RESUMEN
The perception of sweet is mediated by the sweet taste receptor T1R2-T1R3 expressed in taste cells of the lingual epithelium. This receptor is also expressed in intestinal enteroendocrine cells and is required for sensing luminal sugars and sweeteners to regulate expression of intestinal Na+-glucose cotransporter 1 (SGLT1). There are some notable differences amongst species in the ability to detect certain non-nutritive (artificial) sweeteners. Amino acid substitutions and pseudogenization of taste receptor genes are responsible for these disparities. Using heterologous expression, we demonstrate that the commonly used non-nutritive sweeteners sucralose, saccharin and acesulfame K activate pig T1R2-T1R3, but that aspartame and cyclamate do not. Furthermore, we show that in vitro sweetener activation of pig T1R2-T1R3 mirrors the sweetener stimulation of the gut-expressed receptor in vivo. Considering that sweeteners are included in animal feed worldwide, determination of taste receptor specificities in different species is essential for the development of scientifically-based dietary formulations.
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BACKGROUND: The composition of diets consumed following weight loss (WL) can have a significant impact on satiety and metabolic health. OBJECTIVE: This study was designed to test the effects of including a nondigestible carbohydrate to achieve weight maintenance (WM) following a period of WL. METHODS: Nineteen volunteers [11 females and 8 males, aged 20-62 y; BMI (kg/m2): 27-42] consumed a 3-d maintenance diet (15%:30%:55%), followed by a 21-d WL diet (WL; 30%:30%:40%), followed by 2 randomized 10-d WM diets (20%:30%:50% of energy from protein:fat:carbohydrate) containing either resistant starch type 3 (RS-WM; 22 or 26 g/d for females and males, respectively) or no RS (C-WM) in a within-subject crossover design without washout periods. The primary outcome, WM after WL, was analyzed by body weight. Secondary outcomes of fecal microbiota composition and microbial metabolite concentrations and gut hormones were analyzed in fecal samples and blood plasma, respectively. All outcomes were assessed at the end of each dietary period. RESULTS: Body weight was similar after the RS-WM and C-WM diets (90.7 and 90.8 kg, respectively), with no difference in subjectively rated appetite. During the WL diet period plasma ghrelin increased by 36% (P < 0.001), glucose-dependent insulinotropic polypeptide (GIP) decreased by 33% (P < 0.001), and insulin decreased by 46% (P < 0.001), but no significant differences were observed during the RS-WM and C-WM diet periods. Fasting blood glucose was lower after the RS-WM diet (5.59 ± 0.31 mmol/L) than after the C-WM diet [5.75 ± 0.49 mmol/L; P = 0.015; standard error of the difference between the means (SED): 0.09]. Dietary treatments influenced the fecal microbiota composition (R2 = 0.054, P = 0.031) but not diversity. CONCLUSIONS: The metabolic benefits, for overweight adults, from WL were maintained through a subsequent WM diet with higher total carbohydrate intake. Inclusion of resistant starch in the WM diet altered gut microbiota composition positively and resulted in lower fasting glucose compared with the control, with no apparent change in appetite. This trial was registered at clinicaltrials.gov as NCT01724411.
Asunto(s)
Fibras de la Dieta/farmacología , Microbioma Gastrointestinal , Sobrepeso/dietoterapia , Pérdida de Peso , Adulto , Bacterias/clasificación , Bacterias/genética , ADN Bacteriano/genética , Dieta Reductora , Fibras de la Dieta/administración & dosificación , Heces/microbiología , Femenino , Intolerancia a la Glucosa , Humanos , Lípidos/sangre , Masculino , Persona de Mediana Edad , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Adulto JovenRESUMEN
Mucosa-associated microbial populations of the gastrointestinal tract are in intimate contact with the outer mucus layer. This proximity offers these populations a higher potential, than lumenal microbiota, in exerting effects on the host. Functional characteristics of the microbiota and influences of host-physiology shape the composition and activity of the mucosa-associated bacterial community. We have shown previously that inclusion of an artificial sweetener, SUCRAM, included in the diet of weaning piglets modulates the composition of lumenal-residing gut microbiota and reduces weaning-related gastrointestinal disorders. In this study, using Illumina sequencing we characterised the mucosa-associated microbiota along the length of the intestine of piglets, and determined the effect of SUCRAM supplementation on mucosa-associated populations. There were clear distinctions in the composition of mucosa-associated microbiota, between small and large intestine, concordant with differences in regional oxygen distribution and nutrient provision by the host. There were significant differences in the composition of mucosa-associated compared with lumenal microbiota in pig caecum. Dietary supplementation with SUCRAM affected mucosa-associated bacterial community structure along the length of the intestinal tract. Most notably, there was a substantial reduction in predominant Campylobacter populations proposing that SUCRAM supplementation of swine diet has potential for reducing meat contamination and promoting food safety.
Asunto(s)
Microbioma Gastrointestinal , Mucosa Intestinal/microbiología , Porcinos/microbiología , Animales , Dieta , Suplementos Dietéticos , Tracto Gastrointestinal/microbiología , DesteteRESUMEN
Disruption in stable establishment of commensal gut microbiota by early weaning is an important factor in susceptibility of young animals to enteric disorders. The artificial sweetener SUCRAM [consisting of neohesperidin dihydrochalcone (NHDC) and saccharin] included in piglets' feed reduces incidence of enteric disease. Pyrosequencing of pig caecal 16S rRNA gene amplicons identified 25 major families encompassing seven bacterial classes with Bacteroidia, Clostridia and Bacilli dominating the microbiota. There were significant shifts in microbial composition in pigs maintained on a diet containing SUCRAM, establishing SUCRAM as a major influence driving bacterial community dynamics. The most notable change was a significant increase of Lactobacillaceae population abundance, almost entirely due to a single phylotype, designated Lactobacillus 4228. The sweetener-induced increase in Lactobacillaceae was observed in two different breeds of pigs signifying a general effect. We isolated Lactobacillus 4228, sequenced its genome and found it to be related to Lactobacillus amylovorus. In vitro analyses of Lactobacillus 4228 growth characteristics showed that presence of NHDC significantly reduces the lag phase of growth and enhances expression of specific sugar transporters, independently of NHDC metabolism. This study suggests that sensing of NHDC by a bacterial plasma membrane receptor underlies sweetener-induced growth of a health promoting gut bacterium.
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Bacterias/metabolismo , Microbioma Gastrointestinal , Intestinos/microbiología , Lactobacillus/crecimiento & desarrollo , Edulcorantes/metabolismo , Porcinos/microbiología , Alimentación Animal/análisis , Animales , Bacterias/genética , Bacterias/aislamiento & purificación , Ciego/metabolismo , Ciego/microbiología , Femenino , Mucosa Intestinal/metabolismo , Lactobacillus/metabolismo , Masculino , Sacarina/metabolismo , Porcinos/metabolismo , DesteteRESUMEN
The diet of the horse, pasture forage (grass), is fermented by the equine colonic microbiota to short-chain fatty acids, notably acetate, propionate and butyrate. Short-chain fatty acids provide a major source of energy for the horse and contribute to many vital physiological processes. We aimed to determine both the mechanism of butyrate uptake across the luminal membrane of equine colon and the nature of the protein involved. To this end, we isolated equine colonic luminal membrane vesicles. The abundance and activity of cysteine-sensitive alkaline phosphatase and villin, intestinal luminal membrane markers, were significantly enriched in membrane vesicles compared with the original homogenates. In contrast, the abundance of GLUT2 protein and the activity of Na(+)-K(+)-ATPase, known markers of the intestinal basolateral membrane, were hardly detectable. We demonstrated, by immunohistochemistry, that monocarboxylate transporter 1 (MCT1) protein is expressed on the luminal membrane of equine colonocytes. We showed that butyrate transport into luminal membrane vesicles is energized by a pH gradient (out < in) and is not Na(+) dependent. Moreover, butyrate uptake is time and concentration dependent, with a Michaelis-Menten constant of 5.6 ± 0.45 mm and maximal velocity of 614 ± 55 pmol s(-1) (mg protein)(-1). Butyrate transport is significantly inhibited by p-chloromercuribenzoate, phloretin and α-cyano-4-hydroxycinnamic acid, all potent inhibitors of MCT1. Moreover, acetate and propionate, as well as the monocarboxylates pyruvate and lactate, also inhibit butyrate uptake. Data presented here support the conclusion that transport of butyrate across the equine colonic luminal membrane is predominantly accomplished by MCT1.
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Butiratos/farmacocinética , Colon/metabolismo , Mucosa Intestinal/metabolismo , Intestino Grueso/metabolismo , Animales , Transporte Biológico , Membrana Celular/metabolismo , Colon/efectos de los fármacos , Transportador de Glucosa de Tipo 2/metabolismo , Caballos , Humanos , Mucosa Intestinal/efectos de los fármacos , Intestino Grueso/efectos de los fármacos , Transportadores de Ácidos Monocarboxílicos/metabolismo , Simportadores/metabolismoRESUMEN
The commensal bacteria Lactobacillus are widely used as probiotic organisms conferring a heath benefit on the host. They have been implicated in promoting gut health via the stimulation of host immunity and anti-inflammatory responses, as well as protecting the intestinalmucosa against pathogen invasion. Lactobacilli grow by fermenting sugars and starches and produce lactic acid as their primary metabolic product. For efficient utilisation of varied carbohydrates, lactobacilli have evolved diverse sugar transport and metabolic systems, which are specifically induced by their own substrates. Many bacteria are also capable of sensing and responding to changes in their environment. These sensory responses are often independent of transport or metabolism and are mediated through membrane-spanning receptor proteins. We employed DNA-based pyrosequencing technology to investigate the changes in the intestinal microbiota of piglets weaned to a diet supplemented with either a natural sugar, lactose or an artificial sweetener (SUCRAM®, consisting of saccharin and neohesperidin dihydrochalcone (NHDC); Pancosma SA). The addition of either lactose or saccharin/NHDC to the piglets' feed dramatically increased the caecal population abundance of Lactobacillus, with concomitant increases in intraluminal lactic acid concentrations. This is the first report of the prebiotic-like effects of saccharin/NHDC, an artificial sweetener, being able to influence the commensal gut microbiota. The identification of the underlying mechanism(s) will assist in designing nutritional strategies for enhancing gut immunity and maintaining gut health.
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Ciego/microbiología , Chalconas/farmacología , Hesperidina/análogos & derivados , Lactobacillus/crecimiento & desarrollo , Lactosa/farmacología , Prebióticos , Sacarina/farmacología , Edulcorantes/farmacología , Animales , Ciego/metabolismo , Suplementos Dietéticos , Fermentación , Hesperidina/farmacología , Ácido Láctico/metabolismo , Porcinos , DesteteRESUMEN
Luminal nutrient sensing by G-protein-coupled receptors (GPCR) expressed on the apical domain of enteroendocrine cells activates intracellular pathways leading to secretion of gut hormones that control vital physiological processes such as digestion, absorption, food intake and glucose homeostasis. The taste 1 receptor (T1R) family of GPCR consists of three members: T1R1; T1R2; T1R3. Expression of T1R1, T1R2 and T1R3 at mRNA and protein levels has been demonstrated in the intestinal tissue of various species. It has been shown that T1R2-T1R3, in association with G-protein gustducin, is expressed in intestinal K and L endocrine cells, where it acts as the intestinal glucose (sweet) sensor. A number of studies have demonstrated that activation of T1R2-T1R3 by natural sugars and artificial sweeteners leads to secretion of glucagon-like peptides 1&2 (GLP-1 and GLP-2) and glucose dependent insulinotropic peptide (GIP). GLP-1 and GIP enhance insulin secretion; GLP-2 increases intestinal growth and glucose absorption. T1R1-T1R3 combination co-expressed on the apical domain of cholecystokinin (CCK) expressing cells is a luminal sensor for a number of L-amino acids; with amino acid-activation of the receptor eliciting CCK secretion. This article focuses on the role of the gut-expressed T1R1, T1R2 and T1R3 in intestinal sweet and L-amino acid sensing. The impact of exploiting T1R2-T1R3 as a nutritional target for enhancing intestinal glucose absorption and gut structural maturity in young animals is also highlighted.
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Aminoácidos/metabolismo , Células Enteroendocrinas/metabolismo , Hormonas Gastrointestinales/metabolismo , Glucosa/metabolismo , Mucosa Intestinal/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Sensación , Animales , Células Quimiorreceptoras/metabolismo , Humanos , Intestinos/citología , Intestinos/crecimiento & desarrollo , Estado Nutricional , Transducción de Señal , Edulcorantes/metabolismo , Gusto , Transducina/metabolismoRESUMEN
Studies carried out in several species have demonstrated that detection of low-calorie sweeteners in the lumen of the intestine, by the sweet receptor, T1R2-T1R3, initiates a signaling pathway leading to enhanced expression and activity of intestinal Na+/glucose cotransporter 1, SGLT1. This results in an increased gut capacity to absorb glucose, sodium chloride and water, the basis for oral rehydration therapy. Horses express T1R2, T1R3 and downstream signaling elements in the intestinal tissue. As such, the potential of sweetener-stimulation of T1R2-T1R3 leading to upregulation of SGLT1 allows the provision of more glucose (energy) and hydration for horses. This is especially important when the need for glucose increases during strenuous exercise, pregnancy, and lactation. There are significant differences among species in the ability to detect sweeteners. Amino acid substitutions and pseudogenization of taste receptor genes underlie these variations. Nothing is known about the sweetener specificity of horse T1R2-T1R3. Using heterologous expression methodology, we demonstrate that sweeteners sucralose, stevia and neohesperidin dihydrochalcone (NHDC) activate horse T1R2-T1R3, but cyclamate does not. Determination of sweetener specificity of equine sweet receptor is crucial for developing suitable dietary additives to optimize glucose absorption, hydration and avoiding the intestinal disease brought about by microbial fermentation of unabsorbed carbohydrate reaching the large intestine.
RESUMEN
CCK is secreted by endocrine cells of the proximal intestine in response to dietary components, including amino acids. CCK plays a variety of roles in digestive processes, including inhibition of food intake, consistent with a role in satiety. In the lingual epithelium, the sensing of a broad spectrum of L-amino acids is accomplished by the heteromeric amino acid (umami) taste receptor (T1R1-T1R3). T1R1 and T1R3 subunits are also expressed in the intestine. A defining characteristic of umami sensing by T1R1-T1R3 is its potentiation by IMP or GMP. Furthermore, T1R1-T1R3 is not activated by Trp. We show here that, in response to L-amino acids (Phe, Leu, Glu, and Trp), but not D-amino acids, STC-1 enteroendocrine cells and mouse proximal small intestinal tissue explants secrete CCK and that IMP enhances Phe-, Leu-, and Glu-induced, but not Trp-induced, CCK secretion. Furthermore, small interfering RNA inhibition of T1R1 expression in STC-1 cells results in significant diminution of Phe-, Leu-, and Glu-stimulated, but not Trp-stimulated, CCK release. In STC-1 cells and mouse intestine, gurmarin inhibits Phe-, Leu-, and Glu-induced, but not Trp-stimulated, CCK secretion. In contrast, the Ca(2+)-sensing receptor antagonist NPS2143 inhibits Phe-stimulated CCK release partially and Trp-induced CCK secretion totally in mouse intestine. However, NPS2143 has no effect on Leu- or Glu-induced CCK secretion. Collectively, our data demonstrate that functional characteristics and cellular location of the gut-expressed T1R1-T1R3 support its role as a luminal sensor for Phe-, Leu-, and Glu-induced CCK secretion.
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Aminoácidos/farmacología , Colecistoquinina/metabolismo , Tracto Gastrointestinal/fisiología , Receptores Acoplados a Proteínas G/fisiología , Aminoácidos/antagonistas & inhibidores , Animales , Western Blotting , Membrana Celular/metabolismo , Células Cultivadas , Femenino , Tracto Gastrointestinal/efectos de los fármacos , Inmunohistoquímica , Fosfatos de Inositol/metabolismo , Isomerismo , Masculino , Ratones , Ratones Endogámicos C57BL , Microscopía Confocal , Naftalenos/farmacología , Péptidos/farmacología , Proteínas de Plantas/farmacología , Hidrolisados de Proteína/farmacología , Interferencia de ARN , Receptores Acoplados a Proteínas G/efectos de los fármacos , Estimulación QuímicaRESUMEN
The heteromeric sweet taste receptor T1R2-T1R3 is expressed on the luminal membrane of certain populations of enteroendocrine cells. Sensing of sugars and other sweet compounds by this receptor activates a pathway in enteroendocrine cells, resulting in secretion of a number of gut hormones, including glucagon-like peptide 2 (GLP-2). This subsequently leads to upregulation in the expression of intestinal Na(+)/glucose cotransporter, SGLT1, and increased intestinal glucose absorption. On the basis of the current information available on the horse genome sequence, it has been proposed that the gene for T1R2 (Tas1R2) is absent in the horse. We show here, however, that horses express both the mRNA and protein for T1R2. Equine T1R2 is most closely homologous to that in the pig and the cow. T1R2 protein, along with T1R3, α-gustducin, and GLP-2 proteins are coexpressed in equine intestinal endocrine cells. Intravenous administration of GLP-2, in rats and pigs, leads to an increase in the expression of SGLT1 in absorptive enterocytes and enhancement in blood glucose concentrations. GLP-2 receptor is expressed in enteric neurons, excluding the direct effect of GLP-2 on enterocytes. However, electric stimulation of enteric neurons generates a neural response leading to SGLT1 upregulation, suggesting that sugar in the intestine activates a reflex increase in the functional expression of SGLT1. Horses possess the ability to upregulate SGLT1 expression in response to increased dietary carbohydrates, and to enhance the capacity of the gut to absorb glucose. The gut sweet receptor provides an accessible target for manipulating the equine gut to absorb glucose (and water), allowing greater energy uptake and hydration for hard-working horses.
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Células Enteroendocrinas/metabolismo , Glucosa/metabolismo , Caballos/metabolismo , Intestino Delgado/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Secuencia de Aminoácidos , Animales , Transporte Biológico/efectos de los fármacos , Transporte Biológico/fisiología , Gatos , Carbohidratos de la Dieta/farmacocinética , Metabolismo Energético/efectos de los fármacos , Metabolismo Energético/fisiología , Células Enteroendocrinas/citología , Células Epiteliales/citología , Células Epiteliales/metabolismo , Femenino , Péptido 2 Similar al Glucagón/metabolismo , Intestino Delgado/citología , Masculino , Datos de Secuencia Molecular , Receptores Acoplados a Proteínas G/análisis , Transportador 1 de Sodio-Glucosa/metabolismo , Porcinos , Lengua/citología , Lengua/metabolismo , Transducina/metabolismoRESUMEN
We aimed to determine the effects of variations in dietary composition on equine gut microbiota and their fermentation products, and proposed that dietary modifications profoundly affect microbial ecosystems and their metabolites. Bacterial communities within the large intestine of three groups of horses were compared using oligonucleotide-RNA hybridisation methodology. Each group consisting of six horses was maintained on (1) a grass-only diet, (2) a concentrate diet (i.e. supplemented with hydrolysable carbohydrates) and (3) a concentrate diet but horses were affected by simple colonic obstruction and distension (SCOD), a prevalent form of dietary-induced intestinal disease. We show that in response to dietary change and intestinal disease, there is a progressive and significant increase in Lachnospiraceae, the Bacteroidetes assemblage and the lactic acid-producing, Bacillus-Lactobacillus-Streptococcus (BLS) group. In contrast, there is a corresponding decrease in the proportion of obligate fibrolytic, acid-intolerant bacteria, Fibrobacter and Ruminococcaceae. Assessment of monocarboxylic acids indicated that there are significantly higher concentrations of lactic acid in the colonic contents of horses maintained on a concentrate diet and those suffering from SCOD, correlating with the observed increase in the population abundance of the BLS group. However, the population size of the Veillonellaceae (lactate utilisers) remained constant in each study group. The inability of this group to respond to increased lactic acid may be a contributory factor to the build-up of lactic acid observed in horses fed a concentrate diet and those suffering from SCOD.
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Dieta , Caballos/metabolismo , Caballos/microbiología , Intestino Grueso/metabolismo , Intestino Grueso/microbiología , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Bacillus/aislamiento & purificación , Carga Bacteriana , Bacteroidetes/aislamiento & purificación , Fermentación , Fibrobacter/aislamiento & purificación , Enfermedades de los Caballos/metabolismo , Enfermedades de los Caballos/microbiología , Enfermedades Intestinales/metabolismo , Enfermedades Intestinales/microbiología , Enfermedades Intestinales/veterinaria , Lactobacillus/aislamiento & purificación , Metagenoma , ARN Bacteriano/genética , ARN Bacteriano/aislamiento & purificación , Streptococcus/aislamiento & purificaciónRESUMEN
Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Defective CFTR leads to accumulation of dehydrated viscous mucus within the small intestine, luminal acidification and altered intestinal motility, resulting in blockage. These changes promote gut microbial dysbiosis, adversely influencing the normal proliferation and differentiation of intestinal epithelial cells. Using Illumina 16S rRNA gene sequencing and immunohistochemistry, we assessed changes in mucosa-attached microbiome and epithelial cell profile in the small intestine of CF mice and a CF patient compared to wild-type mice and non-CF humans. We found increased abundance of pro-inflammatory Escherichia and depletion of beneficial secondary bile-acid producing bacteria in the ileal mucosa-attached microbiome of CFTR-null mice. The ileal mucosa in a CF patient was dominated by a non-aeruginosa Pseudomonas species and lacked numerous beneficial anti-inflammatory and short-chain fatty acid-producing bacteria. In the ileum of both CF mice and a CF patient, the number of absorptive enterocytes, Paneth and glucagon-like peptide 1 and 2 secreting L-type enteroendocrine cells were decreased, whereas stem and goblet cell numbers were increased. These changes in mucosa-attached microbiome and epithelial cell profile suggest that microbiota-host interactions may contribute to intestinal CF disease development with implications for therapy.
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Fibrosis Quística , Enfermedades Intestinales , Microbiota , Animales , Bacterias/genética , Recuento de Células , Fibrosis Quística/microbiología , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Células Caliciformes , Humanos , Enfermedades Intestinales/complicaciones , Mucosa Intestinal/microbiología , Intestino Delgado/microbiología , Ratones , ARN Ribosómico 16S/genéticaRESUMEN
In an intensive livestock production, a shorter suckling period allows more piglets to be born. However, this practice leads to a number of disorders including nutrient malabsorption, resulting in diarrhoea, malnutrition and dehydration. A number of strategies have been proposed to overcome weaning problems. Artificial sweeteners, routinely included in piglets' diet, were thought to enhance feed palatability. However, it is shown in rodent models that when included in the diet, they enhance the expression of Na+/glucose co-transporter (SGLT1) and the capacity of the gut to absorb glucose. Here, we show that supplementation of piglets' feed with a combination of artificial sweeteners saccharin and neohesperidin dihydrochalcone enhances the expression of SGLT1 and intestinal glucose transport function. Artificial sweeteners are known to act on the intestinal sweet taste receptor T1R2/T1R3 and its partner G-protein, gustducin, to activate pathways leading to SGLT1 up-regulation. Here, we demonstrate that T1R2, T1R3 and gustducin are expressed together in the enteroendocrine cells of piglet intestine. Furthermore, gut hormones secreted by the endocrine cells in response to dietary carbohydrates, glucagon-like peptides (GLP)-1, GLP-2 and glucose-dependent insulinotrophic peptide (GIP), are co-expressed with type 1 G-protein-coupled receptors (T1R) and gustducin, indicating that L- and K-enteroendocrine cells express these taste elements. In a fewer endocrine cells, T1R are also co-expressed with serotonin. Lactisole, an inhibitor of human T1R3, had no inhibitory effect on sweetener-induced SGLT1 up-regulation in piglet intestine. A better understanding of the mechanism(s) involved in sweetener up-regulation of SGLT1 will allow the identification of nutritional targets with implications for the prevention of weaning-related malabsorption.
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Suplementos Dietéticos , Intestino Delgado/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Transportador 1 de Sodio-Glucosa/metabolismo , Edulcorantes/farmacología , Porcinos/metabolismo , Transducina/metabolismo , Animales , Derivados del Benceno/farmacología , Transporte Biológico/efectos de los fármacos , Chalconas/farmacología , Carbohidratos de la Dieta/metabolismo , Células Enteroendocrinas/metabolismo , Femenino , Polipéptido Inhibidor Gástrico/metabolismo , Péptidos Similares al Glucagón/metabolismo , Hesperidina/análogos & derivados , Hesperidina/farmacología , Masculino , Sacarina/farmacología , Serotonina/metabolismo , Regulación hacia Arriba , DesteteRESUMEN
Na+/glucose co-transporter 1 (SGLT1) transports dietary sugars from the lumen of the intestine into enterocytes. Regulation of this protein is essential for the provision of glucose to the body and, thus, is important for maintenance of glucose homeostasis. We have assessed expression of SGLT1 at mRNA, protein and functional levels in the intestinal tissue of 28 d old piglets weaned onto isoenergetic diets with differing concentrations of digestible carbohydrate (CHO). We show that expression of SGLT1 remains constant when piglets are fed up to 40 % CHO-containing diets. However, there is a significant increase in SGLT1 expression when the CHO content of the diet is>50 %. Morphometric analyses indicate that the increased expression is not due to a trophic effect. It has been proposed that in rat intestine, in response to a high-CHO diet, GLUT2 (the classical basolateral membrane monosaccharide transporter) is translocated to the luminal membrane of enterocytes to absorb excess dietary glucose. We show, using immunohistochemistry and Western blotting with antibodies raised to amino acids in different epitopes of GLUT2, that under all dietary conditions, low to high CHO, GLUT2 is expressed on the basolateral membrane of pig enterocytes. Furthermore, functional studies indicate that there is no uptake of 2-deoxy-D-glucopyranoside, a specific substrate of Na+-independent glucose transporters into brush-border membrane vesicles isolated from the intestines of piglets either maintained on low- or high-CHO diets. Thus, SGLT1 is the major route for absorption of dietary sugars across the luminal membrane of swine enterocytes.
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Carbohidratos de la Dieta/administración & dosificación , Sacarosa en la Dieta/metabolismo , Enterocitos/metabolismo , Transportador de Glucosa de Tipo 2/metabolismo , Glucosa/metabolismo , Intestino Delgado/metabolismo , Transportador 1 de Sodio-Glucosa/metabolismo , Animales , Transporte Biológico , Membrana Celular/metabolismo , Epítopos , Femenino , Masculino , Microvellosidades/metabolismo , ARN/metabolismo , ARN Mensajero/metabolismo , Transportador 1 de Sodio-Glucosa/genética , DesteteRESUMEN
Absorption of glucose, via intestinal Na+/glucose cotransporter 1 (SGLT1), activates salt and water absorption and is an effective route for treating Escherichia coli (E. coli)-induced diarrhea. Activity and expression of SGLT1 is regulated by sensing of sugars and artificial/natural sweeteners by the intestinal sweet receptor T1R2-T1R3 expressed in enteroendocrine cells. Diarrhea, caused by the bacterial pathogen E. coli, is the most common post-weaning clinical feature in rabbits, leading to mortality. We demonstrate here that, in rabbits with experimentally E. coli-induced diarrhea, inclusion of a supplement containing stevia leaf extract (SL) in the feed decreases cumulative morbidity, improving clinical signs of disease (p < 0.01). We show that the rabbit intestine expresses T1R2-T1R3. Furthermore, intake of SL enhances activity and expression of SGLT1 and the intestinal capacity to absorb glucose (1.8-fold increase, p < 0.05). Thus, a natural plant extract sweetener can act as an effective feed additive for lessening the negative impact of enteric diseases in animals.
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Diarrea/veterinaria , Células Enteroendocrinas/metabolismo , Infecciones por Escherichia coli/veterinaria , Escherichia coli/fisiología , Mucosa Intestinal/efectos de los fármacos , Edulcorantes no Nutritivos/administración & dosificación , Extractos Vegetales/administración & dosificación , Conejos/microbiología , Transportador 1 de Sodio-Glucosa/metabolismo , Stevia/química , Animales , Diarrea/tratamiento farmacológico , Diarrea/microbiología , Diarrea/mortalidad , Células Enteroendocrinas/efectos de los fármacos , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/mortalidad , Femenino , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Masculino , Hojas de la Planta/química , Conejos/metabolismo , Transportador 1 de Sodio-Glucosa/genéticaRESUMEN
Experimental and epidemiological evidence suggests that consumption of hydrolyzable carbohydrate, hCHO (grain), by horses is an important risk factor for colic, a common cause of equine mortality. It is unknown whether the small intestinal capacity to digest hCHO and/or to absorb monosaccharides is limiting, or even if horses can adapt to increased carbohydrate load. We investigated changes in the brush-border membrane carbohydrate digestive enzymes and glucose absorptive capacity of horse small intestine in response to increased hCHO. Expression of the Na(+)/glucose co-transporter, SGLT1, was assessed by Western blotting, immunohistochemistry, Northern blotting, QPCR, and Na(+)-dependent D-glucose transport. Glucose transport rates, SGLT1 protein, and mRNA expression were all 2-fold higher in the jejunum and 3- to 5-fold higher in the ileum of horses maintained on a hCHO-enriched diet compared to pasture forage. Activity of the disaccharidases was unaltered by diet. In a well-controlled study, we determined SGLT1 expression in the duodenal and ileal biopsies of horses switched, gradually over a 2-month period, from low (<1.0 g/kg bwt/day) to high hCHO (6.0 g/kg bwt/day) diets of known composition. We show that SGLT1 expression is enhanced, with time, 2-fold in the duodenum and 3.3-fold in the ileum. The study has important implications for dietary management of the horse.
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Carbohidratos de la Dieta/farmacología , Transportador 1 de Sodio-Glucosa/metabolismo , Adaptación Fisiológica , Animales , Femenino , Transportador de Glucosa de Tipo 2/biosíntesis , Caballos , Íleon/efectos de los fármacos , Íleon/metabolismo , Inmunohistoquímica , Yeyuno/efectos de los fármacos , Yeyuno/metabolismo , Masculino , Microvellosidades/metabolismo , ARN Mensajero/metabolismo , Transportador 1 de Sodio-Glucosa/biosíntesis , Solubilidad , Sacarasa/metabolismo , alfa-Glucosidasas/metabolismoRESUMEN
The impact of dietary composition and prebiotics, in promoting the growth of beneficial groups of gut bacteria, is increasingly apparent. Using Illumina MiSeq sequencing of bacterial 16S rRNA genes, this study has aimed to characterize and compare the establishment of the gastrointestinal microbiota in dairy calves given two different commercial milk replacer (MR) diets. MR1 and MR2 contain different levels of macronutrients such as protein and fat. Moreover, differences in manufacturing methods infer that MR2 may contain a greater proportion of conjugated milk oligosaccharides (OS), while MR1 contains more free milk OS. A total of 10 dairy calves, five in each group, were assigned to one of the two MR diets. Freshly voided fecal samples were taken at 0, 7, 14, 28, and 49 days after first consumption of milk replacer. The relative abundance of two individual Bifidobacterium species, which are known to utilize milk OS, and Faecalibacterium prausnitzii were significantly higher at day 7 in the fecal microbiome of calves fed MR2 compared with MR1. These commensal bacteria are widely regarded as probiotic organisms that confer a health benefit on the host. Our findings suggest that the composition of bovine milk replacers can have significant effects on the establishment of the gut microbiota in pre-weaned (neonatal) dairy calves. Better understanding of milk composition-microbiota-host interactions in early life will inform targeted interventions to increase growth and reduce mortality in young animals.
RESUMEN
Fish production is increasingly important to global food security. A major factor in maintaining health, productivity and welfare of farmed fish is the establishment and promotion of a stable and beneficial intestinal microbiota. Understanding the effects of factors such as host and environment on gut microbial community structure is essential for developing strategies for stimulating the establishment of a health-promoting gut-microbiota. We compared intestinal microbiota of common carp and rainbow trout, two fish with different dietary habits, sourced from various farm locations. There were distinct differences in the gut microbiota of carp and trout intestine. The microbiota of carp was dominated by Fusobacteriia and Gammaproteobacteria, while the trout microbiota consisted predominantly of Mollicutes and Betaproteobacteria. The majority of bacterial sequences clustered into a relatively low number of operational taxonomic units (OTUs) revealing a comparatively simple microbiota, with Cetobacterium, Aeromonas and Mycoplasma being highly abundant. Within each species, fish from different facilities were found to have markedly similar predominant bacterial populations despite distinctly different rearing environments, demonstrating intra-species uniformity and significant influence of host selectivity. This study demonstrates that in fish the host species imparts substantial impact in shaping the community structure of the intestinal microbiota.
Asunto(s)
Conducta Alimentaria/fisiología , Microbioma Gastrointestinal , Animales , Bacterias/clasificación , Bacterias/genética , Biodiversidad , Carpas/microbiología , ADN Bacteriano/genética , Inglaterra , Explotaciones Pesqueras , Especificidad del Huésped , Oncorhynchus mykiss/microbiología , ARN Ribosómico 16S/genética , Alimentos Marinos/microbiología , Análisis de Secuencia de ADN/veterinariaRESUMEN
BACKGROUND & AIMS: Butyrate oxidation is impaired in intestinal mucosa of patients with inflammatory bowel diseases (IBD). Butyrate uptake by colonocytes involves the monocarboxylate transporter (MCT) 1. We aimed to investigate the role of MCT1 in butyrate oxidation deficiency during colonic inflammation. METHODS: Colonic tissues were collected from patients with IBD or healthy controls and from rats with dextran sulfate sodium (DSS)-induced colitis. The intestinal epithelial cell line HT-29 was treated with interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). MCT1 expression was analyzed by real-time reverse-transcription polymerase chain reaction, Western blot, and immunofluorescence. Butyrate uptake and oxidation in HT-29 cells was assessed using [(14)C]-butyrate. The mechanism of MCT1 gene regulation was analyzed by nuclear run-on and reporter gene assays. RESULTS: MCT1 messenger RNA (mRNA) and protein levels were markedly decreased in inflamed colonic mucosa of IBD patients and rats. In HT-29 cells, down-regulation of MCT1 mRNA and protein abundance by IFN-gamma and TNF-alpha correlated with a decrease in butyrate uptake and subsequent oxidation. IFN-gamma and TNF-alpha did not affect MCT1 mRNA stability but rather down-regulated gene transcription. We demonstrate that the cytokine response element is located in the proximal -111/+213 core region of the MCT1 promoter. CONCLUSIONS: The data suggest that butyrate oxidation deficiency in intestinal inflammation is a consequence of reduction in MCT1-mediated butyrate uptake. This reinforces the proposition that butyrate oxidation deficiency in IBD is not a primary defect.