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1.
J Agric Food Chem ; 67(49): 13758-13766, 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31789514

RESUMO

Probiotics, such as Lactobacillus, have been proven to be effective in maintaining intestinal homeostasis. The modulatory effect of Lactobacillus on intestinal epithelial development in early life is still unclear. In this study, Lactobacillus isolates with good probiotic abilities were screened and orally administered to detect their regulatory effect on intestinal development in chickens. L. reuteri 22 was isolated from chickens and chosen for subsequent chicken experiments due to its strong acid and bile salt resistance and ability to adhere to epithelial cells. The 3-day-old chickens were orally administrated with 108 CFU L. reuteri 22 for consecutive 7 days. L. reuteri 22 increased Lgr5 mRNA expression (3.23 ± 0.40, P = 0.001) and activated the Wnt/ß-catenin signaling pathway, with increasing expression of proliferating cell nuclear antigen (PCNA) (49.27 ± 9.81, P = 0.021) to support the proliferation of chicken intestinal epithelial cells. Moreover, L. reuteri 22 also inhibited the Notch signaling pathway to induce intestinal stem cell differentiation into goblet cells with increased mucin 2 (Muc-2) expression (1.72 ± 0.34, P = 0.047). L. reuteri 22 significantly enhanced lysozyme mRNA expression (2.32 ± 0.55, P = 0.019) to improve intestinal innate mucosal immunity. This study demonstrated that L. reuteri administration could regulate chicken intestinal epithelium development to ensure the function of the intestinal mucosal barrier, which is beneficial for newborn animals.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Epiteliais/citologia , Células Caliciformes/efeitos dos fármacos , Lactobacillus reuteri/fisiologia , Probióticos/farmacologia , Animais , Galinhas , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Caliciformes/citologia , Células Caliciformes/metabolismo , Intestinos/citologia , Intestinos/efeitos dos fármacos , Mucina-2/genética , Mucina-2/metabolismo , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Transdução de Sinais/efeitos dos fármacos , beta Catenina/genética , beta Catenina/metabolismo
2.
J Med Food ; 22(10): 1009-1021, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31536448

RESUMO

Constipation is a common gastrointestinal disorder characterized by changes in intestinal habits. Increasing evidence indicates that long-term use of irritant laxatives causes serious side effects. Meanwhile, more than 50% of patients are dissatisfied with sense of use of non-prescriptional laxatives. ß-glucans are natural polysaccharides widely found in yeast, fungus, and plants, which have been reported to exhibit various pharmacological effects. The aim of this study was to characterize the effect of ß-glucans extracted from the bread yeast cell wall on loperamide-induced constipation mice. Forty mice were fed with loperamide (10 mg/kg) to make the constipation model and a diet supplemented with 2.5, 5, and 10 mg/kg ß-glucan. We assessed the defecation frequency, intestinal transit function of mice, as well as used high-throughput sequencing to analyze the intestinal microbiota composition and functional biological profiles data. Meanwhile, we detected expression of neurotransmitters including acetylcholinesterase, substance P, and serotonin (5-HT) and expression of tight junction protein (TJP) including zonula occludens-1 and mucin-2 in distal colon to characterize the possible molecular mechanisms. ß-glucans significantly enhanced intestinal motility and provided a possibility to regulate the expression of neurotransmitters and TJP in mice. The intestinal microecological portion of the treatment group partially recovered and was closer to the normal group. This study showed that ß-glucans can influence the intestinal microbiota and restore microecological balance to regulate the express of neurotransmitters and TJP to recover intestinal epithelial mechanical barrier. We suggested that ß-glucans could be used as an active nutritional supplement to protect the damaged intestinal barrier and help patients who have constipation complications and dysbiosis.


Assuntos
Constipação Intestinal/tratamento farmacológico , Microbioma Gastrointestinal , Loperamida/efeitos adversos , Saccharomyces cerevisiae/química , beta-Glucanas/farmacologia , Acetilcolinesterase/metabolismo , Animais , Constipação Intestinal/induzido quimicamente , Defecação/efeitos dos fármacos , Trânsito Gastrointestinal/efeitos dos fármacos , Masculino , Metagenoma , Camundongos , Camundongos Endogâmicos BALB C , Mucina-2/metabolismo , Serotonina/metabolismo , Substância P/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
3.
Infect Immun ; 87(12)2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31527129

RESUMO

Epidemiological studies suggest frequent association of enteropathogenic bacteria with Entamoeba histolytica during symptomatic infection. In this study, we sought to determine if the interaction with enteropathogenic (EPEC) or nonpathogenic Escherichia coli (strain DH5α) could modify the virulence of E. histolytica to cause disease in animal models of amebiasis. In vitro studies showed a 2-fold increase in CaCo2 monolayer destruction when E. histolytica interacted with EPEC but not with E. coli DH5α for 2.5 h. This was associated with increased E. histolytica proteolytic activity as revealed by zymogram analysis and degradation of the E. histolytica CP-A1/5 (EhCP-A1/5) peptide substrate Z-Arg-Arg-pNC and EhCP4 substrate Z-Val-Val-Arg-AMC. Additionally, E. histolytica-EPEC interaction increased EhCP-A1, -A2, -A4, and -A5, Hgl, Apa, and Cox-1 mRNA expression. Despite the marked upregulation of E. histolytica virulence factors, nonsignificant macroscopic differences in amebic liver abscess development were observed at early stages in hamsters inoculated with either E. histolytica-EPEC or E. histolytica-E. coli DH5α. Histopathology of livers of E. histolytica-EPEC-inoculated animals revealed foci of acute inflammation 3 h postinoculation that progressively increased, producing large inflammatory reactions, ischemia, and necrosis with high expression of il-1ß, ifn-γ, and tnf-α proinflammatory cytokine genes compared with that in livers of E. histolytica-E. coli DH5α-inoculated animals. In closed colonic loops from mice, intense inflammation was observed with E. histolytica-EPEC manifested by downregulation of Math1 mRNA with a corresponding increase in the expression of Muc2 mucin and proinflammatory cytokine genes il-6, il-12, and mcp-1 These results demonstrate that E. histolytica/EPEC interaction enhanced the expression and production of key molecules associated with E. histolytica virulence, critical in pathogenesis and progression of disease.


Assuntos
Entamoeba histolytica/patogenicidade , Entamebíase/patologia , Escherichia coli Enteropatogênica/fisiologia , Interações entre Hospedeiro e Microrganismos/fisiologia , Animais , Células CACO-2 , Linhagem Celular , Cricetinae , Cisteína Proteases/metabolismo , Citocinas/metabolismo , Entamoeba histolytica/microbiologia , Células HT29 , Humanos , Inflamação , Mesocricetus , Camundongos , Camundongos Endogâmicos C57BL , Mucina-2/metabolismo , Fatores de Virulência/biossíntese
4.
J Biochem Mol Toxicol ; 33(11): e22397, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31557363

RESUMO

Fumonisins (Fums) are mycotoxins widely distributed in crops and feed, and ingestion of Fums-contaminated crops is harmful to animal health. The purpose of this study is to explore the effect of Fum B1 (FB1 ) on barrier functions of porcine intestinal epithelial cells, IPEC-J2, to clarify the intestinal toxicity of Fums in pigs. The results showed that the persistent treatment of FB1 significantly decreased the viability of IPEC-J2. Moreover, the expressions of Claudin 1, Occludin, Zonula Occluden-1 (ZO-1) on the messenger RNA (mRNA), and protein levels and MUC1 on the mRNA level were significantly inhibited after FB1 treatment, while the mRNA relative expression level of MUC2 was clearly increased. FB1 also enhanced the monolayer cell permeability of IPEC-J2. Importantly, FB1 promoted the expression of phosphorylated extracellular regulated protein kinase (p-ERK1/2 ). These data suggest that long-term treatment of FB1 can suppress IPEC-J2 proliferation, damage tight junctions of IPEC-J2, and regulate expression of mucins to induce the damage of barrier functions of porcine intestinal epithelial cells, which may be associated with the ERK1/2 phosphorylation pathway.


Assuntos
Células Epiteliais/metabolismo , Fumonisinas/farmacologia , Mucosa Intestinal/citologia , Micotoxinas/farmacologia , Permeabilidade/efeitos dos fármacos , Junções Íntimas/efeitos dos fármacos , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Fusarium/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Mucina-1/genética , Mucina-1/metabolismo , Mucina-2/genética , Mucina-2/metabolismo , Fosforilação/efeitos dos fármacos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacos , Suínos , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/metabolismo
5.
Nat Commun ; 10(1): 4003, 2019 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-31488830

RESUMO

Members of the interleukin-1 (IL-1) family are important mediators of obesity and metabolic disease and have been described to often play opposing roles. Here we report that the interleukin-36 (IL-36) subfamily can play a protective role against the development of disease. Elevated IL-36 cytokine expression is found in the serum of obese patients and negatively correlates with blood glucose levels among those presenting with type 2 diabetes. Mice lacking IL-36Ra, an IL-36 family signalling antagonist, develop less diet-induced weight gain, hyperglycemia and insulin resistance. These protective effects correlate with increased abundance of the metabolically protective bacteria Akkermansia muciniphila in the intestinal microbiome. IL-36 cytokines promote its outgrowth as well as increased colonic mucus secretion. These findings identify a protective role for IL-36 cytokines in obesity and metabolic disease, adding to the current understanding of the role the broader IL-1 family plays in regulating disease pathogenesis.


Assuntos
Citocinas/metabolismo , Microbioma Gastrointestinal/fisiologia , Interleucina-1/metabolismo , Doenças Metabólicas/metabolismo , Obesidade/metabolismo , Animais , Colo/imunologia , Colo/microbiologia , Colo/patologia , Diabetes Mellitus Tipo 2 , Microbioma Gastrointestinal/imunologia , Expressão Gênica , Teste de Tolerância a Glucose , Interações entre Hospedeiro e Microrganismos/imunologia , Interações entre Hospedeiro e Microrganismos/fisiologia , Humanos , Mediadores da Inflamação/metabolismo , Resistência à Insulina , Interleucina-1/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mucina-2/metabolismo , Obesidade/imunologia , Receptores de Interleucina-1/genética , Receptores de Interleucina-1/metabolismo , Transcriptoma , Verrucomicrobia
6.
Nat Commun ; 10(1): 4306, 2019 09 20.
Artigo em Inglês | MEDLINE | ID: mdl-31541089

RESUMO

The mucus layer is the first line of innate host defense in the gut that protects the epithelium by spatially separating commensal bacteria. MUC2 mucin is produced and stored by goblet cells that is constitutively exocytosed or hyper secreted upon sensing a threat. How coordinated mucus exocytosis maintains homeostasis in the intestinal epithelium and modulates the immunological landscape remains elusive. Here we describe how the vesicle SNARE protein VAMP8 coordinates mucin exocytosis from goblet cells. Vamp8-/- exhibit a mild pro-inflammatory state basally due to an altered mucus layer and increased encounters with microbial antigens. Microbial diversity shifts to a detrimental microbiota with an increase abundance of pathogenic and mucolytic bacteria. To alleviate the heavy microbial burden and inflammatory state basally, Vamp8-/- skews towards tolerance. Despite this, Vamp8-/- is highly susceptible to both chemical and infectious colitis demonstrating the fragility of the intestinal mucosa without proper mucus exocytosis mechanisms.


Assuntos
Colo/metabolismo , Exocitose/fisiologia , Células Caliciformes/metabolismo , Mucosa Intestinal/metabolismo , Mucina-2/metabolismo , Proteínas R-SNARE/metabolismo , Animais , Biodiversidade , Colo/patologia , Citocinas/metabolismo , Células Caliciformes/patologia , Homeostase , Humanos , Intestinos/patologia , Camundongos Knockout , Microbiota , Mucina-2/genética , Muco/metabolismo , Fenótipo , Proteínas R-SNARE/genética , Proteínas SNARE/metabolismo
7.
Nutrients ; 11(9)2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31514316

RESUMO

Diets high in saturated fatty acids (FA) represent a risk factor for the development of obesity and associated metabolic disorders, partly through their impact on the epithelial cell barrier integrity. We hypothesized that unsaturated FA could alleviate saturated FA-induced endoplasmic reticulum (ER) stress occurring in intestinal secretory goblet cells, and consequently the reduced synthesis and secretion of mucins that form the protective mucus barrier. To investigate this hypothesis, we treated well-differentiated human colonic LS174T goblet cells with palmitic acid (PAL)-the most commonly used inducer of lipotoxicity in in vitro systems-or n-9, n-6, or n-3 unsaturated fatty acids alone or in co-treatment with PAL, and measured the impact of such treatments on ER stress and Muc2 production. Our results showed that only eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids protect goblet cells against ER stress-mediated altered Muc2 secretion induced by PAL, whereas neither linolenic acid nor n-9 and n-6 FA are able to provide such protection. We conclude that EPA and DHA could represent potential therapeutic nutrients against the detrimental lipotoxicity of saturated fatty acids, associated with type 2 diabetes and obesity or inflammatory bowel disease. These in vitro data remain to be explored in vivo in a context of dietary obesity.


Assuntos
Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Células Caliciformes/efeitos dos fármacos , Mucina-2/metabolismo , Ácido Palmítico/toxicidade , Linhagem Celular , Citoproteção , Células Caliciformes/metabolismo , Células Caliciformes/patologia , Humanos , Fatores de Transcrição Kruppel-Like/metabolismo , Via Secretória
8.
Environ Pollut ; 254(Pt A): 112960, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31394344

RESUMO

That an alteration of the intestinal permeability is associated with gut barrier function has been increasingly evident, which plays an important role in human and animal health. Bisphenol A (BPA), an industrial compound used worldwide, has recently been classified as an environmental pollutant. One of our earlier studies has demonstrated that BPA disrupts the intestinal barrier function by inducing apoptosis and inhibiting cell proliferation in the human colonic epithelial cells line. In this study, we investigated the effects of dietary BPA uptake on the colonic barrier function in mice, as well as the intestinal permeability. Dietary BPA uptake was observed to destroy the morphology of the colonic epithelium and increase the pathology score. The levels of endotoxin, diamine peroxidase, D-lactate, and zonulin were found to have been significantly elevated in both plasma and colonic mucosa. A decline in the number of intestinal goblet cells and in mucin 2 gene expression was observed in the mice belonging to the BPA group. The results of immunohistochemistry revealed that the expression of tight junction proteins (ZO-1, occludin, and claudin-1) in colonic epithelium of BPA mice decreased significantly, and their gene abundance was also inhibited. Moreover, dietary BPA uptake was also found to have significantly reduced colonic microbial diversity and altered microbial structural composition. The functional profiles of colonic bacterial community exhibited adverse effects of dietary BPA intake on the endocrine and digestive systems, as well as the transport and catabolism functions. Collectively, our study highlighted that dietary BPA increased the colonic permeability, and this effect was closely related to the disruption of intestinal chemistry and physical and biological barrier functions.


Assuntos
Compostos Benzidrílicos/toxicidade , Substâncias Perigosas/toxicidade , Intestinos/efeitos dos fármacos , Fenóis/toxicidade , Animais , Apoptose/efeitos dos fármacos , Colo , Humanos , Mucosa Intestinal/metabolismo , Masculino , Camundongos , Mucina-2 , Ocludina , Permeabilidade/efeitos dos fármacos
9.
J Agric Food Chem ; 67(35): 9831-9839, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31407897

RESUMO

Probiotic lactobacilli and their exopolysaccharides (EPS) are thought to modulate mucosal homeostasis; however, their mechanisms remain elusive. Thus, we tried to clarify the role of exopolysaccharides from Lactobacillus plantarum NCU116 (EPS116) in the intestinal mucosal homeostasis. Our results indicated that EPS116 regulated the colon mucosal healing and homeostasis, enhanced the goblet cell differentiation, and promoted the expression of Muc2 gene in vivo and in vitro. Further experiments showed that EPS116 promoted the expression and phosphorylation of transcription factor c-Jun and facilitated its binding to the promoter of Muc2. Moreover, knocking down c-Jun or inhibiting its function in LS 174T cells treated with EPS116 led to decreased expression of Muc2, implying that EPS116 promoted the colonic mucosal homeostasis and Muc2 expression via c-Jun. Therefore, our study uncovered a novel model where EPS116 enhanced colon mucosal homeostasis by controlling the epithelial cell differentiation and c-Jun/Muc2 signaling.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Colo/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Lactobacillus plantarum/química , Mucina-2/metabolismo , Polissacarídeos Bacterianos/farmacologia , Proteínas Proto-Oncogênicas c-jun/metabolismo , Animais , Linhagem Celular Tumoral , Colo/citologia , Colo/metabolismo , Colo/fisiopatologia , Homeostase/efeitos dos fármacos , Humanos , Mucosa Intestinal/citologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucina-2/genética , Regiões Promotoras Genéticas , Proteínas Proto-Oncogênicas c-jun/genética , Transdução de Sinais/efeitos dos fármacos
10.
Acta Clin Croat ; 58(1): 23-28, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31363321

RESUMO

Higher expression of the mucin 2 (MUC2) glycoprotein and vascular endothelial growth factor (VEGF) in Barrett's mucosa may be associated with a higher risk of esophageal adenocarcinoma development. Thirty-six patients diagnosed with Barrett's esophagus (BE), short-segment, were included in the study due to unsuccessful treatment with proton pump inhibitors. The diagnosis was confirmed by histopathologic analysis of the tissue obtained by esophagogastric junction biopsy. Expression of MUC2 and VEGF was determined by immunohistochemistry. We found four patients in early stage of adenocarcinoma and 32 patients with BE; five of them had indication for argon plasma coagulation treatment, one for radiofrequency ablation and one for endoscopic mucosal resection; 25 patients were treated with proton pump inhibitors. Regression of BE occurred in 25 (69.44%) patients. MUC2 positivity is unique for goblet cells in patients with BE, but it is not the only marker. VEGF is an indicator of angiogenesis in the mucosa of patients with BE and adenocarcinoma.


Assuntos
Adenocarcinoma/metabolismo , Esôfago de Barrett/metabolismo , Neoplasias Esofágicas/metabolismo , Esôfago/metabolismo , Mucina-2/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adenocarcinoma/patologia , Idoso , Esôfago de Barrett/patologia , Biópsia , Transformação Celular Neoplásica , Neoplasias Esofágicas/patologia , Esôfago/patologia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Membrana Mucosa/patologia
11.
Food Funct ; 10(7): 4315-4329, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31271400

RESUMO

In this study, the immunostimulatory activity of Caulerpa lentillifera polysaccharides (CLP) was elucidated in cytoxan (CTX)-induced immunosuppressed BALB/c mice. The results showed that CLP ameliorated the CTX-evoked damage to body weight, colon length and thymus/spleen indexes and enhanced the secretions of interleukin (IL)-1ß, tumor necrosis factor (TNF)-α and superoxidase dismutase (SOD) in serum and thymic, splenic and colonic tissues of the immunosuppressed mice. Besides, CLP promoted the production of secretory immunoglobulin A (SIgA) and mucin2 in the colonic tissue of the immunosuppressed mice. Associated with the above immunostimulatory effects, CLP positively affected the production of short chain fatty acids (SCFAs) and microbiota diversity and composition, such as improvement in the growth of Lactobacillus, Coriobacteriaceae, Ruminococcaceae, Clostridium_XVIII and Helicobacter, whereas it suppressed the microbial populations of Bacteroides, Barnesiella and Lachnospiraceae. These findings suggested that CLP modulated SCFA production and gut microbiota in the immunosuppressed mice, evoking the colonic mucosal immunity, which might activate the systemic immunity in blood, thymus and spleen. The results could be helpful for understanding the functions of CLP, supporting their potential as novel prebiotics and immunostimulators.


Assuntos
Adjuvantes Imunológicos/farmacologia , Caulerpa/química , Microbioma Gastrointestinal/efeitos dos fármacos , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Adjuvantes Imunológicos/sangue , Animais , Biodiversidade , Colo/efeitos dos fármacos , Ciclofosfamida/farmacologia , Ácidos Graxos Voláteis , Imunidade nas Mucosas , Imunoglobulina A Secretora , Interleucina-1beta/sangue , Lactobacillus , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Mucina-2 , RNA Mensageiro/metabolismo , Baço , Timo , Fator de Necrose Tumoral alfa/sangue
12.
Redox Biol ; 26: 101269, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31330482

RESUMO

The gastrointestinal (GI) tract can play a critical role in the development of pathologies associated with overeating, overweight and obesity. We previously observed that supplementation with anthocyanins (AC) (particularly glycosides of cyanidin and delphinidin) mitigated high fat diet (HFD)-induced development of obesity, dyslipidemia, insulin resistance and steatosis in C57BL/6J mice. This paper investigated whether these beneficial effects could be related to AC capacity to sustain intestinal monolayer integrity, prevent endotoxemia, and HFD-associated dysbiosis. The involvement of redox-related mechanisms were further investigated in Caco-2 cell monolayers. Consumption of a HFD for 14 weeks caused intestinal permeabilization and endotoxemia, which were associated with a decreased ileum expression of tight junction (TJ) proteins (occludin, ZO-1 and claudin-1), increased expression of NADPH oxidase (NOX1 and NOX4) and NOS2 and oxidative stress, and activation of redox sensitive signals (NF-κB and ERK1/2) that regulate TJ dynamics. AC supplementation mitigated all these events and increased GLP-2 levels, the intestinal hormone that upregulates TJ protein expression. AC also prevented, in vitro, tumor necrosis factor alpha-induced Caco-2 monolayer permeabilization, NOX1/4 upregulation, oxidative stress, and NF-κB and ERK activation. HFD-induced obesity in mice caused dysbiosis and affected the levels and secretion of MUC2, a mucin that participates in intestinal cell barrier protection and immune response. AC supplementation restored microbiota composition and MUC2 levels and distribution in HFD-fed mice. Thus, AC, particularly delphinidin and cyanidin, can preserve GI physiology in HFD-induced obesity in part through redox-regulated mechanisms. This can in part explain AC capacity to mitigate pathologies, i.e. insulin resistance and steatosis, associated with HFD-associated obesity.


Assuntos
Antocianinas/farmacologia , Dieta Hiperlipídica/efeitos adversos , Microbioma Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/metabolismo , Oxirredução , Substâncias Protetoras/farmacologia , Células CACO-2 , Disbiose , Endotoxemia/tratamento farmacológico , Endotoxemia/etiologia , Endotoxemia/metabolismo , Células Caliciformes/metabolismo , Humanos , Mucina-2/genética , Mucina-2/metabolismo , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Permeabilidade/efeitos dos fármacos , Transdução de Sinais
13.
mBio ; 10(3)2019 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-31213556

RESUMO

Much remains unknown about how the intestinal microbiome interfaces with the protective intestinal mucus layer. Bifidobacterium species colonize the intestinal mucus layer and can modulate mucus production by goblet cells. However, select Bifidobacterium strains can also degrade protective glycans on mucin proteins. We hypothesized that the human-derived species Bifidobacterium dentium would increase intestinal mucus synthesis and expulsion, without extensive degradation of mucin glycans. In silico data revealed that B. dentium lacked the enzymes necessary to extensively degrade mucin glycans. This finding was confirmed by demonstrating that B. dentium could not use naive mucin glycans as primary carbon sources in vitro To examine B. dentium mucus modulation in vivo, Swiss Webster germfree mice were monoassociated with live or heat-killed B. dentium Live B. dentium-monoassociated mice exhibited increased colonic expression of goblet cell markers Krüppel-like factor 4 (Klf4), Trefoil factor 3 (Tff3), Relm-ß, Muc2, and several glycosyltransferases compared to both heat-killed B. dentium and germfree counterparts. Likewise, live B. dentium-monoassociated colon had increased acidic mucin-filled goblet cells, as denoted by Periodic Acid-Schiff-Alcian Blue (PAS-AB) staining and MUC2 immunostaining. In vitro, B. dentium-secreted products, including acetate, were able to increase MUC2 levels in T84 cells. We also identified that B. dentium-secreted products, such as γ-aminobutyric acid (GABA), stimulated autophagy-mediated calcium signaling and MUC2 release. This work illustrates that B. dentium is capable of enhancing the intestinal mucus layer and goblet cell function via upregulation of gene expression and autophagy signaling pathways, with a net increase in mucin production.IMPORTANCE Microbe-host interactions in the intestine occur along the mucus-covered epithelium. In the gastrointestinal tract, mucus is composed of glycan-covered proteins, or mucins, which are secreted by goblet cells to form a protective gel-like structure above the epithelium. Low levels of mucin or alterations in mucin glycans are associated with inflammation and colitis in mice and humans. Although current literature links microbes to the modulation of goblet cells and mucins, the molecular pathways involved are not yet fully understood. Using a combination of gnotobiotic mice and mucus-secreting cell lines, we have identified a human-derived microbe, Bifidobacterium dentium, which adheres to intestinal mucus and secretes metabolites that upregulate the major mucin MUC2 and modulate goblet cell function. Unlike other Bifidobacterium species, B. dentium does not extensively degrade mucin glycans and cannot grow on mucin alone. This work points to the potential of using B. dentium and similar mucin-friendly microbes as therapeutic agents for intestinal disorders with disruptions in the mucus barrier.


Assuntos
Autofagia , Bifidobacterium/fisiologia , Sinalização do Cálcio , Interações entre Hospedeiro e Microrganismos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Animais , Bifidobacterium/enzimologia , Feminino , Microbioma Gastrointestinal , Vida Livre de Germes , Células Caliciformes/fisiologia , Masculino , Camundongos , Mucina-2/genética , Mucinas/metabolismo , Mucinas/farmacologia , Polissacarídeos/metabolismo
14.
Arch Med Res ; 50(1): 2-9, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-31101239

RESUMO

BACKGROUND: Previous studies have reported that nonalcoholic steatohepatitis (NASH) is relevant to intestinal mucosal barrier dysfunction. AIM OF THE STUDY: To investigate the effects of intestinal trefoil factor 3 (TFF3) on intestinal barrier function and endotoxin/toll-like receptor 4(TLR4) expression in NASH rats. METHODS: Sixty NASH rats were divided into control, NASH and NASH-TFF3 treated group. Intestinal permeability, serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), endotoxin (ET), diamine oxidase (DAO) and liver index were examined. HE and PAS staining were performed to observe the histopathology of liver and terminal ileum. Expression of TFF3 and occludin were detected by immunohistochemical staining. mRNA and protein expression of TLR4, nuclear factor-κB (NF-κB), Mucin-2(Muc2) were detected by RT-qPCR and Western Blot. Interleukin (IL) -1ß and IL-10 levels in the ileum were measured by ELISA. RESULTS: In NASH group, levels of AST, ALT, ET, DAO, NAS, liver index and intestinal permeability were higher while occludin expressions were lower than control and NASH-TFF3 treated groups (p <0.05). Histopathology examination showed pathological damages of liver and ileum were alleviated in NASH-TFF3 treated group. NASH-TFF3 treated group had decreased expression levels of TLR4 and NF-κB and increased expression levels of Muc2 than NASH group. Besides, NASH group showed increased IL-1ß and IL-10 levels compared with control group. NASH-TFF3 treated group showed decreased IL-1ß level however increased IL-10 level compared with NASH group. CONCLUSION: Recombinant human TFF3 (rhTFF3) can reduce the expression of TLR4, reduce intestinal permeability, alleviate liver damage and thus may play a therapeutic role in the treatment of NASH rats.


Assuntos
Mucosa Intestinal/patologia , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Hepatopatia Gordurosa não Alcoólica/patologia , Fator Trefoil-3/uso terapêutico , Alanina Transaminase/sangue , Amina Oxidase (contendo Cobre)/sangue , Animais , Aspartato Aminotransferases/sangue , Modelos Animais de Doenças , Endotoxinas/sangue , Humanos , Interleucina-1beta/metabolismo , Masculino , Mucina-2/metabolismo , NF-kappa B/metabolismo , Ocludina/metabolismo , RNA Mensageiro/genética , Ratos , Proteínas Recombinantes/uso terapêutico , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
15.
Arkh Patol ; 81(2): 10-17, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31006774

RESUMO

In the course of the serrated pathway of carcinogenesis, there are changes in the expression of mucins with a characteristic immunophenotypic sign, such as a late loss of intestinal differentiation and an increase in gastric differentiation. OBJECTIVE: To comparatively assess the expression of Muc 2, Muc 5AC, and Muc 6 in hyperplastic polyps (HPs), sessile serrated adenomas (SSAs) and traditional serrated adenomas (TSAs) of the colon for determination of their role in differential diagnosis. MATERIAL AND METHODS: Sixty-five serrated masses from 52 patients were examined. Among them, there were 26 SSAs, 26 HPs, and 13 TSAs. A histological examination was done using hematoxylin and eosin staining; periodic acid-Schiff reaction in combination with alcian blue, as well as immunohistochemistry with anti-Muc 2, anti-Muc 5AC, and anti-Muc 6 antibodies were used. Genetic testing of the specimens for KRAS and BRAF mutations was also carried out. RESULTS: All the serrated neoplasms of the colon exhibited a pronounced expression of Muc 2. A marked Muc 6 expression in the dilated crypt bases was found in 76.9% of SSAs, while no reaction was seen in 92.3% of HPs and in 100% of TSAs. SSAs were characterized by an intense Muc 5AC expression in the whole length of the crypts and in the surface epithelium in contrast with HPs and TSAs, where the expression of the marker was focal. Comparison of the response of the markers and the presence of gene mutations identified that the SSAs with BRAF mutation intensely expressed along the length of the crypt for Muc 5AC and Muc 6; and the TSAs with KRAS mutation had a moderate focal Muc 5AC expression in the crypt bases in 100% of cases. CONCLUSION: For differential diagnosis of the types of serrated adenomas of the colon, it is useful for a pathologist to apply the immunohistochemical markers Muc 2, Muc 5AC, and Muc 6 in his/her practice.


Assuntos
Adenoma , Biomarcadores Tumorais , Neoplasias do Colo , Pólipos do Colo , Mucina-5AC , Mucina-2 , Mucina-6 , Adenoma/metabolismo , Biomarcadores Tumorais/metabolismo , Colo , Neoplasias do Colo/diagnóstico , Pólipos do Colo/diagnóstico , Feminino , Humanos , Imuno-Histoquímica , Masculino , Mucina-5AC/metabolismo , Mucina-2/metabolismo , Mucina-6/metabolismo , Mutação , Proteínas Proto-Oncogênicas B-raf
16.
Surg Today ; 49(11): 887-893, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30879148

RESUMO

Intraductal papillary mucinous neoplasm (IPMN) of the pancreas is characterized by cystic dilation of the pancreatic duct, caused by mucin hypersecretion, with slow progression via the adenoma-carcinoma sequence mechanism. Mutation of GNAS at codon 201 is found exclusively in IPMNs, occurring at a rate of 41-75%. Recent advances in molecular biological techniques have demonstrated that GNAS mutation might play a role in the transformation of IPMNs after the appearance of neoplastic cells, rather than in the tumorigenesis of IPMNs. GNAS mutation is observed frequently in the intestinal subtype of IPMNs with MUC2 expression, and less frequently in IPMNs with concomitant pancreatic ductal adenocarcinoma (PDAC). Research has focused on assessing GNAS mutation status in clinical practice using various samples. In this review, we discuss the clinical application of GNAS mutation assessment to differentiate invasive IPMNs from concomitant PDAC, examine the clonality of recurrent IPMNs in the remnant pancreas using resected specimens, and differentiate pancreatic cystic lesions using cystic fluid collected by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA), duodenal fluid, and serum liquid biopsy samples.


Assuntos
Cromograninas/genética , Subunidades alfa Gs de Proteínas de Ligação ao GTP/genética , Mutação , Neoplasias Primárias Múltiplas , Neoplasias Intraductais Pancreáticas/genética , Neoplasias Pancreáticas/genética , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patologia , Transformação Celular Neoplásica/genética , Códon/genética , Diagnóstico Diferencial , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico , Expressão Gênica , Humanos , Mucina-2/genética , Mucina-2/metabolismo , Neoplasias Intraductais Pancreáticas/diagnóstico , Neoplasias Intraductais Pancreáticas/patologia , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patologia , Proteínas Proto-Oncogênicas p21(ras)
17.
Cancer Chemother Pharmacol ; 83(5): 893-904, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30815720

RESUMO

Irinotecan-induced mucositis is a major oncological problem. Goblet cells secrete mucus, protecting the intestinal mucosa, with secretion altered during mucositis. The enteric nervous system is involved in regulating gut motility and secretion. The aim of this study was to determine whether enteric neural cells and goblet cells are altered following irinotecan treatment. Tumour-bearing Dark Agouti rats were administered a single dose of 175 mg/kg of irinotecan intraperitoneally and 0.01 mg/kg atropine subcutaneously. Experimental and untreated control rats were killed at times 6, 24, 48, 72, 96 and 120 h after treatment. Jejunum and colon samples were formalin fixed. Haematoxylin and eosin staining, Alcian Blue-PAS staining, and immunohistochemistry with S-100 antibody (neural cell marker) were carried out. Statistical analyses were carried out using Kruskal-Wallis test with Dunns post test, Mann Whitney U test, and nonlinear regression. Total goblet cells decreased at 72 h compared with controls in the colon (p < 0.05). The percentage of cavitated goblet cells decreased compared to all other time points at 120 h in the colon. The number of S-100-positive cells in the submucosal plexus decreased in the colon (p = 0.0046) and in the myenteric plexus of the jejunum and colon (p = 0.0058 and p = 0.0022, respectively), on comparing treated with control. Enteric ganglia in the myenteric plexus of the jejunum decreased at 24 h and 96 h. Irinotecan-induced mucositis is associated with increases in mucus secretion and enteric neural cell change. These changes may contribute to the pathophysiology of mucositis through the dysregulation of neural signalling.


Assuntos
Adenocarcinoma/tratamento farmacológico , Enterócitos/efeitos dos fármacos , Irinotecano/efeitos adversos , Neoplasias Mamárias Experimentais/tratamento farmacológico , Mucina-2/metabolismo , Mucina-4/metabolismo , Mucosite/induzido quimicamente , Animais , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Enterócitos/metabolismo , Enterócitos/patologia , Feminino , Células Caliciformes/efeitos dos fármacos , Células Caliciformes/metabolismo , Células Caliciformes/patologia , Irinotecano/uso terapêutico , Jejuno/efeitos dos fármacos , Jejuno/metabolismo , Jejuno/patologia , Mucina-2/genética , Mucina-4/genética , Mucosite/metabolismo , Mucosite/patologia , Ratos Endogâmicos
18.
Br J Cancer ; 120(7): 697-702, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30837681

RESUMO

BACKGROUND: Ampullary adenocarcinoma (AA) originates from either intestinal (INT) or pancreaticobiliary (PB) epithelium. Different prognostic factors of recurrence have been identified in previous studies. METHODS: In 91 AA patients of the AGEO retrospective multicentre cohort, we evaluated the centrally reviewed morphological classification, panel markers of Ang et al. including CK7, CK20, MUC1, MUC2 and CDX2, the 50-gene panel mutational analysis, and the clinicopathological AGEO prognostic score. RESULTS: Forty-three (47%) of the 91 tumours were Ang-INT, 29 (32%) were Ang-PB, 18 (20%) were ambiguous (Ang-AMB) and one could not be classified. Among these 90 tumours, 68.7% of INT tumours were Ang-INT and 78.2% of PB tumours were Ang-PB. MUC5AC expression was detected in 32.5% of the 86 evaluable cases. Among 71 tumours, KRAS, TP53, APC and PIK3CA were the most frequently mutated genes. The KRAS mutation was significantly more frequent in the PB subtype. In multivariate analysis, only AGEO prognostic score and tumour subtype were associated with relapse-free survival. Only AGEO prognostic score was associated with overall survival. CONCLUSIONS: Mutational analysis and MUC5AC expression provide no additional value in the prognostic evaluation of AA patients. Ang et al. classification and the AGEO prognostic score were confirmed as a strong prognosticator for disease recurrence.


Assuntos
Adenocarcinoma/genética , Ampola Hepatopancreática , Neoplasias do Ducto Colédoco/genética , Neoplasias Duodenais/genética , Adenocarcinoma/classificação , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Proteína da Polipose Adenomatosa do Colo/genética , Fator de Transcrição CDX2/metabolismo , Classe I de Fosfatidilinositol 3-Quinases/genética , Neoplasias do Ducto Colédoco/classificação , Neoplasias do Ducto Colédoco/metabolismo , Neoplasias do Ducto Colédoco/patologia , Neoplasias Duodenais/classificação , Neoplasias Duodenais/metabolismo , Neoplasias Duodenais/patologia , Feminino , Humanos , Imuno-Histoquímica , Queratina-20/metabolismo , Queratina-7/metabolismo , Masculino , Pessoa de Meia-Idade , Mucina-5AC/metabolismo , Mucina-1/metabolismo , Mucina-2/metabolismo , Prognóstico , Proteínas Proto-Oncogênicas p21(ras)/genética , Estudos Retrospectivos , Proteína Supressora de Tumor p53/genética
19.
Toxicol Lett ; 309: 1-9, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30904571

RESUMO

Aflatoxin M1 (AFM1), ochratoxin A (OTA), and zearalenone (ZEA) are mycotoxins commonly found in milk. Mycotoxin contamination has caused food safety concerns worldwide since most of the toxic effects in humans are serious. The combined toxic effects of these mycotoxins on intestinal epithelial cells have not been reported. Herein, we investigated the combined effects of AFM1, OTA, and ZEA on intestinal integrity and define the underlying mechanisms(s) of their effects in Caco-2/HT29-MTX co-cultures. Our results showed that the mixtures of AFM1 + OTA, AFM1 + ZEA, and AFM1 + ZEA + OTA significantly increased epithelial permeability. Immunofluorescence analysis and transmission electron microscopy revealed that mycotoxins altered TJ proteins morphology and disrupted their structures. Also, the present study showed that mixtures of mycotoxins significantly modulated MUC5AC and MUC5B mRNA levels and protein secretion. This study demonstrated that the effects of mixtures of mycotoxins on intestinal barrier function were more significant than AFM1 alone. More importantly, the damage of intestinal integrity caused by mycotoxins was correlated to the change of the TJ proteins location and the decrease of mucin secretion. Mixtures of AFM1, OTA, and ZEA in food might pose a health risk to consumers, particularly in children, and toxin risks should be considered.


Assuntos
Aflatoxina M1/toxicidade , Mucosa Intestinal/efeitos dos fármacos , Mucinas/metabolismo , Ocratoxinas/toxicidade , Zearalenona/toxicidade , Células CACO-2 , Técnicas de Cocultura , Contaminação de Alimentos , Células HT29 , Humanos , Mucosa Intestinal/metabolismo , Mucina-5AC/análise , Mucina-5AC/genética , Mucina-2/análise , Mucina-2/genética , Mucina-5B/análise , Mucina-5B/genética , Mucinas/genética , Permeabilidade , RNA Mensageiro/análise , Proteínas de Junções Íntimas/análise
20.
Biomed Pharmacother ; 111: 901-908, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30841469

RESUMO

AIMS: Bisphenol A (BPA) can induce intestinal epithelial cell barrier dysfunction; however, its effects on the intestinal mucus barrier remain unclear. We used LS174T cells as a model to investigate the effects of BPA on the functions of intestinal goblet cells. MAIN METHODS: This study used CCK-8, flow cytometry, ELISA and real-time PCR to investigate the effects of BPA on mitochondrial dynamics, oxidative stress and apoptosis in goblet cells. In addition, mucin synthesis and secretion were evaluated using PAS staining and a PAS assay, respectively. KEY FINDINGS: Our results indicate that BPA reduced cell viability in a time- and concentration-dependent manner. BPA induced mitochondrial dysfunction, as indicated by the depolarization of the mitochondrial membrane potential, inhibition of mitochondrial respiratory chain complex enzyme activity and reduction of ATP production. Moreover, BPA caused oxidative stress by significantly increasing the accumulation of ROS, as well as oxidative stress products, and reducing the antioxidant capacity. Furthermore, BPA induced intestinal goblet cell apoptosis, accompanied by increased DNA fragmentation, caspase-3, -8, -9,-10 gene expression and enzyme activity. Additionally, BPA inhibited the synthesis and secretion of mucin 2. SIGNIFICANCE: Our data suggest that BPA affected the secretory function of intestinal goblet cells by inducing mitochondrial dysfunction, oxidative stress, and apoptosis.


Assuntos
Compostos Benzidrílicos/farmacologia , Células Caliciformes/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Doenças Mitocondriais/tratamento farmacológico , Mucina-2/antagonistas & inibidores , Estresse Oxidativo/efeitos dos fármacos , Fenóis/farmacologia , Trifosfato de Adenosina/metabolismo , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Células Caliciformes/metabolismo , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/metabolismo , Doenças Mitocondriais/metabolismo , Espécies Reativas de Oxigênio/metabolismo
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