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1.
Nat Commun ; 12(1): 5958, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34645820

RESUMO

Understanding the functional potential of the gut microbiome is of primary importance for the design of innovative strategies for allergy treatment and prevention. Here we report the gut microbiome features of 90 children affected by food (FA) or respiratory (RA) allergies and 30 age-matched, healthy controls (CT). We identify specific microbial signatures in the gut microbiome of allergic children, such as higher abundance of Ruminococcus gnavus and Faecalibacterium prausnitzii, and a depletion of Bifidobacterium longum, Bacteroides dorei, B. vulgatus and fiber-degrading taxa. The metagenome of allergic children shows a pro-inflammatory potential, with an enrichment of genes involved in the production of bacterial lipo-polysaccharides and urease. We demonstrate that specific gut microbiome signatures at baseline can be predictable of immune tolerance acquisition. Finally, a strain-level selection occurring in the gut microbiome of allergic subjects is identified. R. gnavus strains enriched in FA and RA showed lower ability to degrade fiber, and genes involved in the production of a pro-inflammatory polysaccharide. We demonstrate that a gut microbiome dysbiosis occurs in allergic children, with R. gnavus emerging as a main player in pediatric allergy. These findings may open new strategies in the development of innovative preventive and therapeutic approaches. Trial: NCT04750980.


Assuntos
Alérgenos/imunologia , Hipersensibilidade Alimentar/microbiologia , Microbioma Gastrointestinal/imunologia , Tolerância Imunológica , Hipersensibilidade Respiratória/microbiologia , Alérgenos/efeitos adversos , Animais , Bacteroides/isolamento & purificação , Bacteroides/metabolismo , Bifidobacterium longum/isolamento & purificação , Bifidobacterium longum/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , Clostridiales/isolamento & purificação , Clostridiales/metabolismo , Alérgenos Animais/efeitos adversos , Alérgenos Animais/imunologia , Ovos/efeitos adversos , Faecalibacterium prausnitzii/isolamento & purificação , Faecalibacterium prausnitzii/metabolismo , Feminino , Hipersensibilidade Alimentar/etiologia , Hipersensibilidade Alimentar/imunologia , Humanos , Lipopolissacarídeos/biossíntese , Masculino , Leite/efeitos adversos , Leite/imunologia , Nozes/efeitos adversos , Nozes/imunologia , Pólen/química , Pólen/imunologia , Prunus persica/química , Prunus persica/imunologia , Pyroglyphidae/química , Pyroglyphidae/imunologia , Hipersensibilidade Respiratória/etiologia , Hipersensibilidade Respiratória/imunologia , Urease/biossíntese
2.
mBio ; 12(3): e0362820, 2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34061597

RESUMO

ß-Mannans are hemicelluloses that are abundant in modern diets as components in seed endosperms and common additives in processed food. Currently, the collective understanding of ß-mannan saccharification in the human colon is limited to a few keystone species, which presumably liberate low-molecular-weight mannooligosaccharide fragments that become directly available to the surrounding microbial community. Here, we show that a dominant butyrate producer in the human gut, Faecalibacterium prausnitzii, is able to acquire and degrade various ß-mannooligosaccharides (ß-MOS), which are derived by the primary mannanolytic activity of neighboring gut microbiota. Detailed biochemical analyses of selected protein components from their two ß-MOS utilization loci (F. prausnitzii ß-MOS utilization loci [FpMULs]) supported a concerted model whereby the imported ß-MOS are stepwise disassembled intracellularly by highly adapted enzymes. Coculturing experiments of F. prausnitzii with the primary degraders Bacteroides ovatus and Roseburia intestinalis on polymeric ß-mannan resulted in syntrophic growth, thus confirming the high efficiency of the FpMULs' uptake system. Genomic comparison with human F. prausnitzii strains and analyses of 2,441 public human metagenomes revealed that FpMULs are highly conserved and distributed worldwide. Together, our results provide a significant advance in the knowledge of ß-mannan metabolism and the degree to which its degradation is mediated by cross-feeding interactions between prominent beneficial microbes in the human gut. IMPORTANCE Commensal butyrate-producing bacteria belonging to the Firmicutes phylum are abundant in the human gut and are crucial for maintaining health. Currently, insight is lacking into how they target otherwise indigestible dietary fibers and into the trophic interactions they establish with other glycan degraders in the competitive gut environment. By combining cultivation, genomic, and detailed biochemical analyses, this work reveals the mechanism enabling F. prausnitzii, as a model Ruminococcaceae within Firmicutes, to cross-feed and access ß-mannan-derived oligosaccharides released in the gut ecosystem by the action of primary degraders. A comprehensive survey of human gut metagenomes shows that FpMULs are ubiquitous in human populations globally, highlighting the importance of microbial metabolism of ß-mannans/ß-MOS as a common dietary component. Our findings provide a mechanistic understanding of the ß-MOS utilization capability by F. prausnitzii that may be exploited to select dietary formulations specifically boosting this beneficial symbiont, and thus butyrate production, in the gut.


Assuntos
Faecalibacterium prausnitzii/genética , Faecalibacterium prausnitzii/metabolismo , Microbioma Gastrointestinal/genética , Mananas/metabolismo , Oligossacarídeos/metabolismo , Bacteroides/genética , Bacteroides/metabolismo , Clostridiales/genética , Clostridiales/metabolismo , Colo/microbiologia , Dieta , Faecalibacterium prausnitzii/crescimento & desenvolvimento , Microbioma Gastrointestinal/fisiologia , Humanos , Mananas/classificação , Metagenômica
4.
Methods Mol Biol ; 2167: 13-24, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32712912

RESUMO

Self-cleaving ribozymes are RNA molecules that catalyze a site-specific self-scission reaction. Analysis of self-cleavage is a crucial aspect of the biochemical study and understanding of these molecules. Here we describe a co-transcriptional assay that allows the analysis of self-cleaving ribozymes in different reaction conditions and in the presence of desired ligands and/or cofactors. Utilizing a standard T7 RNA polymerase in vitro transcription system under limiting Mg2+ concentration, followed by a 25-fold dilution of the reaction in desired conditions of self-cleavage (buffer, ions, ligands, pH, temperature, etc.) to halt the synthesis of new RNA molecules, allows the study of self-scission of these molecules without the need for purification or additional preparation steps, such as refolding procedures. Furthermore, because the transcripts are not denatured, this assay likely yields RNAs in conformations relevant to co-transcriptionally folded species in vivo.


Assuntos
Proteínas de Bactérias/metabolismo , RNA Polimerases Dirigidas por DNA/metabolismo , Ensaios Enzimáticos/métodos , Faecalibacterium prausnitzii/metabolismo , Magnésio/metabolismo , RNA Catalítico/metabolismo , Transcrição Genética , Proteínas Virais/metabolismo , Proteínas de Bactérias/genética , Catálise , Eletroforese em Gel de Poliacrilamida , Faecalibacterium prausnitzii/enzimologia , Faecalibacterium prausnitzii/genética , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Íons/química , Cinética , Ligantes , Magnésio/química , Fosfoglucomutase/metabolismo , RNA Catalítico/genética
5.
Circ Res ; 127(4): 453-465, 2020 07 31.
Artigo em Inglês | MEDLINE | ID: mdl-32354259

RESUMO

RATIONALE: The elderly experience profound systemic responses after stroke, which contribute to higher mortality and more severe long-term disability. Recent studies have revealed that stroke outcomes can be influenced by the composition of gut microbiome. However, the potential benefits of manipulating the gut microbiome after injury is unknown. OBJECTIVE: To determine if restoring youthful gut microbiota after stroke aids in recovery in aged subjects, we altered the gut microbiome through young fecal transplant gavage in aged mice after experimental stroke. Further, the effect of direct enrichment of selective bacteria producing short-chain fatty acids (SCFAs) was tested as a more targeted and refined microbiome therapy. METHODS AND RESULTS: Aged male mice (18-20 months) were subjected to ischemic stroke by middle cerebral artery occlusion. We performed fecal transplant gavage 3 days after middle cerebral artery occlusion using young donor biome (2-3 months) or aged biome (18-20 months). At day 14 after stroke, aged stroke mice receiving young fecal transplant gavage had less behavioral impairment, and reduced brain and gut inflammation. Based on data from microbial sequencing and metabolomics analysis demonstrating that young fecal transplants contained much higher SCFA levels and related bacterial strains, we selected 4 SCFA-producers (Bifidobacterium longum, Clostridium symbiosum, Faecalibacterium prausnitzii, and Lactobacillus fermentum) for transplantation. These SCFA-producers alleviated poststroke neurological deficits and inflammation, and elevated gut, brain and plasma SCFA concentrations in aged stroke mice. CONCLUSIONS: This is the first study suggesting that the poor stroke recovery in aged mice can be reversed via poststroke bacteriotherapy following the replenishment of youthful gut microbiome via modulation of immunologic, microbial, and metabolomic profiles in the host.


Assuntos
Ácidos Graxos Voláteis/biossíntese , Transplante de Microbiota Fecal , Microbioma Gastrointestinal/fisiologia , Infarto da Artéria Cerebral Média/terapia , AVC Isquêmico/terapia , Fatores Etários , Animais , Bifidobacterium longum/metabolismo , Química Encefálica , Clostridium symbiosum/metabolismo , Faecalibacterium prausnitzii/metabolismo , Ácidos Graxos Voláteis/análise , Ácidos Graxos Voláteis/sangue , Fezes/química , Interleucina-17/biossíntese , Intestinos/química , Linfócitos Intraepiteliais/fisiologia , Lactobacillus fermentum/metabolismo , Masculino , Camundongos , Mucina-2/metabolismo , Mucina-4/metabolismo , Linfócitos T Reguladores/fisiologia
6.
PLoS One ; 15(1): e0227373, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31910227

RESUMO

METHODS: Patients transplanted at our institution provided fecal samples before, and 3-9 months after KT. Fecal bacterial DNA was extracted and 9 bacteria or bacterial groups were quantified by qPCR. RESULTS: 50 patients (19 controls without diabetes, 15 who developed New Onset Diabetes After Transplantation, NODAT, and 16 with type 2 diabetes before KT) were included. Before KT, Lactobacillus sp. tended to be less frequently detected in controls than in those who would become diabetic following KT (NODAT) and in initially diabetic patients (60%, 87.5%, and 100%, respectively, p = 0.08). The relative abundance of Faecalibacterium prausnitzii was 30 times lower in initially diabetic patients than in controls (p = 0.002). The relative abundance of F. prausnitzii of NODAT patients was statistically indistinguishable from controls and from diabetic patients. The relative abundance of Lactobacillus sp. increased following KT in NODAT and in initially diabetic patients (20-fold, p = 0.06, and 25-fold, p = 0.02, respectively). In contrast, the proportion of Akkermansia muciniphila decreased following KT in NODAT and in initially diabetic patients (2,500-fold, p = 0.04, and 50,000-fold, p<0.0001, respectively). The proportion of Lactobacillus and A. muciniphila did not change in controls between before and after the transplantation. Consequently, after KT the relative abundance of Lactobacillus sp. was 25 times higher (p = 0.07) and the relative abundance of A. muciniphila was 2,000 times lower (p = 0.002) in diabetics than in controls. CONCLUSION: An alteration of the gut microbiota composition involving Lactobacillus sp., A. muciniphila and F. prausnitzii is associated with the glycemic status in KT recipients, raising the question of their role in the genesis of NODAT.


Assuntos
DNA Bacteriano/genética , Diabetes Mellitus Tipo 2/microbiologia , Microbioma Gastrointestinal/genética , Transplante de Rim/efeitos adversos , Akkermansia , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patologia , Faecalibacterium prausnitzii/genética , Faecalibacterium prausnitzii/isolamento & purificação , Faecalibacterium prausnitzii/metabolismo , Fezes/microbiologia , Feminino , Humanos , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Verrucomicrobia/genética , Verrucomicrobia/isolamento & purificação , Verrucomicrobia/metabolismo
7.
J Diabetes ; 12(3): 224-236, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31503404

RESUMO

BACKGROUND: Impaired intestinal barrier structure and function have been validated as an important pathogenic process in type 2 diabetes mellitus (T2DM). Gut dysbiosis is thought to be the critical factor in diabetic intestinal pathogenesis. As the most abundant commensal bacteria, Faecalibacterium prausnitzii (F. prausnitzii) play important roles in gut homeostasis. The microbial anti-inflammatory molecule (MAM), an F. prausnitzii metabolite, has anti-inflammatory potential in inflammatory bowel disease (IBD). Thus, we aimed to explore the function and mechanism of MAM on the diabetic intestinal epithelium. METHODS: 16S high-throughput sequencing was used to analyze the gut microbiota of db/db mice (T2DM mouse model). We transfected a FLAG-tagged MAM plasmid into human colonic cells to explore the protein-protein interactions and observe cell monolayer permeability. For in vivo experiments, db/db mice were supplemented with recombinant His-tagged MAM protein from E. coli BL21 (DE3). RESULTS: The abundance of F. prausnitzii was downregulated in the gut microbiota of db/db mice. Immunoprecipitation (IP) and mass spectroscopy (MS) analyses revealed that MAM potentially interacts with proteins in the tight junction pathway, including zona occludens 1 (ZO-1). FLAG-tagged MAM plasmid transfection stabilized the cell permeability and increased ZO-1 expression in NCM460, Caco2, and HT-29 cells. The db/db mice supplemented with recombinant His-tagged MAM protein showed restored intestinal barrier function and elevated ZO-1 expression. CONCLUSIONS: Our study shows that MAM from F. prausnitzii can restore the intestinal barrier structure and function in DM conditions via the regulation of the tight junction pathway and ZO-1 expression.


Assuntos
Anti-Inflamatórios/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Faecalibacterium prausnitzii/metabolismo , Mucosa Intestinal/metabolismo , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Células CACO-2 , Diabetes Mellitus Tipo 2/genética , Disbiose/genética , Disbiose/fisiopatologia , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/genética , Microbioma Gastrointestinal/fisiologia , Expressão Gênica/efeitos dos fármacos , Células HEK293 , Células HT29 , Humanos , Doenças Inflamatórias Intestinais/genética , Doenças Inflamatórias Intestinais/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Camundongos , Permeabilidade/efeitos dos fármacos , Proteínas de Junções Íntimas/genética , Junções Íntimas/efeitos dos fármacos
8.
Cytokine ; 121: 154718, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31153056

RESUMO

A single layer of epithelial cells creates an interface between the host and microorganisms colonizing the gastrointestinal tract. In a healthy intestine, commensal bacteria and their metabolites can interact with epithelial cells as they are identified by Toll-like receptors (TLRs); This interaction results in homeostasis and immune responses. The present study aimed at evaluating Faecalibacterium prausnitzii- and extracellular vesicles (EVs)-induced expression of involved genes in TLRs signaling pathway and cytokines production in Caco-2 cell line. In this study, Caco-2 cell line was treated with F. prausitzii and its EVs. Using the protein levels of 12 cytokines were also evaluated by ELISA assay. F. prausnitzii induced upregulation in FOS, JUN, TNF-α, NFKB1, TLR3, IKBKB and CD86 genes. Furthermore, stimulation of Caco-2 cells with EVs derived from F. prausnitzii induced upregulation of CXCL8, CCL2, FOS, MAP2K4, TLR7, TLR3, IRF1, NFKBIA and TNF-α genes. Based on ELISA assay, Caco-2 cells treated with F. prausnitzii and its EVs showed a significant increase in TNF-α, IL-4, IL-8, and IL-10 expression and significant decreased in IL-1, IL-2, IL-6, IL-12, IL-17a, IFN-γ compared to the control group (P < 0.05). In conclusion, EVs derived from F. prausnitzii showed greater efficacy in decreasing the inflammatory cytokines and increasing the anti-inflammatory cytokines, compared to F. prausnitzii. Our findings can be used as a theoretical model for EVs application in the potential treatment of inflammation.


Assuntos
Citocinas/genética , Enterócitos/metabolismo , Enterócitos/microbiologia , Vesículas Extracelulares/metabolismo , Faecalibacterium prausnitzii/metabolismo , Regulação da Expressão Gênica , Transdução de Sinais , Receptores Toll-Like/metabolismo , Células CACO-2 , Quimiocinas/genética , Quimiocinas/metabolismo , Análise por Conglomerados , Citocinas/metabolismo , Regulação para Baixo/genética , Humanos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Regulação para Cima/genética
9.
Mol Med Rep ; 20(1): 25-32, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31115531

RESUMO

Faecalibacterium prausnitzii is one of the most abundant bacteria in the human gut microbiota. This bacterium is reported to serve an important role in inflammatory bowel diseases. In the present study, the preventive effects of F. prausnitzii on a dextran sodium sulfate (DSS)­induced colitis model in mice were investigated. BALB/c mice were fed with 5% DSS in drinking water. Administration of live or inactivated F. prausnitzii was initiated 7 days prior to the start of DSS feeding. Mucosal cytokines were analyzed by reverse transcription­quantitative PCR. Histological analysis of colon mucosa was also performed. The symptoms of DSS­induced colitis (weight loss, diarrhea, bloody stools and colon shortening) were significantly improved in the group administered live F. prausnitzii, but not in the other groups. There were no significant differences in the expression of proinflammatory cytokines; however, the expression of mucosal cytokines appeared to be markedly reduced in the live F. prausnitzii­administered group compared with the DSS­fed control. The results suggested that preventive administration of 'live', but not inactivated, F. prausnitzii protected the colon against DSS­induced colitis. Live F. prausnitzii were also administered therapeutically following the induction of colitis, resulting in an improved histological score in mice.


Assuntos
Colite/microbiologia , Colite/terapia , Faecalibacterium prausnitzii/metabolismo , Microbioma Gastrointestinal/genética , Animais , Colite/induzido quimicamente , Colite/genética , Citocinas/genética , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Água Potável/administração & dosagem , Água Potável/microbiologia , Regulação da Expressão Gênica , Humanos , Mucosa Intestinal/microbiologia , Camundongos
10.
Psychoneuroendocrinology ; 104: 132-142, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30844607

RESUMO

The realization that the microbiota-gut-brain axis plays a critical role in health and disease,including neuropsychiatric disorders, is rapidly advancing.An abundance of preclinical studies have shown that psychobiotics acting via the brain-gut-axis can affect brain development, function and behavior. Here we tested whether potential psychobiotics Faecalibacterium prausnitzii (ATCC 27766) has anxiolytic and antidepressant-like effects and reverse the impact of chronic unpredictable mild stress (CUMS) in rats. The experiment was divided into two phases, the first stage was CUMS procedure period and the second stage was convalescence period. SD male rats were administered Faecalibacterium prausnitzii for 4 weeks prior to testing during each period. Behavior, growth status, SCFAs produced, plasma cytokine, endocrinology and bone mineral density (BMD) were assessed. Our findings indicate that the administration of F. prausnitzii had preventive and therapeutic effects on CUMS-induced depression-like and anxiety-like behavior. In addition, F. prausnitzii administration could significantly prevent the reduction of the whole-body, femur and tibia BMD during the recovery phase. Moreover, the growth status of rats fed the F. prausnitzii was better than the rats by CUMS. And F. prausnitzii administration led to higher levels of SCFAs in the cecum and higher levels of cytokines interleukin-10 (IL-10) in the plasma, prevented the effects on corticosterone, C-reaction protein and cytokines interleukin-6 (IL-6) release induced by CUMS, changes that were associated with the effects seen on behavior. These results provide further evidence that gut microflora play a role in anxiety and depression. Subject to the confirmation of these results, probiotics might offer a useful novel therapeutic approach to neuropathological disorders and/or as adjunct therapies in psychiatric disorders and support the recent broadening of the definition of psychobiotic. Finally, this study supports F. prausnitzii has significant potential as a psychobiotic.


Assuntos
Faecalibacterium prausnitzii/metabolismo , Probióticos/farmacologia , Estresse Psicológico/tratamento farmacológico , Animais , Ansiolíticos/farmacologia , Ansiedade/metabolismo , Transtornos de Ansiedade/metabolismo , Comportamento Animal/efeitos dos fármacos , Densidade Óssea/efeitos dos fármacos , Encéfalo/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Corticosterona/sangue , Citocinas/metabolismo , Depressão/metabolismo , Transtorno Depressivo/metabolismo , Modelos Animais de Doenças , Faecalibacterium prausnitzii/fisiologia , Masculino , Ratos , Ratos Sprague-Dawley , Estresse Psicológico/metabolismo
11.
Int Immunol ; 31(8): 499-514, 2019 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-30809639

RESUMO

Decreased levels of Faecalibacterium prausnitzii (F. prausnitzii), whose supernatant plays an anti-inflammatory effect, are frequently found in inflammatory bowel disease (IBD) patients. However, the anti-inflammatory products in F. prausnitzii supernatant and the mechanism have not been fully investigated. Here we found that F. prausnitzii and F. prausnitzii-derived butyrate were decreased in the intestines of IBD patients. Supplementation with F. prausnitzii supernatant and butyrate could ameliorate colitis in an animal model. Butyrate, but not other substances produced by F. prausnitzii, exerted an anti-inflammatory effect by inhibiting the differentiation of T helper 17 (Th17) cells. The mechanism underlying the anti-inflammatory effects of the butyrate produced by F. prausnitzii involved the enhancement of the acetylation-promoted degradation of c-Myc through histone deacetylase 3 (HDAC3) inhibition. In conclusion, F. prausnitzii produced butyrate to decrease Th17 differentiation and attenuate colitis through inhibiting HDAC3 and c-Myc-related metabolism in T cells. The use of F. prausnitzii may be an effective new approach to decrease the level of Th17 cells in the treatment of inflammatory diseases.


Assuntos
Anti-Inflamatórios/farmacologia , Butiratos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Faecalibacterium prausnitzii/metabolismo , Histona Desacetilases/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Células Th17/efeitos dos fármacos , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/metabolismo , Butiratos/química , Butiratos/metabolismo , Colite/induzido quimicamente , Colite/metabolismo , Colite/patologia , Faecalibacterium prausnitzii/química , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ratos , Ratos Sprague-Dawley , Células Th17/citologia , Células Th17/metabolismo , Ácido Trinitrobenzenossulfônico/administração & dosagem
12.
Drug Metab Dispos ; 47(3): 194-202, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30598508

RESUMO

Tacrolimus exhibits low and variable drug exposure after oral dosing, but the contributing factors remain unclear. Based on our recent report showing a positive correlation between fecal abundance of Faecalibacterium prausnitzii and oral tacrolimus dose in kidney transplant patients, we tested whether F. prausnitzii and other gut abundant bacteria are capable of metabolizing tacrolimus. Incubation of F. prausnitzii with tacrolimus led to production of two compounds (the major one named M1), which was not observed upon tacrolimus incubation with hepatic microsomes. Isolation, purification, and structure elucidation using mass spectrometry and nuclear magnetic resonance spectroscopy indicated that M1 is a C-9 keto-reduction product of tacrolimus. Pharmacological activity testing using human peripheral blood mononuclear cells demonstrated that M1 is 15-fold less potent than tacrolimus as an immunosuppressant. Screening of 22 gut bacteria species revealed that most Clostridiales bacteria are extensive tacrolimus metabolizers. Tacrolimus conversion to M1 was verified in fresh stool samples from two healthy adults. M1 was also detected in the stool samples from kidney transplant recipients who had been taking tacrolimus orally. Together, this study presents gut bacteria metabolism as a previously unrecognized elimination route of tacrolimus, potentially contributing to the low and variable tacrolimus exposure after oral dosing.


Assuntos
Faecalibacterium prausnitzii/metabolismo , Microbioma Gastrointestinal/fisiologia , Imunossupressores/metabolismo , Tacrolimo/metabolismo , Administração Oral , Adulto , Idoso , Células Cultivadas , Relação Dose-Resposta a Droga , Fezes/química , Feminino , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/prevenção & controle , Voluntários Saudáveis , Humanos , Imunossupressão/métodos , Imunossupressores/administração & dosagem , Imunossupressores/análise , Transplante de Rim/efeitos adversos , Leucócitos Mononucleares/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Simbiose , Tacrolimo/administração & dosagem , Tacrolimo/análise
13.
Artigo em Inglês | MEDLINE | ID: mdl-32083024

RESUMO

Migraine is a very common, multifactorial, and recurrent central nervous system disorder that causes throbbing headache, photophobia, phonophobia, nausea, and disability. Migraine occurs more often in females, and its complex physiopathology is not yet fully understood. An increasing number of gastrointestinal disorders have been linked to the occurrence of migraine suggesting that gut microbiota might play a pivotal role in migraine through the gut-brain axis. In the present work, we performed a metagenome-wide association study (MWAS) to determine the relationship between gut microbiota and migraine by analyzing 108 shotgun-sequenced fecal samples obtained from elderly women who suffer from migraine and matched healthy controls. Notably, the alpha diversity was significantly decreased in the migraine group at species, genus, and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthologous levels. Firmicutes, especially the "unfriendly" Clostridium spp., were significantly enriched in the migraine group. Conversely, the healthy controls held more beneficial microorganisms, such as Faecalibacterium prausnitzii, Bifidobacterium adolescentis, and Methanobrevibacter smithii. For functional modules, the migraine group was enriched in gut-brain modules (GBMs) including kynurenine degradation and γ-aminobutyric acid (GABA) synthesis. However, the healthy controls held higher gut metabolic modules (GMMs) including glycolysis, homoacetogenesis, and GBMs including quinolinic acid degradation and S-adenosyl methionine (SAM) synthesis. The differences in gut microbiota composition and function between the migraine and healthy groups provided new information as well as novel therapeutic targets and strategies for migraine treatment, which could help to improve the early diagnosis of the disease, as well as the long-term prognosis and the life quality of patients suffering from migraine.


Assuntos
Bactérias/metabolismo , Microbioma Gastrointestinal/fisiologia , Trato Gastrointestinal/microbiologia , Transtornos de Enxaqueca/terapia , Adenosina/análogos & derivados , Adenosina/metabolismo , Idoso , Bactérias/classificação , Bifidobacterium adolescentis/metabolismo , Faecalibacterium prausnitzii/metabolismo , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/genética , Humanos , Metagenoma , Methanobrevibacter/metabolismo , Prognóstico , Ácido Quinolínico/metabolismo , Espermidina/análogos & derivados , Espermidina/metabolismo
14.
BMC Genomics ; 19(1): 931, 2018 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-30547746

RESUMO

BACKGROUND: Faecalibacterium prausnitzii is a ubiquitous member of the human gut microbiome, constituting up to 15% of the total bacteria in the human gut. Substantial evidence connects decreased levels of F. prausnitzii with the onset and progression of certain forms of inflammatory bowel disease, which has been attributed to its anti-inflammatory potential. Two phylogroups of F. prausnitzii have been identified, with a decrease in phylogroup I being a more sensitive marker of intestinal inflammation. Much of the genomic and physiological data available to date was collected using phylogroup II strains. Little analysis of F. prausnitzii genomes has been performed so far and genetic differences between phylogroups I and II are poorly understood. RESULTS: In this study we sequenced 11 additional F. prausnitzii genomes and performed comparative genomics to investigate intraspecies diversity, functional gene complement and the mobilome of 31 high-quality draft and complete genomes. We reveal a very low level of average nucleotide identity among F. prausnitzii genomes and a high level of genome plasticity. Two genomogroups can be separated based on differences in functional gene complement, albeit that this division does not fully agree with separation based on conserved gene phylogeny, highlighting the importance of horizontal gene transfer in shaping F. prausnitzii genomes. The difference between the two genomogroups is mainly in the complement of genes associated with catabolism of carbohydrates (such as a predicted sialidase gene in genomogroup I) and amino acids, as well as defense mechanisms. CONCLUSIONS: Based on the combination of ANI of genomic sequences, phylogenetic analysis of core proteomes and functional differences we propose to separate the species F. prausnitzii into two new species level taxa: F. prausnitzii sensu stricto (neotype strain A2-165T = DSM 17677T = JCM 31915T) and F. moorei sp. nov. (type strain ATCC 27768T = NCIMB 13872T).


Assuntos
Faecalibacterium prausnitzii/genética , Genoma Bacteriano , Análise por Conglomerados , Hibridização Genômica Comparativa , Faecalibacterium prausnitzii/classificação , Faecalibacterium prausnitzii/metabolismo , Fezes/microbiologia , Microbioma Gastrointestinal , Humanos , Filogenia , Análise de Componente Principal , Proteoma , RNA Ribossômico 16S/química , RNA Ribossômico 16S/isolamento & purificação , RNA Ribossômico 16S/metabolismo , Análise de Sequência de DNA , Especificidade da Espécie
15.
Inflamm Bowel Dis ; 24(9): 1926-1940, 2018 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-29796620

RESUMO

BACKGROUND: Inflammatory bowel disease (IBD)-associated dysbiosis is characterized by a loss of Faecalibacterium prausnitzii, whose supernatant exerts an anti-inflammatory effect. However, the anti-inflammatory substances in F. prausnitzii supernatant and the mechanism in ameliorating colitis in IBD have not yet been fully investigated. METHODS: Experimental colitis models were induced and evaluated by clinical examination and histopathology. Levels of cytokines and ratio of T cells were detected by enzyme-linked immunosorbent assay and flow cytometry analysis, respectively. F. prausnitzii supernatant was separated by macroporous resins. After extraction, the substances in supernatant were identified by gas chromatography-mass spectrometer. T-cell differentiation assay was conducted in vitro. Changes in signaling pathways were examined by immunoblot, immunohistochemistry, and immunofluorescent staining. RESULTS: We found that the supernatant of F. prausnitzii could regulate T helper 17 cell (Th17)/regulatory T cell (Treg) differentiation. Then, we identified butyrate produced by F. prausnitzii that played the anti-inflammatory effects by inhibiting interleukin (IL)-6/signal transducer and the activator of transcription 3 (STAT3)/IL-17 pathway and promoting forkhead box protein P3 (Foxp3). Finally, we demonstrated that the target of butyrate was histone deacetylase 1 (HDAC1). CONCLUSIONS: It is butyrate, instead of other substances produced by F. prausnitzii, that maintains Th17/Treg balance and exerts significant anti-inflammatory effects in colorectal colitis rodents, by inhibiting HDAC1 to promote Foxp3 and block the IL-6/STAT3/IL-17 downstream pathway. F. prausnitzii could be an option for further investigation for IBD treatment. Targeting the butyrate-HDAC1-T-cell axis offers an effective novel approach in the treatment of inflammatory disease.


Assuntos
Butiratos/metabolismo , Colite/microbiologia , Faecalibacterium prausnitzii/metabolismo , Histona Desacetilase 1/metabolismo , Linfócitos T Reguladores/microbiologia , Células Th17/microbiologia , Animais , Diferenciação Celular , Colite/induzido quimicamente , Colo/microbiologia , Modelos Animais de Doenças , Disbiose/microbiologia , Histona Desacetilase 1/antagonistas & inibidores , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ratos , Ratos Sprague-Dawley , Reto/microbiologia , Transdução de Sinais
16.
JPEN J Parenter Enteral Nutr ; 42(7): 1156-1167, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29385239

RESUMO

BACKGROUND: Clostridium difficile (CD) infection (CDI) increases patient morbidity, mortality and healthcare costs. Antibiotic treatment induces gut dysbiosis and is both a major risk factor for CD colonization and treatment of CDI. Probiotics have been trialed to support commensal gut microbiota and reduce CDI. This study investigated commensal microbe Faecalibacterium prausnitzii (FP) and a prebiotic, both known to yield butyrate and be anti-inflammatory and immunomodulatory, on CD colonization and gut integrity in mice. METHODS: Mice were randomly grouped and supplemented daily with FP, prebiotic, FP + prebiotic, FP/prebiotic supernatant, or saline throughout the entire study. Following treatment with clindamycin for 3 days, mice were exposed to CD. Feces were collected at baseline, the day after antibiotic, and 1, 3, and 5 days after CD exposure and cultured for bacterial overgrowth and CD colonization. On days 1 and 5 after CD exposure, mice were randomly euthanized, and proximal colon was dissected for histological analysis and preparation of RNA for analysis of proinflammatory and anti-inflammatory cytokines. RESULTS: Although all mice exhibited bacterial overgrowth and CD colonization, bacterial burden resolved quicker in the FP + prebiotic group. This was associated with induction and resolution of innate immune responses, anion exchanger, and tight junction protein preservation in proximal colon. CD toxin virulence potential was questionable as expression of CD toxin B receptor was depleted in the FP + prebiotic group. CONCLUSION: Supplementation with anti-inflammatory butyrate-supporting commensal bacteria and prebiotic may support innate immune responses and minimize bacterial burden and negative effects during antibiotic and CD exposure.


Assuntos
Antibacterianos/efeitos adversos , Clostridioides difficile/crescimento & desenvolvimento , Infecções por Clostridium/tratamento farmacológico , Faecalibacterium prausnitzii , Microbioma Gastrointestinal , Prebióticos , Probióticos/uso terapêutico , Animais , Proteínas de Transporte de Ânions/metabolismo , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Butiratos/metabolismo , Butiratos/farmacologia , Clindamicina/efeitos adversos , Clostridioides difficile/efeitos dos fármacos , Clostridioides difficile/metabolismo , Clostridioides difficile/patogenicidade , Infecções por Clostridium/imunologia , Infecções por Clostridium/metabolismo , Infecções por Clostridium/microbiologia , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/microbiologia , Citocinas/metabolismo , Modelos Animais de Doenças , Disbiose/etiologia , Faecalibacterium prausnitzii/crescimento & desenvolvimento , Faecalibacterium prausnitzii/metabolismo , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Ratos Sprague-Dawley , Receptores Imunológicos/metabolismo , Solanum tuberosum/química , Amido/farmacologia , Amido/uso terapêutico , Proteínas de Junções Íntimas/metabolismo
17.
Biochemistry ; 56(45): 6006-6014, 2017 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-29045794

RESUMO

Self-cleaving ribozymes were discovered 30 years ago and have been found throughout nature, from bacteria to animals, but little is known about their biological functions and regulation, particularly how cofactors and metabolites alter their activity. A hepatitis delta virus-like self-cleaving ribozyme maps upstream of a phosphoglucosamine mutase (glmM) open reading frame in the genome of the human gut bacterium Faecalibacterium prausnitzii. The presence of a ribozyme in the untranslated region of glmM suggests a regulation mechanism of gene expression. In the bacterial hexosamine biosynthesis pathway, the enzyme glmM catalyzes the isomerization of glucosamine 6-phosphate into glucosamine 1-phosphate. In this study, we investigated the effect of these metabolites on the co-transcriptional self-cleavage rate of the ribozyme. Our results suggest that glucosamine 6-phosphate, but not glucosamine 1-phosphate, is an allosteric ligand that increases the self-cleavage rate of drz-Fpra-1, providing the first known example of allosteric modulation of a self-cleaving ribozyme by the substrate of the adjacent gene product. Given that the ribozyme is activated by the glmM substrate, but not the product, this allosteric modulation may represent a potential feed-forward mechanism of gene expression regulation in bacteria.


Assuntos
Faecalibacterium prausnitzii/enzimologia , Faecalibacterium prausnitzii/genética , Regulação Enzimológica da Expressão Gênica , Fosfoglucomutase/metabolismo , RNA Catalítico/metabolismo , Regulação Alostérica , Sequência de Bases , Faecalibacterium prausnitzii/metabolismo , Genoma Bacteriano , Glucosamina/análogos & derivados , Glucosamina/metabolismo , Glucose-6-Fosfato/análogos & derivados , Glucose-6-Fosfato/metabolismo , Vírus Delta da Hepatite/enzimologia , Conformação de Ácido Nucleico , Fosfoglucomutase/genética , RNA Catalítico/genética
18.
Benef Microbes ; 8(3): 473-490, 2017 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-28548573

RESUMO

Four selected butyrate-producing colon bacterial strains belonging to Clostridium cluster IV (Butyricicoccus pullicaecorum DSM 23266T and Faecalibacterium prausnitzii DSM 17677T) and XIVa (Eubacterium hallii DSM 17630 and Eubacterium rectale CIP 105953T) were studied as to their capacity to degrade inulin-type fructans and concomitant metabolite production. Cultivation of these strains was performed in bottles and fermentors containing a modified medium for colon bacteria, including acetate, supplemented with either fructose, oligofructose, or inulin as the sole energy source. Inulin-type fructan degradation was not a general characteristic among these strains. B. pullicaecorum DSM 23266T and E. hallii DSM 17630 could only ferment fructose and did not degrade oligofructose or inulin. E. rectale CIP 105953T and F. prausnitzii DSM 17677T fermented fructose and could degrade both oligofructose and inulin. All chain length fractions of oligofructose were degraded simultaneously (both strains) and both long and short chain length fractions of inulin were degraded either simultaneously (E. rectale CIP 105953T) or consecutively (F. prausnitzii DSM 17677T), indicating an extracellular polymer degradation mechanism. B. pullicaecorum DSM 23266T and E. hallii DSM 17630 produced high concentrations of butyrate, CO2, and H2 from fructose. E. rectale CIP 105953T produced lactate, butyrate, CO2, and H2, from fructose, oligofructose, and inulin, whereas F. prausnitzii DSM 17677T produced butyrate, formate, CO2, and traces of lactate from fructose, oligofructose, and inulin. Based on carbon recovery and theoretical metabolite production calculations, an adapted stoichiometrically balanced metabolic pathway for butyrate, formate, lactate, CO2, and H2 production by members of both Clostridium cluster IV and XIVa butyrate-producing bacteria was constructed.


Assuntos
Eubacterium/metabolismo , Faecalibacterium prausnitzii/metabolismo , Frutose/metabolismo , Inulina/metabolismo , Oligossacarídeos/metabolismo , Butiratos/metabolismo , Dióxido de Carbono/metabolismo , Colo/metabolismo , Colo/microbiologia , Fermentação/fisiologia , Formiatos/metabolismo , Humanos , Hidrogênio/metabolismo , Ácido Láctico/metabolismo
19.
Antonie Van Leeuwenhoek ; 109(10): 1389-96, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27431681

RESUMO

The human gut microbiota plays an important role in human health and might also be implicated in kidney disease. The interest in butyrate producing bacteria has recently increased and is a poorly understood faecal condition in chronic kidney disease (CKD). Therefore, we evaluated differences of the butyrate producing species Roseburia spp. and Faecalibacterium prausnitzii in the faeces of Chinese patients with CKD. A case-control study was carried out for 65 CKD patients and 20 healthy controls. Differences were quantitatively validated using quantitative real-time polymerase chain reaction (qPCR). Spearman rank correlation was used to analyse the correlation between gut microbiota and clinical variables. Roseburia spp. and F. prausnitzii were significantly different in CKD patients and controls (p = 0.001; p = 0.025, respectively) and reduced more markedly in end stage renal disease (p = 0.000; p = 0.003, respectively) and microinflammation (p = 0.004; p = 0.001, respectively). Roseburia spp. and F. prausnitzii were negatively associated with C-reactive protein in plasma (r = -0.493, p = 0.00; r = -0.528, p = 0.000; respectively) and Cystatin C (r = -0.321, p = 0.006; r = -0.445, p = 0.000; respectively). They were positively associated with eGFR (r = 0.347, p = 0.002; r = 0.416, p = 0.000; respectively). The negative correlation between Roseburia spp., F. prausnitzii and CRP and renal function suggested that the depletion of butyrate producing bacteria may contribute to CKD-associated inflammation and CKD progression. Roseburia spp. and F. prausnitzii may thus serve as 'microbiomarkers'.


Assuntos
Butiratos/metabolismo , Clostridiales/metabolismo , Faecalibacterium prausnitzii/metabolismo , Insuficiência Renal Crônica/microbiologia , Adulto , Estudos de Casos e Controles , Progressão da Doença , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal , Humanos , Masculino , Reação em Cadeia da Polimerase em Tempo Real
20.
World J Gastroenterol ; 22(22): 5201-10, 2016 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-27298563

RESUMO

AIM: To explore the preventive and therapeutic effects of Faecalibacterium prausnitzii (F. prausnitzii) supernatant on dextran sulfate sodium (DSS) induced colitis in mice. METHODS: Forty C57BL/6J male mice were randomly divided into four groups: control group, model group, treatment group, and prevention group. Mice were weighed daily. On day 10, the colon length was measured, the colorectal histopathologic damage score (HDS) was assessed, and plasma interleukin (IL)-17A, IL-6, and IL-4 levels were detected by enzyme-linked immunosorbent assay. The expression of transcription factor retinoic acid-related orphan receptor-γt (RORγt) and IL-17A in colon inflammatory mucosa tissue were determined by immunohistochemical assay, and the expression levels of RORγt mRNA, IL-17A mRNA, and IL-6 mRNA were detected by real-time quantitative polymerase chain reaction (PCR). The proportion of Th17 in mononuclear cells in spleen was assayed by fluorescence activated cell sorter. RESULTS: When compared with the model group, the colon length (P < 0.05) and body weight (P < 0.01) in the treatment and prevention groups were significantly increased, and the colon HDS was decreased (P < 0.05 and P < 0.01). There was no statistical difference between the treatment group and prevention group. After treatment with F. prausnitzii supernatant, the plasma levels of IL-17A and IL-6 (P < 0.05), the protein and mRNA expression of IL-17A and RORγt, and the Th17 cell ratio of spleen cells (P < 0.01) were significantly decreased compared to the model group. Plasma IL-4 level in the prevention group was significantly higher than that in the model group (P < 0.05), but there was no significant difference between these two groups in the expression of IL-6 in both the plasma and colon mucosa tissues. CONCLUSION: F. prausnitzii supernatant exerts protective and therapeutic effects on DSS-induced colitis in mice, probably via inhibition of Th17 differentiation and IL-17A secretion in the plasma and colon mucosa tissues. It can also improve colitis in mice by downregulating IL-6 and prevent colitis by upregulating IL-4.


Assuntos
Anti-Inflamatórios/farmacologia , Colite/prevenção & controle , Colo/efeitos dos fármacos , Suplementos Nutricionais , Faecalibacterium prausnitzii/metabolismo , Fármacos Gastrointestinais/farmacologia , Células Th17/efeitos dos fármacos , Animais , Colite/sangue , Colite/induzido quimicamente , Colite/imunologia , Colo/imunologia , Colo/metabolismo , Colo/patologia , Sulfato de Dextrana , Mediadores da Inflamação/sangue , Interleucina-17/sangue , Interleucina-17/genética , Interleucina-4/sangue , Interleucina-6/sangue , Interleucina-6/genética , Masculino , Camundongos Endogâmicos C57BL , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Células Th17/imunologia , Células Th17/metabolismo , Fatores de Tempo
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