RESUMO
AIM: To assess the role of lactate as a precursor for butyrate biosynthesis in human colonic microflora. METHODS AND RESULTS: Three human faecal microfloras were incubated in vitro with media supplemented with 30 mmol l(-1) unenriched or 13C-enriched lactate. Lactate metabolism and short-chain fatty acid (SCFA) production were quantified. Lactate conversion to butyrate was investigated by gas chromatography-mass spectrometry and the pathways involved were identified by 13C nuclear magnetic resonance spectroscopy. All human faecal microfloras rapidly and completely fermented lactate, yielding approx. 19 mmol l(-1) total SCFAs. However, the SCFA composition varied markedly between microfloras. Butyrate was the main end-product for two microfloras but not for the third (60 and 61%vs 27% of the net concentration of SCFA produced respectively). The latter was typified by its ability to produce propionate as a major product (37%), and valerate (3%). 13C-Labelling showed that butyrate was produced through the acetyl-CoA pathway and that the three microfloras possessed significant differences in their metabolic pathways for lactate consumption. CONCLUSIONS: In contrast to the ruminal microflora, the human intestinal microflora can utilize both d- and l-lactate as precursors for butyrate synthesis. Inter-individual variation is found. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that the butyrogenic capability of colonic prebiotics could be related to lactate availability. These findings will direct the development of selection strategies for the isolation of new butyrate-producing bacteria among the lactate-utilizing bacteria present in the human intestinal microfloras.
Assuntos
Ácido Butírico/metabolismo , Colo/microbiologia , Ácido Láctico/metabolismo , Adulto , Ácidos Graxos Voláteis/biossíntese , Fezes/microbiologia , Feminino , Fermentação/fisiologia , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Espectroscopia de Ressonância Magnética/métodos , Masculino , Pessoa de Meia-IdadeRESUMO
AIMS: In vitro studies have suggested that fructo-oligosaccharides (FOS) and resistant starch (two fermentable non-digestible carbohydrates) display different fermentation kinetics. This study investigated whether these substrates affect the metabolic activity and bacterial composition of the intestinal microflora differently depending on the caecocolonic segment involved. METHODS AND RESULTS: Eighteen rats were fed a low-fibre diet (Basal) or the same diet containing raw potato starch (RPS) (9%) or short-chain FOS (9%) for 14 days. Changes in wet-content weights, bacterial populations and metabolites were investigated in the caecum, proximal and distal colon and faeces. Both substrates exerted a prebiotic effect compared with the Basal diet. However, FOS increased lactic acid-producing bacteria (LAPB) throughout the caecocolon and in faeces, whereas the effect of RPS was limited to the caecum and proximal colon. As compared with RPS, FOS doubled the pool of caecal fermentation products, while the situation was just the opposite distally. This difference was mainly because of the anatomical distribution of lactate, which accumulated in the caecum with FOS and in the distal colon with RPS. Faeces reflected these impacts only partly, showing the prebiotic effect of FOS and the metabolite increase induced by RPS. CONCLUSIONS: This study demonstrates that FOS and RPS exert complementary caecocolonic effects. SIGNIFICANCE AND IMPACT OF THE STUDY: The RPS and FOS combined ingestion could be beneficial by providing health-promoting effects throughout the caecocolon.
Assuntos
Bactérias Anaeróbias/isolamento & purificação , Intestino Grosso/microbiologia , Oligossacarídeos/farmacologia , Amido/farmacologia , Animais , Bactérias Anaeróbias/efeitos dos fármacos , Bactérias Anaeróbias/metabolismo , Ceco/microbiologia , Colo/microbiologia , Contagem de Colônia Microbiana , Fermentação/efeitos dos fármacos , Ácido Láctico/biossíntese , Masculino , Ratos , Ratos Wistar , Solanum tuberosumRESUMO
Ulvans (from Ulva lactuca) constitute a dietary fiber structurally similar to the mammalian glycosaminoglycans but with unexplored biological or cytotoxic activities. From native low-viscosity preparations containing 33.5 molar % and 18.4 molar % of sulfate residues and uronic acid residues, respectively, we derived desulfated, reduced and desulfated-reduced polysaccharides with respectively 5.2, 2.9, and 4.5-4.9 molar % of sulfate residues and uronic acid residues. The effects of these preparations were examined on the adhesion, proliferation and differentiation of normal or tumoral colonic epithelial cells cultured in conventional (0.3-0.8 x 10(6) cells/ml) or rotating bioreactor (3-8 x 10(6) cells/ml) culture conditions. In conventional culture conditions, ulvan modified the adhesion phase and the proliferation of normal colonic cells and undifferentiated HT-29 cells according to their molecular weights and to the relative molar proportion of sulfate residues. From the native polysaccharides, we have screened sulfated ulvans (MW < 5,000) which inhibited the Caco-2 cell proliferation/differentiation program by inducing a low cell reactivity to Ulex europeaus-1 lectins in defined (p < 0.001) or serum-supplemented media (p < 0.01) but were inactive on normal colonocytes. In conclusion, this dietary fiber could be a source of oligosaccharides with a bioactivity, a cytotoxicity or a cytostaticity targeted to normal or cancerous epithelial cells.
Assuntos
Fibras na Dieta/farmacologia , Células Epiteliais/efeitos dos fármacos , Mucosa Intestinal/citologia , Extratos Vegetais/química , Polissacarídeos/farmacologia , Alga Marinha , Animais , Adesão Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Neoplasias do Colo , Ensaio de Unidades Formadoras de Colônias , Células Epiteliais/citologia , Humanos , Mucosa Intestinal/efeitos dos fármacos , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Ratos , Ácidos Sulfúricos , Células Tumorais CultivadasRESUMO
Some data suggest that the colonic microflora may adapt to produce more butyrate if given time and the proper substrate. To test this hypothesis, we investigated the effect of prolonged feeding of resistant potato starch on butyrate production. Rats were fed on either a low-fibre diet (basal) or the same diet supplemented with 90 g resistant potato starch/kg (PoS) for 0.5, 2 and 6 months. Short-chain fatty acid (SCFA) concentrations were determined in caecal and colonic contents at the end of each ingestion period. Total SCFA concentration increased over time throughout the caecocolonic tract with PoS, but was not modified with the basal diet. While propionate concentration was unchanged, butyrate concentration was highly increased by PoS at each time period in both the caecum and colon. Moreover, the butyrogenic effect of PoS increased over time, and the amount of butyrate was increased 6-fold in the caecum and proximal colon and 3-fold in the distal colon after 6 months compared with 0.5 months. Accordingly, the ratio butyrate:- total SCFA increased over time throughout the caecocolonic tract (12.6 (SE 2.8) v. 28 (SE 1.8)% in the caecum, 10.5 (SE 1.4) v. 26.8 (SE 0.9)% in the proximal colon, and 7.3 (SE 2.4) v. 23.9 (SE 2.7)% in the distal colon at 0.5 v. 6 months respectively), while the proportion of acetate decreased. Neither the proportion nor the concentration of butyrate was modified over time with the basal diet. Butyrate production was thus promoted by long-term ingestion of PoS, from the caecum towards the distal colon, which suggests that a slow adaptive process occurs within the digestive tract in response to a chronic load of indigestible carbohydrates.
Assuntos
Butiratos/metabolismo , Fibras na Dieta/administração & dosagem , Intestino Grosso/metabolismo , Solanum tuberosum , Amido/administração & dosagem , Animais , Ceco/metabolismo , Ceco/microbiologia , Colo/metabolismo , Colo/microbiologia , Contagem de Colônia Microbiana , Ácidos Graxos Voláteis/análise , Fezes/química , Fermentação , Intestino Grosso/microbiologia , Ácido Láctico/análise , Masculino , Ratos , Ratos Wistar , Fatores de TempoRESUMO
The physiological effects of dietary fibres in humans are due to their physico-chemical properties. However, it is difficult to predict these effects simply by measuring certain characteristics in vitro. Studies in human subjects are still required to assess the effectiveness of new substrates. The aim of the present study in healthy human subjects was to evaluate the effects of two novel fibres, potato (PF) and maize (MF), on fasting and postprandial blood concentrations of carbohydrate and lipid metabolites as well as on stool output and transit time. The chemical composition, water-binding capacity (WBC) and fermentative properties of the fibres were also characterized in order to determine their possible involvement in digestive and metabolic effects. Stools, as well as breath and blood samples, were collected after consumption for 1 month of either a basal diet (control) or a basal diet supplemented with fibre (15 g/d). MF resisted fermentation better than PF and had lower digestibility. However, both fibres increased faecal output of dry matter, neutral sugars and water. There was an inverse relationship between stool weight and orofaecal transit time, although only MF significantly reduced transit time. Orocaecal transit was lengthened by PF, probably because of its high WBC. PF ingestion also decreased postprandial plasma levels of total and esterified cholesterol but had no effect on fasting concentrations. In contrast, MF lowered fasting cholesterolaemia and increased free:esterified cholesterol. These particular physiological and fermentative properties suggest that PF and MF would be suitable ingredients in a healthy diet.
Assuntos
Carboidratos/sangue , Fibras na Dieta/administração & dosagem , Digestão/fisiologia , Lipídeos/sangue , Solanum tuberosum , Zea mays , Adulto , Colesterol/sangue , Jejum/fisiologia , Ácidos Graxos Voláteis/metabolismo , Fezes/química , Feminino , Trânsito Gastrointestinal , Humanos , Masculino , Pessoa de Meia-Idade , Período Pós-Prandial , Água/metabolismoRESUMO
The digestibility of ispaghula, a mucilage from Plantago ovata composed mainly of arabinoxylans, and its faecal bulking effect were studied in seven healthy volunteers who ingested a low fibre controlled diet plus either placebo or 18 g/day of ispaghula for two 15 day periods. Whole gut transit time and gas excretion in breath and flatus were not different during the periods of ispaghula and placebo ingestion. Faecal wet and dry weights rose significantly, however, during ispaghula ingestion. Faecal short chain fatty acid concentrations and the molar proportions of propionic and acetic acids also increased. Most of the ispaghula had reached the caecum four hours after ingestion in an intact highly polymerised form. During ispaghula ingestion, the increase in the faecal output of neutral sugars was accounted for by the faecal excretion of arabinose and xylose in an intact highly polymerised form; the apparent digestibilities of these sugars were 24 (11) and 53% (6) respectively (mean (SEM)). In conclusion, ispaghula is more resistant to fermentation than previously reported in humans, and its bulking effect largely results from intact material.
Assuntos
Fibras na Dieta , Trânsito Gastrointestinal , Psyllium , Acetatos/análise , Adulto , Arabinose/análise , Testes Respiratórios , Estudos Cross-Over , Digestão , Ácidos Graxos Voláteis/análise , Fezes/química , Feminino , Gases/análise , Humanos , Masculino , Propionatos/análise , Xilose/análiseRESUMO
Three dietary fibres with different physicochemical properties were studied in healthy humans for their effects on small intestinal motility and postprandial hyperglycaemia. Duodeno-jejunal motor activity was evaluated electromyographically for 180 min in six subjects who had ingested a test meal composed of glucose alone or glucose with 15 g of wheat bran (WB), sugar beet (SB) or ispaghula (I) fibres. Glucose and insulin concentrations were determined during the same period. Each subject received each of the four test meals randomly during a 4 d period. Addition of SB or I to the glucose meal altered duodeno-jejunal motility. Both of these fibres inhibited stationary contractile activity and increased the propagation length and velocity of propagated activity, whereas addition of WB had no effect. These results could reflect the high water-holding capacity of SB and I. Blood glycaemic response to the glucose meal was reduced by SB and I but remained unchanged with WB. Postprandial blood glucose levels were significantly correlated with the total motility index (r 0.82) and stationary activity (r 0.79). Taken together, these observations suggest that the contractile activity induced by dietary fibre in the small intestine probably plays a major role in delayed glucose absorption.
Assuntos
Glicemia/metabolismo , Fibras na Dieta , Motilidade Gastrointestinal/fisiologia , Intestino Delgado/fisiologia , Adulto , Glicemia/análise , Eletromiografia , Glucose/administração & dosagem , Humanos , Insulina/sangue , Absorção Intestinal/fisiologia , Masculino , Psyllium , Distribuição Aleatória , Triticum , VerdurasRESUMO
The absorption of xylose at different levels of the intestine was compared in five dogs receiving diets containing either wheat bran, polyethylene particles (PE), or horse-bean hulls. The absorption was determined by serial collection of the interstitial fluid (ISF) in different parts of the small intestine and colon, and blood concentration after the administration of D-xylose as a solution (0.5 g/kg body weight) into the duodenal bulb. Xylose was mostly absorbed from the duodenum, and its concentration in the duodenal ISF and in plasma was reduced on a diet containing fiber, irrespective of the nature of fiber. In contrast, a negative linear relation between the mean retention time of digesta in the small intestine and the amount of xylose absorbed by duodenum was evidenced (r = -0.843). The results indicate that changes in transit linked to the presence of fiber in a diet are a major operative factor in the rate of carbohydrate absorption. They suggest that the absorption can be affected by a relatively minor change in the intestinal transit of digesta.