

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Objective: to conduct a literature review on possible serum markers in patients with UC to identify potential biomarkers. Methods: the descriptors "Colitis Ulcerative", "Biomarkers", and "Diagnosis" were used for the search in the PUBMED, LILACS, SciELO, and SCOPUS databases. After applying the inclusion and exclusion criteria in the databases, the 32 articles were classified as samples. Results: research indicates that biomarkers can play distinct roles in assessing activity and diagnosing UC. For instance, the peptide nesfatin-1 showed elevated serum levels during active periods of the disease, while the peptide trefoil factor 3 showed promise in predicting UC activity. It is worth highlighting that C-reactive protein (CRP) is a commonly used marker in which high values are observed in patients during the active phase of the disease; however, studies suggest the need to redefine CRP cutoff values to better predict endoscopic remission. Other approaches, such as the analysis of proteins, specific fatty acids, and proteins related to the extracellular matrix, have also been explored, highlighting their potential as biomarkers. Conclusion: in summary, there is a diversity of possible clinically important biomarkers that play different roles in the assessment and diagnosis of UC. However, it is essential to conduct further research to validate these biomarkers and achieve greater reliability.

Objetivo: realizar uma revisão da literatura sobre possíveis marcadores séricos em pacientes com RU com o objetivo de identificar potenciais biomarcadores. Métodos: os descritores "Colitis Ulcerative", "Biomarkers" e "Diagnosis" foram utilizados para a busca nas bases de dados PUBMED, LILACS, SciELO e SCOPUS. Após a aplicação dos critérios de inclusão e exclusão nas bases de dados, os 32 artigos foram classificados como amostras. Resultados: a pesquisa indica que os biomarcadores podem desempenhar papéis distintos na avaliação da atividade e no diagnóstico da RU. Por exemplo, o peptídeo nesfatin-1 apresentou níveis séricos elevados durante os períodos ativos da doença, enquanto o peptídeo trefoil factor 3 mostrou-se promissor na predição da atividade da RU. Vale destacar que a proteína C-reativa (PCR) é um marcador comumente utilizado, no qual valores elevados são observados em pacientes durante a fase ativa da doença; no entanto, estudos sugerem a necessidade de redefinir os valores de corte da PCR para melhor predizer a remissão endoscópica. Outras abordagens, como a análise de proteínas, ácidos graxos específicos e proteínas relacionadas à matriz extracelular, também têm sido exploradas, destacando seu potencial como biomarcadores. Conclusão: em resumo, há uma diversidade de possíveis biomarcadores de importância clínica que desempenham diferentes papéis na avaliação e diagnóstico da CU. No entanto, é essencial conduzir mais pesquisas para validar esses biomarcadores para obter maior confiabilidade.

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A Anexina A1 (AnxA1) é uma proteína de 37 kDa que controla o desenvolvimento da reação inflamatória inata, e favorece a eferocitose e o reparo tecidual. Em doenças inflamatórias intestinais (DIIs), tanto a AnxA1 endógena, como a sintética e o peptídeo sintético mimético ao N-terminal da proteína (Ac2-26) inibem o desenvolvimento de doença e induzem a cicatrização. O presente projeto teve o objetivo de obter novas formulações para carrear a AnxA1 recombinante (rAnxA1) ou o Ac2-26 e testar suas eficácias no modelo de colite experimental induzida pelo dextram sulfato de sódio (DSS, 0-6 dias) em camundongos C57Bl/6 machos. A rAnxA1 foi funcionalizada em nanocápsulas de núcleo lipídico de parede múltipla (MLNC) pela ligação Zn2+, com alta eficência de incorporação (92%) e adminsitrada pelas vias oral, intravenosa ou intraperitoneal durante a fase latente da doença (6º-9º dia). Somente o tratamento intraperitoneal com MLNC-AnxA1 (12,5 µg/mL) reduziu significativamente os sinais clínicos da doença, restaurou a integridade da estrutura colônica e a proliferação celular, bem como aumentou expressão de junções celulares da barreira intestinal. Ainda, MLNC-AnxA1 induziu a polarização de macrófagos para o fenótipo M2 in vivo no tecido inflamado e in vitro após estímulo com lipopolissacarídeos (LPS) bacteriano. Na tentativa de obter uma formulação terapêutica com atividade por vial oral, o peptídeo Ac2-26 foi incorporado em sílica mesoporosa ordenada SBA-15 e revestidos com Eudragit® L30-D55. A incorporação do peptídeo foi efetiva (88%) e a administração oral de Eudragit-SBA15-Ac2-26 (6º-9º dia; 200 µg/camundongo; 8 mg/kg) reduziu significativamente os sintomas clínicos e inflamação. De fato, ensaios de PET-SCAN mostraram que o SBA-15 permaneceu no intestino por até 16 horas após a administração e promoveu a liberação do peptídeo no intestino inflamado. Em cultura celular de epitélio (Caco-2), Eudragit-SBA15-Ac2-26 favoreceu a internalização de Ac2-26. Em conjunto, as duas estratégias expermentais de entrega do rAnxA1 ou Ac2-26 foram eficientes e os resultados obtidos sugerem que mais estudos devem ser realizados para a confirmação das estratégias de tratamento. Com o intuito de buscar ferramentas para ampliar estes estudos, durante estágio BEPE foram realizados estudos em cultura de células epiteliais baseado em células-tronco adultas diferenciadas in vitro. Os resultados mostraram que rAnxA1 ou Ac2-26 protegeram a integridade epitelial após desafio com LPS, pela regulação positiva da expressão das junções oclusivas e aderentes e redução da expressão de claudina-2, responsável pelo aumento da permeabilidade intercelular; pela modulação negativa decitocinas pró-inflamatórias CXCL-1 e MCP-1, e positiva de citocina antiinflamatória IL-10. Desta forma, padronizamos um novo modelo de cultura celular ainda não testada para a AnxA1 ou Ac2-26, que poderá ser empregada para desvendar os mecanismos da MLNC-AnxA1 e do Eudragit-SBA15-Ac2-26

Annexin Al (AnxA1) is a 37 kDa protein that controls the development of the innate inflammatory reaction, and favors efferocytosis and tissue repair. In inflammatory bowel diseases (IBDs), both endogenous and synthetic AnxA1 and the synthetic peptide mimetic to the N-terminal of the protein (Ac2-26) inhibit the development of disease and induce healing. The present project aimed to obtain new formulations to carry recombinant AnxA1 (rAnxA1) or Ac2-26 and test their efficacy in the experimental colitis model induced by dextram sodium sulfate (DSS, 0-6 days) in C57Bl/6 mice. rAnxA1 was functionalized into multiwall lipid core nanocapsules (MLNC) by Zn2+ binding, with high incorporation efficiency (92%) and administered orally, intravenously or intraperitoneally during the latent phase of the disease (6º-9º day). Only intraperitoneal treatment with MLNC-AnxA1 (12.5 µg/mL) significantly reduced the clinical signs of the disease, restored the integrity of the colonic structure and cell proliferation, as well as increased the expression of intestinal barrier cell junctions. Furthermore, MLNC-AnxA1 induced macrophage polarization to the M2 phenotype in vivo in inflamed tissue and in vitro after stimulation with bacterial lipopolysaccharides (LPS). In an attempt to obtain a therapeutic formulation with oral activity, the Ac2-26 peptide was incorporated into ordered mesoporous silica SBA-15 and coated with Eudragit® L30-D55. Peptide incorporation was effective (88%) and oral administration of Eudragit-SBA15-Ac2-26 (6º-9º day; 200 µg/mice; 8 mg/kg) significantly reduced clinical symptoms and inflammation. PET-SCAN assays showed SBA-15 remained in the intestine for up to 16 hours after administration and promoted the release of the peptide in the inflamed intestine. In epithelial cell culture (Caco-2), SBA15-Ac2-26 favored the internalization of Ac2-26. Taken together, the two experimental delivery strategies for rAnxA1 or Ac2-26 were efficient and the results obtained suggest that more studies should be carried out to confirm the treatment strategies. In order to seek tools to expand these studies, during the BEPE internship, studies were carried out in epithelial cell cultures based on adult stem cells differentiated in vitro. The results showed rAnxA1 or Ac2-26 protected epithelial integrity after challenge with LPS, by upregulating the expression of tight and adherens junctions and reducing the expression of claudin-2, responsible for increasing intercellular permeability; by negative modulation of pro-inflammatory cytokines CXCL-1 and MCP-1, and positive modulation of anti-inflammatory cytokine IL-10. In this way, we standardized a new cell culture model that has not yet been tested for AnxA1 or Ac2-26, which could be used to unravel the mechanisms of MLNC-AnxA1 and Eudragit-SBA15-Ac2-26

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Eosinophilic colitis is a rare and severe form of eosinophilic gastrointestinal disease. This study presents the case of a 69-year-old woman from Anadia, Alagoas, Brazil, who experienced intermittent abdominal pain in the right and left iliac fossae, along with recurrent eructations, flatus, in-creased bowel movements, diarrhea, and loss of appetite for approximately six months. The diagnosis of eosinophilic colitis was confirmed through upper digestive endoscopy, colonoscopy, and histopathology, which revealed eosinophilic pancolitis with about 100 eosinophils per high-power field in most tissue fragments. After three months of a restricted diet and treatment with ketotifen 1 mg (twice daily for 30 days) and esomeprazole 40 mg (once daily on an empty stomach for 56 days), the patient showed clinical improvement. This case highlights the diagnostic challenges of eosinophilic colitis due to its non-specific clinical signs and emphasizes the necessity of supplementary examinations (AU)

A colite eosinofílica é uma doença rara, sendo a forma menos frequente e mais grave entre as doenças eosinofílicas gastrintestinais. Este estudo apresenta o caso de uma mulher de 69 anos, natural de Anadia, Alagoas, Brasil, que apresentou dor abdominal intermitente nas fossas ilíacas direita e esquerda, acompanhada de eructações, flatos, aumento da frequência do trânsito intestinal, diarreia e inapetência por aproximadamente seis meses. O diagnóstico de colite eosinofílica foi confirmado por endoscopia digestiva alta, colonoscopia e histopatologia, que revelaram pancolite eosinofílica com cerca de 100 eosinófilos por campo de grande aumento na maioria dos fragmentos de tecido. Após três meses de dieta restritiva e tratamento com cetotifeno 1 mg (duas vezes ao dia por 30 dias) e esomeprazol 40 mg (uma vez ao dia em jejum por 56 dias), a paciente apresentou melhora clínica. Este caso destaca os desafios diagnósticos da colite eosinofílica devido aos seus sinais clínicos inespecíficos e enfatiza a necessidade de exames complementares (AU)

RESUMO

Eosinophilic colitis (EoC), a rare immune-mediated disease that is part of the eosinophilic gastrointestinal diseases, is characterized by the presence of an eosinophilic infiltrate in the colonic wall in symptomatic patients. Before considering the diagnosis of EoC, other diseases associated with colonic eosinophilia should be ruled out, such as parasitic infections, drugs, chronic immune-mediated diseases, and neoplasms. The symptoms of EoC are variable and non-specific, being abdominal pain and diarrhea the most common. Although systemic corticosteroids and budesonide have demonstrated their efficacy, these drugs have only been evaluated in case series studies and cli- nical case reports. Herein, we discuss the clinical strategy for diagnosis, therapy selection, and follow-up of EoC

La colitis eosinofílica (CEo), una enfermedad inmunomediada que forma parte de las enfermedades gastrointes- tinales eosinofílicas, se caracteriza por la presencia de infiltrado eosinofílico en la pared del colon en pacientes sintomáticos. Antes de plantear el diagnóstico de una CEo, otras enfermedades asociadas a una eosinofilia coló- nica, incluyendo infecciones parasitarias, fármacos, enfermedades crónicas inmunomediadas y neoplasias, deben ser descartadas. Los síntomas de la CEo son variables e inespecíficos, siendo el dolor abdominal y la diarrea los más frecuentes. Aunque los corticoides sistémicos y la budesonida han demostrado su eficacia, estos fármacos han sido evaluados solo en estudios de serie de casos y reportes de casos clínicos. En este artículo, discutimos la estrategia clínica para el diagnóstico, selección del tratamiento y el seguimiento de la CEo.

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Ulcerative colitis (UC) is an important chronic and multifactorial disease, which alters the colon mucosal with a significant impact on life quality affecting both men and women. The difference between genders causes changes in the inflammatory processes, modulating the development of several diseases. The available drugs to treat UC exhibit limited outcomes and side effects; thus, new therapies are needed. Photobiomodulation (PBM) emerges as potential treatment by modulating the inflammatory process without side effects and low costs. The aim of this study was to evaluate the effects of PBM in acetic acid-induced UC comparing the responses between male and females. For this purpose, male and female Wistar rats (36) were submitted to induction of UC by rectal administration of 10% acetic acid (colitis group) and treated or not with PBM (colitis-PBM group) (LED, 660 nm, 100 mW, 150 s) in three points: right side and left of the ventral surface and in the external anal region. Non-manipulated rats were used as control (basal group). We investigated the disease activity index (DAI score), myeloperoxidase enzyme activity (MPO) and release of cytokines in the intestine homogenates, and histological analysis. PBM reduces DAI score, MPO activity, and mast cell degranulation while increased mucous production in both females and males. Moreover, PBM reduced histopathological score as well as the levels of IL-6 and IL-4 in the bowel only in males. We also showed reduced levels of IL-1beta and TNF-alpha after PBM in both males and females, while the levels of IL-10 and IFN-gamma were increased. In conclusion, despite our study has shown some differences between males and females, PBM attenuated the biomarkers of UC in both genders constituting a potential combined treatment that is non-invasive and low cost.

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INTRODUCTION: Treatments of Inflammatory Bowel Disease (IBD) are able to control symptoms in most cases, however, a fraction of patients do not improve or have a loss of response to treatments, making it important to explore new therapeutic strategies. Hyperbaric oxygen therapy (HBO) may represent one of them. The aim of this study was to evaluate the effects of HBO therapy in an experimental model of IBD. METHODS: Sixty male BALBc mice were divided into six groups. Group 1 was colitis-induced with trinitrobenzene sulfonic acid (TNBS) + ethanol, group 2 received TNBS + ethanol plus HBO, group 3 received only ethanol, group 4 received ethanol plus HBO, group 5 received saline solution, and group 6 received saline solution plus HBO. HBO was performed for four days, subsequently, the mice were evaluated daily. At the end of the study, samples from the intestine were collected for histological analysis as well as for measurement of antioxidant enzymes and cytokine levels. RESULTS: HBO significantly improved the clinical and histological status of the animals. Treatment with HBO increased the activity of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) in all of the groups; moreover, the difference was only significant between the TNBS and TNBS + HBO groups and treatments promoted a reduction in the proinflammatory cytokines IFN-γ, IL-12, IL-17 and TNF-α and increased the anti-inflammatory cytokines IL-4 and IL-10, with no changes in IL-13. CONCLUSION: HBO effectively treats TNBS-induced colitis by increasing the activity of antioxidant enzymes and modulating cytokine profiles.

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AIMS: The carotid bodies are sensors that detect physiological signals and convey them to the central nervous system, where the stimuli are processed inducing reflexes through efferent pathways. Recent studies have demonstrated that electrical stimulation of the carotid sinus nerve (CSN) triggers the anti-inflammatory reflex under different conditions. However, whether this electrical stimulation attenuates colitis was never examined. This study aimed to evaluate if the electrical CSN stimulation attenuates the experimental colitis induced by intrarectal administration of acetic acid in rats. METHODS: Electrodes were implanted around the CSN to stimulate the CSN, and a catheter was inserted into the left femoral artery to record the arterial pressure. The observation of hypotensive responses confirmed the effectiveness of the electrical CNS stimulation. This maneuver was followed by a 4 % acetic acid or saline administered intrarectally. After 24 h, colons were segmented into distal and proximal parts for macroscopy, histological and biochemical assessment. KEY FINDINGS: As expected, the electrical CSN stimulation was effective in decreasing arterial pressure in saline and colitis rats. Moreover, electrical CSN stimulation effectively reduced colonic tissue lesions, colitis scores, and histopathologic parameters associated with colitis. In addition, the CSN stimulation also reduced the colonic mucosa pro-inflammatory cytokine interleukin-1 beta, and increased the anti-inflammatory interleukin-10, in rats submitted to colitis. SIGNIFICANCE: These findings indicated that electrical CSN stimulation breaks the vicious cycle of local colon inflammation in colitis, which might contribute to its better outcome.

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The role of cyclooxygenase (COXs) isoforms in maintaining colonic mucosal integrity is not fully understood. This study aimed to evaluate the role of COX-1 and -2 on colonic mucosal integrity in an experimental colitis model. Colitis was induced in Wistar rats by intracolonic administration of 2,4,6-trinitrobenzenesulfonic acid (20 mg + 50% ethanol). The control group (sham group) received saline only. After 7, 14, or 28 days, colonic samples were removed, and macroscopic lesion scores, wet weight, myeloperoxidase activity, and transepithelial electrical resistance (TER) were determined. In other rat groups, colonic samples from the sham group and a 7th day post-colitis group were mounted in Üssing chambers with the luminal side exposed to a buffer solution (control), acetylsalicylic acid (ASA), SC-560 (COX-1 inhibitor), or celecoxib (COX-2 inhibitor). TER and epithelial permeability to fluorescein were measured. The 7th day colitis group had higher macroscopic damage scores, wet weight, and myeloperoxidase activity and lower basal TER than the sham, 14th day colitis, and 28th day colitis groups. Inhibition of COX-1 but not COX-2 significantly decreased TER and increased permeability to fluorescein in the 7th day post-colitis group compared to the sham group. Additionally, ASA decreased the colonic mucosal integrity on day seven post-colitis compared to the sham group. A decrease in the colonic mucosa integrity in the experimental colitis model can be aggravated only by the inhibition of COX-1, which demonstrated the importance of this enzyme in the maintenance of colonic mucosal integrity.

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PURPOSE: To evaluate the tissue content of neutral and acidic mucins, sulfomucins and sialomucins in colonic glands devoid of intestinal transit after enemas containing sucralfate and n-acetylcysteine alone or in combination. METHODS: Sixty-four rats underwent intestinal transit bypass. A colonic segment was collected to compose the white group (without intervention). After derivation, the animals were divided into two groups according to whether enemas were performed daily for two or four weeks. Each group was subdivided into four subgroups according to the substance used: control group: saline 0.9%; sucralfate group (SCF): SCF 2 g/kg/day; n-acetylcysteine group (NAC): NAC 100 mg/kg/day; and SCF+NAC group: SCF 2 g/kg/day + NAC 100 mg/kg/day.Neutral and acidic mucins were stained by periodic acid-Schiff and alcian-blue techniques, respectively. The distinction between sulfomucins and sialomucin was made by the high alcian-blue iron diamine technique. The content of mucins in the colonic glands was measured by computerized morphometry. The inflammatory score was assessed using a validated scale. The results between the groups were compared by the Mann-Whitney's test, while the variation according to time by the Kruskal-Wallis' test (Dunn's post-test). A significance level of 5% was adopted. RESULTS: There was reduction in the inflammatory score regardless of the application of isolated or associated substances. Intervention with SCF+NAC increased the content of all mucin subtypes regardless of intervention time. CONCLUSIONS: The application of SCF+NAC reduced the inflammatory process of the colonic mucosa and increased the content of different types of mucins in the colonic glands of segments excluded from fecal transit.

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The management of inflammatory bowel diseases has been widely investigated, especially ulcerative colitis. Thus, studies with the application of new probiotic products are needed in the prevention/treatment of these clinical conditions. The objective of this work was to evaluate the effects of probiotic orange juice containing Pediococcus acidilactici CE51 in a murine model of colitis. 45 male Swiss lineage mice were used, divided into five groups (n = 9): control, colitis, colitis + probiotic (probiotic orange juice containing CE51), colitis + placebo (orange juice) and colitis + sulfasalazine (10 mg/kg/Weight). The induction of colitis was performed with dextran sodium sulfate (3%). The treatment time was 5 and 15 days after induction. Histopathological analysis, serum measurements of TNF-α and C-reactive protein and metagenomic analysis of feces were performed after euthanasia. Probiotic treatment reduced inflammation in the small intestine, large intestine and spleen. The probiotic did not alter the serum dosages of TNF-α and C-reactive protein. Their use maintained the quantitative ratio of the phylum Firmicutes/Bacteroidetes and increased Lactobacillus helveticus with 15 days of treatment (p < 0.05). The probiotic orange juice containing P. acidilactici CE51 positively modulated the gut microbiota composition and attenuated the inflammation induced in colitis.