RESUMO
Gut microbiota alterations have been described in several diseases with altered gastrointestinal (GI) motility, and awareness is increasing regarding the role of the gut microbiome in modulating GI function. Serotonin [5-hydroxytryptamine (5-HT)] is a key regulator of GI motility and secretion. To determine the relationship among gut microbes, colonic contractility, and host serotonergic gene expression, we evaluated mice that were germ-free (GF) or humanized (HM; ex-GF colonized with human gut microbiota). 5-HT reduced contractile duration in both GF and HM colons. Microbiota from HM and conventionally raised (CR) mice significantly increased colonic mRNAs Tph1 [(tryptophan hydroxylase) 1, rate limiting for mucosal 5-HT synthesis; P < 0.01] and chromogranin A (neuroendocrine secretion; P < 0.01), with no effect on monoamine oxidase A (serotonin catabolism), serotonin receptor 5-HT4, or mouse serotonin transporter. HM and CR mice also had increased colonic Tph1 protein (P < 0.05) and 5-HT concentrations (GF, 17 ± 3 ng/mg; HM, 25 ± 2 ng/mg; and CR, 35 ± 3 ng/mg; P < 0.05). Enterochromaffin (EC) cell numbers (cells producing 5-HT) were unchanged. Short-chain fatty acids (SCFAs) promoted TPH1 transcription in BON cells (human EC cell model). Thus, gut microbiota acting through SCFAs are important determinants of enteric 5-HT production and homeostasis.
Assuntos
Colo/metabolismo , Sistema Digestório/microbiologia , Células Enterocromafins/metabolismo , Ácidos Graxos Voláteis/metabolismo , Serotonina/biossíntese , Animais , Contagem de Células , Linhagem Celular , Cromogranina A/genética , Colo/citologia , Colo/microbiologia , Células Enterocromafins/citologia , Células Enterocromafins/microbiologia , Feminino , Motilidade Gastrointestinal , Vida Livre de Germes , Humanos , Masculino , Camundongos , Microbiota , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais , Triptofano Hidroxilase/genética , Triptofano Hidroxilase/metabolismoRESUMO
BACKGROUND: Obesity is associated with accumulation of macrophages in white adipose tissue (WAT), which contribute to the development of insulin resistance. Germ-free (GF) mice have reduced adiposity and are protected against diet-induced obesity, OBJECTIVE: To investigate whether the gut microbiota and, specifically, gut-derived lipopolysaccharide (LPS) promote WAT inflammation and contribute to impaired glucose metabolism. METHOD: Macrophage composition and expression of proinflammatory and anti-inflammatory markers were compared in WAT of GF, conventionally raised and Escherichia coli-monocolonised mice. Additionally, glucose and insulin tolerance in these mice was determined. RESULTS: The presence of a gut microbiota resulted in impaired glucose metabolism and increased macrophage accumulation and polarisation towards the proinflammatory M1 phenotype in WAT. Monocolonisation of GF mice for 4 weeks with E. coli W3110 or the isogenic strain MLK1067 (which expresses LPS with reduced immunogenicity) resulted in impaired glucose and insulin tolerance and promoted M1 polarisation of CD11b cells in WAT. However, colonisation with E. coli W3110 but not MLK1067 promoted macrophage accumulation and upregulation of proinflammatory and anti-inflammatory gene expression as well as JNK phosphorylation. CONCLUSION: Gut microbiota induced LPS-dependent macrophage accumulation in WAT, whereas impairment of systemic glucose metabolism was not dependent on LPS. These results indicate that macrophage accumulation in WAT does not always correlate with impaired glucose metabolism.
Assuntos
Tecido Adiposo Branco/metabolismo , Escherichia coli/metabolismo , Intolerância à Glucose/microbiologia , Resistência à Insulina , Intestinos/microbiologia , Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Tecido Adiposo Branco/imunologia , Tecido Adiposo Branco/patologia , Animais , Biomarcadores/metabolismo , Citometria de Fluxo , Vida Livre de Germes , Intolerância à Glucose/imunologia , Intolerância à Glucose/metabolismo , Intolerância à Glucose/patologia , Immunoblotting , Imuno-Histoquímica , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/microbiologia , Inflamação/patologia , Masculino , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Obesity is characterised by altered gut microbiota, low-grade inflammation and increased endocannabinoid (eCB) system tone; however, a clear connection between gut microbiota and eCB signalling has yet to be confirmed. Here, we report that gut microbiota modulate the intestinal eCB system tone, which in turn regulates gut permeability and plasma lipopolysaccharide (LPS) levels. The impact of the increased plasma LPS levels and eCB system tone found in obesity on adipose tissue metabolism (e.g. differentiation and lipogenesis) remains unknown. By interfering with the eCB system using CB(1) agonist and antagonist in lean and obese mouse models, we found that the eCB system controls gut permeability and adipogenesis. We also show that LPS acts as a master switch to control adipose tissue metabolism both in vivo and ex vivo by blocking cannabinoid-driven adipogenesis. These data indicate that gut microbiota determine adipose tissue physiology through LPS-eCB system regulatory loops and may have critical functions in adipose tissue plasticity during obesity.
Assuntos
Adipogenia , Tecido Adiposo/metabolismo , Translocação Bacteriana , Moduladores de Receptores de Canabinoides/metabolismo , Endocanabinoides , Intestinos/microbiologia , Lipopolissacarídeos/sangue , Obesidade/metabolismo , Receptor CB1 de Canabinoide/metabolismo , Adipogenia/efeitos dos fármacos , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/fisiopatologia , Animais , Ácidos Araquidônicos/metabolismo , Translocação Bacteriana/efeitos dos fármacos , Células CACO-2 , Modelos Animais de Doenças , Dronabinol/análogos & derivados , Dronabinol/farmacologia , Glicerídeos/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fator 88 de Diferenciação Mieloide/deficiência , Fator 88 de Diferenciação Mieloide/genética , Obesidade/microbiologia , Obesidade/fisiopatologia , Permeabilidade , Piperidinas/farmacologia , Alcamidas Poli-Insaturadas/metabolismo , Prebióticos , Pirazóis/farmacologia , RNA Mensageiro/metabolismo , Receptor CB1 de Canabinoide/efeitos dos fármacos , Receptor CB1 de Canabinoide/genética , RimonabantoRESUMO
The gut microbiota has recently been identified as an environmental factor that may promote metabolic diseases. To investigate the effect of gut microbiota on host energy and lipid metabolism, we compared the serum metabolome and the lipidomes of serum, adipose tissue, and liver of conventionally raised (CONV-R) and germ-free mice. The serum metabolome of CONV-R mice was characterized by increased levels of energy metabolites, e.g., pyruvic acid, citric acid, fumaric acid, and malic acid, while levels of cholesterol and fatty acids were reduced. We also showed that the microbiota modified a number of lipid species in the serum, adipose tissue, and liver, with its greatest effect on triglyceride and phosphatidylcholine species. Triglyceride levels were lower in serum but higher in adipose tissue and liver of CONV-R mice, consistent with increased lipid clearance. Our findings show that the gut microbiota affects both host energy and lipid metabolism and highlights its role in the development of metabolic diseases.
Assuntos
Metabolismo Energético , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/microbiologia , Metabolismo dos Lipídeos , Metagenoma/fisiologia , Adipócitos Brancos/metabolismo , Animais , Quilomícrons/sangue , Vida Livre de Germes , Absorção Intestinal , Fígado/metabolismo , Masculino , Espectrometria de Massas , Metaboloma , Camundongos , Fosfatidilcolinas/sangue , Fosfatidilcolinas/metabolismo , Ácido Pirúvico/sangue , Ácido Pirúvico/metabolismo , Ácidos Tricarboxílicos/sangue , Ácidos Tricarboxílicos/metabolismo , Triglicerídeos/sangue , Triglicerídeos/metabolismoRESUMO
Obesity is a worldwide epidemic, threatening both industrialized and developing countries, and is accompanied by a dramatic increase in obesity-related disorders, including type 2 diabetes mellitus, hypertension, cardiovascular diseases, and nonalcoholic fatty liver disease. Recent studies have shown that the gut microbial community (microbiota) is an environmental factor that regulates obesity by increasing energy harvest from the diet and by regulating peripheral metabolism. However, there are no data on how obesogenic microbiotas are established and whether this process is determined during infancy. The sterile fetus is born into a microbial world and is immediately colonized by numerous species originating from the surrounding ecosystems, especially the maternal vaginal and fecal microflora. This initial microbiota develops into a complex ecosystem in a predictable fashion determined by internal (eg, oxygen depletion) and external (eg, mode of birth, impact of environment, diet, hospitalization, application of antibiotics) factors. We discuss how the gut microbiota regulates obesity and how environmental factors that affect the establishment of the gut microbiota during infancy may contribute to obesity later in life.
Assuntos
Intestinos/microbiologia , Obesidade/microbiologia , Animais , Humanos , Imunidade Inata/fisiologia , Lactente , Intestinos/imunologia , Redes e Vias Metabólicas , Camundongos , Obesidade/imunologia , Obesidade/metabolismoRESUMO
BACKGROUND: Microbial constituents of the gut microbiome interact with each other and the host to alter the luminal environment and impact development, motility, and homeostasis of the gut. Powerful methods are becoming available to investigate connections between the gut microbiome and human health. While high-throughput sequencing of 16S rRNA genes can be used to identify and enumerate microbes in the gut, advances in several techniques (e.g., metagenomics, metatranscriptomics, metabolomics, and metaproteomics) are providing a clearer view as to the specific activities of the microbiota in the context of functional host-microbe interactions. Testing emergent hypotheses regarding microbial effects on host biology, which arise from analyses of 'Big Data' generated from massive parallel high-throughput sequencing technology and spectroscopic techniques, to guide translational research is an important goal for the future. Insights regarding the fundamental operating principles of the gut microbiota should lay the foundation for rational manipulation of the microbiota to promote human health. PURPOSE: In this review, we provide an overview of current research on the gut microbiome emphasizing current state-of-the-art technologies, approaches, and directions for improvement of our understanding of the impact of the gut microbiota with specific focus on gastrointestinal motility disorders.
Assuntos
Gastroenteropatias/microbiologia , Trato Gastrointestinal/microbiologia , Animais , Humanos , MetagenomaRESUMO
Recently, we demonstrated that colonic and adipose expression of SAA3 was modulated by the gut microbiota and Toll-like receptor signaling in mice. We observed that SAA3 was expressed by colonic epithelial cells and that its expression was induced in a murine colonocyte cell line following lipopolysaccharide stimulation and nuclear NFκB translocation. In this addendum, we extend this initial study and suggest that SAA3 (1) resembles human SAA1 both in amino acid homology and tissue distribution, (2) appears to have autocrine or paracrine effects rather than endocrine, and (3) binds to bacteria within the gastrointestinal tract. Although speculative, these observations raise the possibility that SAA3 may promote local inflammation in adipose tissue that affects insulin signaling and also function as an antimicrobial agent in the colon.
RESUMO
The gut microbiota has been proposed as an environmental factor that affects the development of metabolic and inflammatory diseases in mammals. Recent reports indicate that gut bacteria-derived lipopolysaccharide (LPS) can initiate obesity and insulin resistance in mice; however, the molecular interactions responsible for microbial regulation of host metabolism and mediators of inflammation have not been studied in detail. Hepatic serum amyloid A (SAA) proteins are markers and proposed mediators of inflammation that exhibit increased levels in serum of insulin-resistant mice. Adipose tissue-derived SAA3 displays monocyte chemotactic activity and may play a role in metabolic inflammation associated with obesity and insulin resistance. To investigate a potential mechanistic link between the intestinal microbiota and induction of proinflammatory host factors, we performed molecular analyses of germ-free, conventionally raised and genetically modified Myd88-/- mouse models. SAA3 expression was determined to be significantly augmented in adipose (9.9+/-1.9-fold; P<0.001) and colonic tissue (7.0+/-2.3-fold; P<0.05) by the presence of intestinal microbes. In the colon, we provided evidence that SAA3 is partially regulated through the Toll-like receptor (TLR)/MyD88/NF-kappaB signaling axis. We identified epithelial cells and macrophages as cellular sources of SAA3 in the colon and found that colonic epithelial expression of SAA3 may be part of an NF-kappaB-dependent response to LPS from gut bacteria. In vitro experiments showed that LPS treatments of both epithelial cells and macrophages induced SAA3 expression (27.1+/-2.5-fold vs. 1.6+/-0.1-fold, respectively). Our data suggest that LPS, and potentially other products of the indigenous gut microbiota, might elevate cytokine expression in tissues and thus exacerbate chronic low-grade inflammation observed in obesity.
Assuntos
Tecido Adiposo/metabolismo , Colo/metabolismo , Epitélio/metabolismo , Regulação da Expressão Gênica , Intestinos/microbiologia , Proteína Amiloide A Sérica/biossíntese , Animais , Variação Genética , Inflamação , Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Masculino , Camundongos , Obesidade/metabolismo , Transdução de Sinais , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Uropathogenic Escherichia coli (UPEC), the principal cause of urinary tract infection in women, colonizes the gut as well as the genitourinary tract. Studies of mice inoculated with UTI89, a sequenced isolate, have revealed a complex life cycle that includes formation of intracellular bacterial communities (IBCs) in bladder urothelial cells. To understand how UPEC adapts to life in IBCs, we have used GeneChips and/or quantitative reverse transcriptase PCR to study UTI89 recovered from the distal gut of gnotobiotic mice and from IBCs harvested by laser capture microdissection from the bladder urothelium of infected C3H/HeJ female mice. Host responses were characterized in laser capture microdissected urothelial cells that do or do not contain IBCs. The results reveal components of ferric iron acquisition systems in UTI89 that are expressed at significantly higher levels in IBCs compared with the intestine, including the hemin receptor chuA (1,390 +/- 188-fold). Localized urothelial responses to IBCs help oppose bacterial salvage of host cell iron (e.g. up-regulation of Tfrc (transferrin receptor) and Lcn2 (lipocalin 2)), facilitate glucose import (e.g. Hk2 (hexokinase 2)), and maintain epithelial structural integrity (e.g. Ivl (involucrin) and Sbsn (suprabasin)). DeltachuA mutants produce significantly smaller IBCs compared with wild type UTI89. This difference was not observed in strains lacking sitA (ABC-type iron/manganese transporter subunit), iroN (salmochelin receptor), hlyA (alpha-hemolysin), or entF (enterobactin synthetase subunit). Together, these studies indicate that heme- and siderophore-associated iron play key roles in IBC development and provide a series of microbial and host biomarkers for comparing UPEC strains isolated from humans.
Assuntos
Infecções por Escherichia coli/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Urotélio/metabolismo , Urotélio/microbiologia , Animais , Transporte Biológico , Biomarcadores/química , Biotinilação , Infecções por Escherichia coli/genética , Feminino , Genoma Bacteriano , Glucose/metabolismo , Humanos , Intestinos/microbiologia , Camundongos , Camundongos Endogâmicos C3H , Regulação para CimaRESUMO
Escherichia coli is a model laboratory bacterium, a species that is widely distributed in the environment, as well as a mutualist and pathogen in its human hosts. As such, E. coli represents an attractive organism to study how environment impacts microbial genome structure and function. Uropathogenic E. coli (UPEC) must adapt to life in several microbial communities in the human body, and has a complex life cycle in the bladder when it causes acute or recurrent urinary tract infection (UTI). Several studies designed to identify virulence factors have focused on genes that are uniquely represented in UPEC strains, whereas the role of genes that are common to all E. coli has received much less attention. Here we describe the complete 5,065,741-bp genome sequence of a UPEC strain recovered from a patient with an acute bladder infection and compare it with six other finished E. coli genome sequences. We searched 3,470 ortholog sets for genes that are under positive selection only in UPEC strains. Our maximum likelihood-based analysis yielded 29 genes involved in various aspects of cell surface structure, DNA metabolism, nutrient acquisition, and UTI. These results were validated by resequencing a subset of the 29 genes in a panel of 50 urinary, periurethral, and rectal E. coli isolates from patients with UTI. These studies outline a computational approach that may be broadly applicable for studying strain-specific adaptation and pathogenesis in other bacteria.