RESUMO
Celiac disease (CD) is a food enteropathy that occurs in genetically susceptible individuals following the ingestion of gluten. Both gluten cytotoxicity and immunity activation play a role in CD pathogenesis; however, the chronological assessment of the different pathogenic mechanisms remains elusive. The models developed so far have only partially addressed this issue. Herein, Ab°DQ8 transgenic mice were administered wheat gliadin and indomethacin for 10 days to induce enteropathy. Gliadin-induced alteration of the small intestinal architecture was associated with increased expression of tissue transglutaminase in the lamina propria and a marked hypoxic environment. Enteropathic mice showed activation of innate immunity, featuring an increase of pro-inflammatory IFN-γ and IL-15 mRNAs, as well as CD11c+CD103+, CD11b+CD11c+, and CD11b+CD103+ dendritic cell subsets. However, the temporal assessment of examined parameters indicated that the induction of innate immunity during the generation of the mucosal lesion, occurred belatedly, highlighting a major role of gliadin intrinsic cytotoxicity in the pathogenic mechanism of this model. These results have important implications for the use of this model to test the impact of biotechnological interventions to reduce the cytotoxicity of gliadin.
Assuntos
Doença Celíaca/etiologia , Suscetibilidade a Doenças , Gliadina/imunologia , Antígenos HLA-DQ/genética , Antígenos HLA-DQ/imunologia , Imunidade Inata , Animais , Biomarcadores , Doença Celíaca/metabolismo , Doença Celíaca/patologia , Citocinas/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Expressão Gênica , Humanos , Mediadores da Inflamação/metabolismo , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Intestino Delgado , Camundongos , Camundongos TransgênicosRESUMO
We developed a discriminant method based on the stable isotope ratio of carbon and nitrogen (δ13C and δ15N) to evaluate whether monosodium glutamate (MSG) is used in processed food samples. δ13C measurements were performed by elemental analyzer/isotope-ratio mass spectrometry (EA/IRMS) for on glutamic acid isolated from samples at high purity, and δ15N measurements were performed by gas chromatography/combustion/IRMS (GC/C/IRMS) following the purification and derivatization steps. By applying these methods, the δ13C and δ15N values for glutamic acid present in a wide variety of processed foods were obtained. Subsequently, discriminant analysis, which is a statistical analysis method, was performed by using the δ13C and δ15N values from seasoning MSG and glutamic acid from foodstuffs of known origin, and the discriminant function was derived. By substituting the measured δ13C and δ15N values of processed food samples into this discriminant function and classifying samples into two groups, seasoning MSG (the seasoning group) and glutamic acid in foodstuffs (the foodstuff group), we determined whether seasoning MSG had been used in the processed food samples. As a result, the accuracy of distinguishing between the seasoning group and the foodstuff group was very high, i.e., 96.2%, indicating that the proposed method is a highly robust and accurate method for determining whether seasoning MSG has been used in for processed foods.
RESUMO
Recently, a number of consumers have begun to appreciate more natural ingredients and have become less willing to consume monosodium glutamate (MSG) as a seasoning. By measuring stable isotope ratios (δ13C and δ15N) of glutamic acid contained in foodstuffs and MSG used as seasoning, we attempted to distinguish between both using elemental analyzer-isotope-ratio mass spectrometry (EA/IRMS) and gas chromatography/combustion/IRMS (GC/C/IRMS). As a result, seasoning MSG was observed to have a lower δ15N value than glutamic acid in foodstuffs. We statistically analyzed the stable isotope ratio data using canonical discriminant analysis, thereby differentiating seasoning MSG from foodstuff-derived glutamic acid at an accuracy of 96.7%. This method is effective for distinguishing glutamic acid in foodstuffs from seasoning MSG.
RESUMO
Recent growing health awareness is leading to increasingly conscious decisions by consumers regarding the production and traceability of food. Stable isotopic compositions provide useful information for tracing the origin of foodstuffs and processes of food production. Plants exhibit different ratios of stable carbon isotopes (δ13C) because they utilized different photosynthetic (carbon fixation) pathways and grow in various environments. The origins of glutamic acid in foodstuffs can be differentiated on the basis of these photosynthetic characteristics. Here, we have developed a method to isolate glutamic acid in foodstuffs for determining the δ13C value by elemental analyzer-isotope-ratio mass spectrometry (EA/IRMS) without unintended isotopic fractionation. Briefly, following acid-hydrolysis, samples were defatted and passed through activated carbon and a cation-exchange column. Then, glutamic acid was isolated using preparative HPLC. This method is applicable to measuring, with a low standard deviation, the δ13C values of glutamic acid from foodstuffs derived from C3 and C4 plants and marine algae.
Assuntos
Análise de Alimentos , Ácido Glutâmico/isolamento & purificação , Kelp/química , Phaeophyceae/química , Solanum lycopersicum/química , Zea mays/química , Isótopos de Carbono , Cromatografia Líquida de Alta Pressão , Ácido Glutâmico/química , Espectrometria de MassasRESUMO
We fed rats noodle (N) -diet containing 30 wt.% instant noodle with a 26% fat-to-energy ratio for 30 days (N-group). Compared with rats that were fed the same amount of nutrients (C-group), the N-group showed lower liver triacylglycerol levels and higher fecal cholesterol levels. We then analyzed transcriptome of the hypothalamic-pituitary (HP), the liver and the white adipose tissue (WAT). Thyroid stimulating hormone (Tshb), and its partner, glycoprotein hormone genes were up-regulated in the HP of N-group. Sterol regulatory element binding transcription factors were activated in the liver of N-group, while an up-regulation of the angiogenic signal occurred in the WAT of N-group. N-group showed higher urine noradrenaline (NA) level suggesting that these tissue signals are regulated by NA and Tshb. The N-diet contains 0.326 wt.% glutamate, 0.00236 wt.% 6-shogaol and Maillard reaction products. Our results suggest that these ingredients may affect lipid homeostasis via the HP axis.
Assuntos
Gorduras na Dieta/análise , Crescimento e Desenvolvimento/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Hipófise/efeitos dos fármacos , Tecido Adiposo Branco/efeitos dos fármacos , Tecido Adiposo Branco/metabolismo , Aminoácidos/sangue , Animais , Catecolaminas/urina , Hipotálamo/fisiologia , Masculino , Hipófise/fisiologia , Ratos , Ratos Wistar , Transcriptoma/efeitos dos fármacosRESUMO
The present study investigated the antiallergic and anti-inflammatory effects of 10-hydroxy-cis-12-octadecenoic acid (HYA), a novel gut microbial metabolite of linoleic acid, in NC/Nga mice, a model of atopic dermatitis (AD). Feeding HYA decreased the plasma immunoglobulin E level and skin infiltration of mast cells with a concomitant decrease in dermatitis score. HYA feeding decreased TNF-α and increased claudin-1, a tight junction protein, levels in the mouse skin. Cytokine expression levels in the skin and intestinal Peyer's patches cells suggested that HYA improved the Th1/Th2 balance in mice. Immunoglobulin A concentration in the feces of the HYA-fed mice was approximately four times higher than that in the control mice. Finally, denaturing gradient gel electrophoresis of the PCR-amplified 16 S rRNA gene of fecal microbes indicated the modification of microbiota by HYA. Taken together, the alterations in the intestinal microbiota might be, at least in part, associated with the antiallergic effect of HYA.
Assuntos
Dermatite Atópica/dietoterapia , Suplementos Nutricionais , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/fisiologia , Ácido Linoleico/farmacologia , Ácidos Oleicos/farmacologia , Ração Animal , Animais , Comportamento Animal/efeitos dos fármacos , Citocinas/genética , Citocinas/metabolismo , Dieta/veterinária , Fezes/química , Regulação da Expressão Gênica/fisiologia , Imunoglobulina A/química , Inflamação/tratamento farmacológico , Ácido Linoleico/administração & dosagem , Ácido Linoleico/química , Camundongos , Estrutura Molecular , Ácidos Oleicos/administração & dosagem , Ácidos Oleicos/química , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Intracellular zinc is required for a variety of cell functions, but its precise roles in the maintenance of the intestinal tight junction (TJ) barrier remain unclear. The present study investigated the essential roles of intracellular zinc in the preservation of intestinal TJ integrity and the underlying molecular mechanisms. Depletion of intracellular zinc in both intestinal Caco-2 cells and mouse colons through the application of a cell-permeable zinc chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) induced a disruption of the TJ barrier, as indicated by increased FITC-labeled dextran flux and decreased transepithelial electrical resistance. The TPEN-induced TJ disruption is associated with downregulation of two TJ proteins, occludin and claudin-3. Biotinylation of cell surface proteins revealed that the zinc depletion induced the proteolysis of occludin but not claudin-3. Occludin proteolysis was sensitive to the inhibition of calpain activity, and increased calpain activity was observed in the zinc-depleted cells. Although quantitative PCR analysis and promoter reporter assay have demonstrated that the zinc depletion-induced claudin-3 downregulation occurred at transcriptional levels, a site-directed mutation in the egr1 binding site in the claudin-3 promoter sequence induced loss of both the basal promoter activity and the TPEN-induced decreases. Reduced egr1 expression by a specific siRNA also inhibited claudin-3 expression and transepithelial electrical resistance maintenance in cells. This study shows that intracellular zinc has an essential role in the maintenance of the intestinal epithelial TJ barrier through regulation of occludin proteolysis and claudin-3 transcription.
Assuntos
Claudina-3/metabolismo , Colo/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Ocludina/metabolismo , Sulfato de Zinco/farmacologia , Zinco/metabolismo , Animais , Sítios de Ligação , Células CACO-2 , Quelantes/farmacologia , Claudina-3/genética , Colo/metabolismo , Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Impedância Elétrica , Células Epiteliais/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Masculino , Camundongos Endogâmicos ICR , Permeabilidade , Regiões Promotoras Genéticas , Proteólise , Interferência de RNA , Transdução de Sinais , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo , Transcrição Gênica , Transfecção , Zinco/deficiência , Sulfato de Zinco/metabolismoRESUMO
Metabolic syndrome is characterized by low-grade chronic systemic inflammation, which is associated with intestinal hyperpermeability. This study examined the effects of 3 high-fat diets (HFDs) composed of different fat sources (soybean oil and lard) on the intestinal permeability, tight junction (TJ) protein expression, and cecal bile acid (BA) concentrations in mice, and then analyzed their interrelations. C57/BL6 mice were fed the control diet, HFD (soybean oil), HFD (lard), and HFD (mix; containing equal concentrations of soybean oil and lard) for 8 wk. Glucose tolerance, intestinal permeability, TJ protein expression, and cecal BA concentration were evaluated. Feeding with the 3 HDFs similarly increased body weight, liver weight, and fat pad weight, and induced glucose intolerance and intestinal hyperpermeability. The expression of TJ proteins, zonula occludens-2 and junctional adhesion molecule-A, were lower in the colons of the 3 HFD groups than in the control group (P < 0.05), and these changes appeared to be related to intestinal hyperpermeability. Feeding with HFDs increased total secondary BA (SBA) and total BA concentrations along with increases in some individual BAs in the cecum. Significant positive correlations between intestinal permeability and the concentrations of most SBAs, such as deoxycholic acid and ω-muricholic acids, were detected (P < 0.05). These results suggest that the HFD-induced intestinal hyperpermeability is associated with increased BA secretion. The abundance of SBAs in the large intestine may be responsible for the hyperpermeability.
Assuntos
Ácidos e Sais Biliares/metabolismo , Dieta Hiperlipídica/efeitos adversos , Gorduras na Dieta/efeitos adversos , Enteropatias/etiologia , Intestino Grosso/patologia , Síndrome Metabólica/patologia , Junções Íntimas/metabolismo , Tecido Adiposo/metabolismo , Animais , Ceco/metabolismo , Ácidos Cólicos/metabolismo , Colo/metabolismo , Ácido Desoxicólico/metabolismo , Gorduras na Dieta/administração & dosagem , Inflamação/metabolismo , Enteropatias/metabolismo , Intestino Grosso/metabolismo , Moléculas de Adesão Juncional/metabolismo , Fígado , Masculino , Camundongos Endogâmicos C57BL , Óleo de Soja , Aumento de Peso , Proteína da Zônula de Oclusão-2/metabolismoRESUMO
Obesity is one of the major health problems throughout the world. The present study investigated the preventive effect of epilactose--a rare non-digestible disaccharide--on obesity and metabolic disorders in mice fed high-fat (HF) diets. Feeding with HF diets increased body weight gain, fat pad weight and adipocyte size in mice (P<0·01), and these increases were effectively prevented by the use of supplemental epilactose without influencing food intake (P<0·01). Caecal pools of SCFA such as acetic and propionic acids in mice fed epilactose were higher compared with mice not receiving epilactose. Supplemental epilactose increased the expression of uncoupling protein (UCP)-1, which enhances energy expenditure, to 2-fold in the gastrocnemius muscle (P=0·04) and to 1·3-fold in the brown adipose tissue (P=0·02) in mice fed HF diets. Feeding HF diets induced pro-inflammatory macrophage infiltration into white adipose tissue, as indicated by the increased expression of monocyte chemotactic protein-1, TNF-α and F4/80, and these increases were attenuated by supplemental epilactose. In differentiated myogenic-like C2C12 cells, propionic acid, but not acetic or n-butyric acids, directly enhanced UCP-1 expression by approximately 2-fold (P<0·01). Taken together, these findings indicate that the epilactose-mediated increase in UCP-1 in the skeletal muscle and brown adipose tissue can enhance whole-body energy expenditure, leading to effective prevention of obesity and metabolic disorders in mice fed HF diets. It is suggested that propionic acid--a bacterial metabolite--acts as a mediator to induce UCP-1 expression in skeletal muscles.
Assuntos
Fármacos Antiobesidade/uso terapêutico , Dissacarídeos/uso terapêutico , Canais Iônicos/agonistas , Proteínas Mitocondriais/agonistas , Músculo Esquelético/metabolismo , Obesidade/prevenção & controle , Prebióticos , Regulação para Cima , Tecido Adiposo Marrom/imunologia , Tecido Adiposo Marrom/metabolismo , Tecido Adiposo Marrom/patologia , Tecido Adiposo Branco/imunologia , Tecido Adiposo Branco/metabolismo , Tecido Adiposo Branco/patologia , Animais , Anti-Inflamatórios não Esteroides/metabolismo , Anti-Inflamatórios não Esteroides/uso terapêutico , Fármacos Antiobesidade/metabolismo , Linhagem Celular , Dieta Hiperlipídica/efeitos adversos , Dissacarídeos/metabolismo , Metabolismo Energético , Fermentação , Microbioma Gastrointestinal , Canais Iônicos/genética , Canais Iônicos/metabolismo , Ativação de Macrófagos , Masculino , Camundongos Endogâmicos C57BL , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Músculo Esquelético/imunologia , Obesidade/imunologia , Obesidade/metabolismo , Obesidade/microbiologia , Propionatos/metabolismo , Distribuição Aleatória , Proteína Desacopladora 1RESUMO
Secretory immunoglobulin A (IgA) mediates the mucosal immune system, which provides the first line of defense against inhaled and ingested pathogenic bacteria and viruses. Lactobacillus plantarum AYA increases the IgA level of Peyer's patch (PP) cells, but the recommended amount of consumption and the mechanism of action remains unclear. Better understanding of these is essential to development of L. plantarum AYA for use in functional foods. Therefore, we investigated the dose-response effect (in vivo) and mechanism (in vitro) of IgA enhancement induced by L. plantarum AYA. In the small intestine of the mice fed a diet containing 0.03% or 0.3% of L. plantarum AYA powder for 4 weeks, the IgA levels were significantly increased. Thus, it is suggested that the recommended amount of consumption of L. plantarum AYA is about 0.72â mg per day. In addition, the bacterial cell wall fraction significantly enhanced the IgA production level of murine PP cells in the in vitro assay. The ability of whole cells and the cell wall fraction to enhance IgA levels was significantly inhibited by an anti-Toll-like receptor-2 (TLR-2) antibody, which suggests that the cell wall fraction of L. plantarum AYA increases the IgA level via TLR-2. These findings indicate that L. plantarum AYA is a potential functional food source that maintains mucosal immunity.
RESUMO
We isolated Bifidobacterium kashiwanohense JCM 15439 from the feces of a healthy Japanese infant and proposed it as the type strain of a novel species within the genus Bifidobacterium. Here, we report the complete genome sequence of this organism.
RESUMO
Gut microbiota and probiotic strains play an important role in oral tolerance by modulating regulatory and effector cell components of the immune system. We have previously described the ability of Lactobacilli to influence both the innate and adaptive immunity to wheat gluten, a food antigen, in mouse. In this study, we further explored the immunomodulatory mechanisms elicited in this model by testing three specific probiotic strains, namely L. rhamnosus OLL2838, B. infantis ATCC15697 and S. thermophilus Sfi39. In vitro analysis showed the all tested strains induced maturation of bone marrow derived dendritic cells (DCs). However, only L. rhamnosus induced appreciable levels of IL-10 and nitric oxide productions, whereas S. thermophilus essentially elicited IL-12 and TNF-α. The anti-inflammatory ability of OLL2838 was then tested in vivo by adopting mice that develop a gluten-specific enteropathy. This model is characterized by villus blunting, crypt hyperplasia, high levels of intestinal IFN-γ, increased cell apoptosis in lamina propria, and reduced intestinal total glutathione (GSHtot) and glutathione S-transferase (GST) activity. We found that, following administration of OLL2838, GSHtot and GST activity were enhanced, whereas caspase-3 activity was reduced. On the contrary, this probiotic strain failed in recovering the normal histology and further increased intestinal IFN-γ. Confocal microscopy revealed the inability of the probiotic strain to appropriately interact with enterocytes of the small intestine and with Peyer's patches in treated mice. In conclusion, these data highlighted the potential of L. rhamnosus OLL2838 to recover specific toxicity parameters induced by gluten in enteropathic mice through mechanisms that involve induction of low levels of reactive oxygen species (ROS).
Assuntos
Imunomodulação , Enteropatias/imunologia , Enteropatias/microbiologia , Lacticaseibacillus rhamnosus/imunologia , Probióticos , Animais , Antígenos CD/metabolismo , Apoptose , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Antígenos HLA-DQ/genética , Antígenos HLA-DQ/imunologia , Humanos , Imunidade Inata , Imunofenotipagem , Enteropatias/metabolismo , Intestino Delgado/imunologia , Intestino Delgado/metabolismo , Intestino Delgado/microbiologia , Camundongos , Camundongos Transgênicos , Óxido Nítrico , Estresse OxidativoRESUMO
Gut microbial metabolites of polyunsaturated fatty acids have attracted much attention because of their various physiological properties. Dysfunction of tight junction (TJ) in the intestine contributes to the pathogenesis of many disorders such as inflammatory bowel disease. We evaluated the effects of five novel gut microbial metabolites on tumor necrosis factor (TNF)-α-induced barrier impairment in Caco-2 cells and dextran sulfate sodium-induced colitis in mice. 10-Hydroxy-cis-12-octadecenoic acid (HYA), a gut microbial metabolite of linoleic acid, suppressed TNF-α and dextran sulfate sodium-induced changes in the expression of TJ-related molecules, occludin, zonula occludens-1, and myosin light chain kinase. HYA also suppressed the expression of TNF receptor 2 (TNFR2) mRNA and protein expression in Caco-2 cells and colonic tissue. In addition, HYA suppressed the protein expression of TNFR2 in murine intestinal epithelial cells. Furthermore, HYA significantly up-regulated G protein-coupled receptor (GPR) 40 expression in Caco-2 cells. It also induced [Ca(2+)]i responses in HEK293 cells expressing human GPR40 with higher sensitivity than linoleic acid, its metabolic precursor. The barrier-recovering effects of HYA were abrogated by a GPR40 antagonist and MEK inhibitor in Caco-2 cells. Conversely, 10-hydroxyoctadacanoic acid, which is a gut microbial metabolite of oleic acid and lacks a carbon-carbon double bond at Δ12 position, did not show these TJ-restoring activities and down-regulated GPR40 expression. Therefore, HYA modulates TNFR2 expression, at least partially, via the GPR40-MEK-ERK pathway and may be useful in the treatment of TJ-related disorders such as inflammatory bowel disease.
Assuntos
Células Epiteliais/metabolismo , Intestinos/citologia , Ácido Linoleico/metabolismo , Ácidos Oleicos/metabolismo , Animais , Células CACO-2 , Colite/metabolismo , Células Epiteliais/citologia , Feminino , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Sistema de Sinalização das MAP Quinases/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Receptores Acoplados a Proteínas G/metabolismoRESUMO
We previously isolated the commensal bacteria lactobacilli and bifidobacteria from the Thoroughbred intestine and prepared the horse probiotics LacFi(TM), consisting of Lactobacillus ruminis KK14, L. equi KK 15, L. reuteri KK18, L. johnsonii KK21, and Bifidobacterium boum HU. Here, we found that the five LacFi(TM) constituent strains remarkably suppressed pro-inflammatory interleukin-17 production in mouse splenocytes stimulated with interleukin-6 and transforming growth factor-ß. The protective effects of the probiotic on impaired intestinal barrier function were evaluated in Caco-2 cells treated with tumor necrosis factor-α. Evaluation of transepithelial resistance showed that all the strains exhibited intestinal barrier protective activity, with significant suppression of barrier impairment by L. reuteri KK18. The LacFi(TM) constituent strains were detected in neonatal LacFi(TM)-administered Thoroughbred feces using polymerase chain reaction denaturing gradient gel electrophoresis and culture methods. These five strains were found to be the predominant lactobacilli and bifidobacteria in the intestinal microbiota of LacFi(TM)-administered Thoroughbreds. Administration of LacFi(TM) to neonatal Thoroughbreds decreased diarrhea incidence from 75.9% in the control group (n=29 neonatal Thoroughbreds) to 30.7% in the LacFi(TM)-administered group (n=101 neonatal Thoroughbreds) immediately after birth to 20 weeks after birth. LacFi(TM) treatment also prevented diarrhea especially at and around 4 weeks and from 10 to 16 weeks. The duration of diarrhea was also shorter in the probiotics-administered group (7.4 ± 0.8 days) than in the control group (14.0 ± 3.2 days). These results indicate that the LacFi(TM) probiotics regulates intestinal function and contributes to diarrhea prevention.
RESUMO
Bifidobacterial plasmids reported so far are derived from a limited number of strains and plasmids of bifidobacterial type strains isolated from humans are unknown. We found that Bifidobacterium kashiwanohenseâ JCM 15439 (type strain) isolated from a healthy infant contained two cryptic plasmids, designated pBBKW-1 and pBBKW-2. We determined and analyzed the complete sequences of both plasmids. pBBKW-1 (7716 bp) was predicted to replicate by a rolling-circle mechanism and encode six protein-coding genes, two of which are putative replication proteins. pBBKW-1 seems to be a cointegrate plasmid containing two copies of the plasmid pMG1 from Bifidobacterium longum. pBBKW-2 (2920 bp) was predicted to encode six protein-coding genes and be a theta-type replicating plasmid, which has been reported to be more stable than a rolling circle-type replicating plasmid frequently found in bifidobacteria. Our finding will provide new insights into safe recombinant plasmid constructions for humans.
Assuntos
Sequência de Bases , Bifidobacterium/genética , Fezes/microbiologia , Plasmídeos/genética , Análise de Sequência de DNA , Bifidobacterium/isolamento & purificação , Humanos , Lactente , Dados de Sequência MolecularRESUMO
BACKGROUND: Probiotics species appear to differentially regulate the intestinal immune response. Moreover, we have shown that different immune-modulatory abilities can be found among probiotic strains belonging to the same species. In this study, we further addressed this issue while studying L. gasseri, a species that induces relevant immune activities in human patients. RESULTS: We determined the ability of two strains of L. gasseri, OLL2809 and L13-Ia, to alter cell surface antigen expression, cytokine production and nuclear erythroid 2-related factor 2 (Nrf2)-mediated cytoprotection in murine bone marrow-derived dendritic cells (DCs) and MODE-K cells, which represent an enterocyte model. Differential effects of L. gasseri strains were observed on the expression of surface markers in mature DCs; nevertheless, both strains dramatically induced production of IL-12, TNF-α and IL-10. Distinctive responses to OLL2809 and L13-Ia were also shown in MODE-K cells by analyzing the expression of MHC II molecules and the secretion of IL-6; however, both L. gasseri strains raised intracellular glutathione. Treatment of immature DCs with culture medium from MODE-K monolayers improved cytoprotection and modified the process of DC maturation by down-regulating the expression of co-stimulatory markers and by altering the cytokine profile. Notably, bacteria-conditioned MODE-K cell medium suppressed the expression of the examined cytokines, whereas cytoprotective defenses were significantly enhanced only in DCs exposed to OLL2809-conditioned medium. These effects were essentially mediated by secreted bacterial metabolites. CONCLUSIONS: We have demonstrated that L. gasseri strains possess distinctive abilities to modulate in vitro DCs and enterocytes. In particular, our results highlight the potential of metabolites secreted by L. gasseri to influence enterocyte-DC crosstalk. Regulation of cellular mechanisms of innate immunity by selected probiotic strains may contribute to the beneficial effects of these bacteria in gut homeostasis.
Assuntos
Imunidade Inata , Fatores Imunológicos/farmacologia , Lactobacillus/imunologia , Probióticos/farmacologia , Animais , Antígenos de Superfície/metabolismo , Células Cultivadas , Citocinas/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/microbiologia , Enterócitos/imunologia , Enterócitos/microbiologia , Camundongos , Fator 2 Relacionado a NF-E2/metabolismoRESUMO
Although some bacterial strains show potential to prevent colitis, their mechanisms are not fully understood. Here, we investigated the anti-colitic mechanisms of Bifidobacterium longum subsp. infantis JCM 1222(T), focusing on the relationship between interleukin (IL)-17A secreting CD4(+) T cells and intestinal epithelial costimulatory molecules in mice. Oral administration of JCM 1222(T) to mice alleviated dextran sulfate sodium (DSS)-induced acute colitis. The expression of type 1 helper T (Th1)- and IL-17 producing helper T (Th17)-specific cytokines and transcriptional factors was suppressed by JCM 1222(T) treatment. Intestinal epithelial cells (IECs) from colitic mice induced IL-17A production from CD4(+) T cells in a cell-cell contact-dependent manner, and this was suppressed by oral treatment with JCM 1222(T). Using blocking antibodies for costimulatory molecules, we revealed that epithelial costimulatory molecules including CD80 and CD40, which were highly expressed in IECs from colitic mice, were involved in IEC-induced IL-17A response. Treatment of mice and intestinal epithelial cell line Colon-26 cells with JCM 1222(T) decreased the expression of CD80 and CD40. Collectively, these data indicate that JCM 1222(T) negatively regulate epithelial costimulatory molecules, and this effect might be attributed, at least in part, to suppression of IL-17A in DSS-induced colitis.
Assuntos
Bifidobacterium/metabolismo , Colite/metabolismo , Interleucina-17/metabolismo , Animais , Antígeno B7-1/genética , Antígeno B7-1/metabolismo , Antígeno B7-2/genética , Antígeno B7-2/metabolismo , Antígenos CD40/genética , Antígenos CD40/metabolismo , Colite/induzido quimicamente , Colite/genética , Colite/imunologia , Colite/microbiologia , Citocinas/metabolismo , Sulfato de Dextrana/efeitos adversos , Feminino , Regulação da Expressão Gênica , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Camundongos , Células Th17/imunologia , Células Th17/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
Probiotics and gut microbiota have a significant impact on gut homeostasis in the host. Recent clinical studies demonstrated the ameliorative features of several kinds of probiotics in intestinal disorders, such as inflammatory bowel diseases (IBDs). Interleukin (IL)-17 is a potent inflammatory cytokine, and T-helper (Th)17 cells and other IL-17-producing cells are involved in the pathogenesis of IBD. Multiple mechanisms of action have been suggested to explain the protective anti-inflammatory effects of probiotics in intestinal inflammation, including the immunoregulation and suppression of Th17 activity and IL-17 production in part by signaling through pattern-recognition receptors such as Toll-like receptor family. However, steady-state Th17 cells have an important role in host defense against fungi and bacteria. Interestingly, recent studies revealed that specific commensal bacterial species such as segmented filamentous bacteria (SFB) induce the accumulation of Th17 cells in the small intestine in many species, including mice. It is important to determine the mechanisms by which intestinal Th17 cells are induced by SFB and whether these or other bacteria with similar properties are present in the human intestine. This brief review focuses on the interaction between probiotics/microbiota and Th17 cells during inflammation (war) and during steady-state homeostatic regulation (peace).
Assuntos
Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Microbiota , Probióticos , Células Th17/imunologia , Animais , Citocinas/metabolismo , Trato Gastrointestinal/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunoglobulina A Secretora/imunologia , Mediadores da Inflamação/metabolismo , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/microbiologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Células Th17/metabolismoRESUMO
SCOPE: We have previously reported that naringenin promotes the tight junction (TJ) integrity in intestinal Caco-2 cells. This study investigated the naringenin-mediated effect in Caco-2 cells with a particular focus on the modulation of TJ structure and claudin-4 transcriptional regulation. METHODS AND RESULTS: Naringenin (10~100 µM) dose-dependently enhanced TJ barrier integrity of Caco-2 cells, indicated by transepithelial electrical resistance and FITC-dextran flux. Immunoblot analysis showed that naringenin increased the cytoskeletal association of TJ proteins, zonula occludens-2, occludin, claudin-1, and claudin-4, simultaneously with increased occludin phosphorylation. The total expression of claudin-4 was also increased by naringenin. Quantitative RT-PCR and luciferase reporter assay revealed that naringenin transcriptionally increased the claudin-4 expression with activation of claudin-4 promoter. The mutation of the binding site of the transcriptional factor Sp1 in the claudin-4 promoter sequence and the pharmacological inhibition of Sp1 partially suppressed the naringenin-mediated activation of the claudin-4 promoter. Further, naringenin induced the heat shock protein 70 expression in the cells. CONCLUSION: Naringenin enhances barrier integrity through the assembly and expression of TJ proteins in intestinal epithelial cells. Naringenin-mediated claudin-4 expression occurs, at least partially, through Sp1-dependent transcriptional regulation. The induction of heat shock protein 70 may be also involved in the increased claudin-4 expression.
Assuntos
Citoesqueleto/metabolismo , Flavanonas/farmacologia , Regulação da Expressão Gênica , Intestinos/efeitos dos fármacos , Proteínas de Junções Íntimas/metabolismo , Células CACO-2 , Claudina-1/genética , Claudina-1/metabolismo , Claudina-4/genética , Claudina-4/metabolismo , Dextranos , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Mucosa Intestinal/citologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Ocludina/genética , Ocludina/metabolismo , Regiões Promotoras Genéticas , Fator de Transcrição Sp1/genética , Fator de Transcrição Sp1/metabolismo , Proteínas de Junções Íntimas/genética , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo , Proteína da Zônula de Oclusão-2/genética , Proteína da Zônula de Oclusão-2/metabolismoRESUMO
We investigated the effects of a chicken collagen hydrolysate (CCH) on the circulation system in humans. A total of 58 subjects with either mild hypertension (systolic blood pressure (SBP) of 140-159 mmHg or diastolic blood pressure (DBP) 90-99 mmHg) or high-normal blood pressure (SBP 130-139 mmHg or DBP 85-89 mmHg) were assigned to two groups, one involving a placebo and the other, the test food (including CCH of 2.9 g/d). The parameters related to each subject's circulation system were monitored over the study period of 18 weeks. The Δbrachial-ankle pulse wave velocity (baPWV), an indicator of arterial stiffness and marker of vascular damage, was significantly lower in the test food group than in the placebo group during the treatment period. The blood pressure in the test food group was also significantly lower than that in the placebo group, while the serum nitrogen oxide was higher in the test food group after the treatment. These results suggest that CCH exerted modulatory effects on the human circulation system.