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Biomedical subjects

C Cherbut

Publications and source records attributed to C Cherbut.

At least 37 records · Page 2Linked to original sources

Short-chain fatty acids modify colonic motility through nerves and polypeptide YY release in the rat.

Short-chain fatty acids (SCFAs) are recognized as the major anions of the large intestinal content in humans, but their effect on colonic motility is controversial. This study explores the colonic motor effect of SCFAs and their mechanisms in the rat. Colonic motility (electromyography) and transit time (plastic markers) were measured in conscious rats while SCFAs were infused into the colon, either alone or after administration of neural antagonists or immunoneutralization of circulating polypeptide YY (PYY). SCFA-induced PYY release was measured by RIA and then simulated by infusing exogenous PYY. Intracolonic infusion of 0.4 mmol/h SCFAs had no effect, whereas 2 mmol/h SCFAs reduced colonic motility (36 +/- 3 vs. 57 +/- 4 spike bursts/h with saline, P < 0.05) by decreasing the ratio of nonpropulsive to propulsive activity. This resulted in an increased transit rate (P < 0.01). Neither alpha-adrenoceptor blockade nor nitric oxide synthase inhibition prevented SCFA-induced motility reduction. Intraluminal procaine infusion suppressed the SCFA effect, indicating that a local neural mechanism was involved. SCFA colonic infusion stimulated PYY release in blood. Immunoneutralization of circulating PYY abolished the effect of SCFAs on colonic motility, whereas exogenous PYY infusion partly reproduced this effect. SCFAs modify colonic motor patterns in the rat and increase transit rate; local nerve fibers and PYY are involved in this effect.

Animals↗

Butyrate stimulates cyclin D and p21 and inhibits cyclin-dependent kinase 2 expression in HT-29 colonic epithelial cells.

Sodium butyrate, a product of colonic fermentation of dietary fiber, has been shown to inhibit cell proliferation by blocking the cells in the G1 phase of the cell cycle. However, its mechanism of action is still unknown. We found that butyrate strongly stimulated cyclin D and p21/WAF1/CIP1 expression in HT-29 human colonic adenocarcinoma cells, in a dose dependent manner. These stimulations were associated with a decrease in cyclin-dependent kinase (cdk) 2 level, whereas cdk4 and cdk6 remained unchanged. Our results suggest that the inhibition of cell cycle progression by sodium butyrate may be explained by a modulation of cell cycle regulatory proteins such as cyclin D and p21.

Butyrates↗

Treatment of rat proximal and distal colonic cells with sodium orthovanadate enhances their adhesion and survival in primary culture.

We have studied the effect of sodium orthovanadate, an inhibitor of protein tyrosine phosphatases, on primary cultures of colonocytes and stromal cells. Everted proximal and distal colonic tissue of adult rats were disintegrated by a collagenase/dispase solution for 60 min at 37 degrees C to prepare viable gland fragments and isolated cells. Cell preparations were inoculated onto plastic substratum or cytodex-3 microcarriers in a defined maintenance medium or in 1% fetal calf serum media. Incorporation of sodium orthovanadate (> or = 50 microM) in these media constantly enhanced the survival (cell enumeration and trypan blue exclusion P < 0.05) and the adhesion (up to four-fold by crystal violet staining, P < 0.01) of colonocytes (characterized by cytokeratin-18, transforming growth factor-alpha or alkaline phosphatase expression) and stromal cells. Removal of sodium orthovanadate from culture media restored cellular death processes. Incorporation of 10 mM n-butyric acid did not promote cell adhesion and survival except for distal cells exposed to 2 mM sodium orthovanadate. Besides studies in the regulation of anoikis in primary culture, the model will help to assay the influences of dietary and growth factors on the biology of non-cancerous colonic cells.

Animals↗

Comparison of the effect of different short chain fatty acids on the growth and differentiation of human colonic carcinoma cell lines in vitro.

The aim of this study was to compare the effects of acetate, propionate, butyrate, iso-butyrate, valerate, iso-valerate and caproate on cell growth and on the activities of alkaline phosphatase (AP) and dipeptidyl aminopeptidase IV (DPP IV) by three human colonic adenocarcinoma cell lines. In addition to butyrate, propionate and valerate inhibited cell proliferation of the three cell lines. The other SCFAs did not influence cell proliferation. AP and DPP IV activities were strongly stimulated by butyrate on two of the three cell lines. On HT-29, AP was strongly stimulated, however DPPIV expression remained undetectable. Propionate and valerate exhibited a weaker stimulation, the other SCFAs being ineffective. The effect of SCFAs on cell proliferation and differentiation clearly depends on the number of carbons and on the configuration of the basic structure of the molecule.

Alkaline Phosphatase↗

Digestive and metabolic effects of potato and maize fibres in human subjects.

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.

Adult↗

Effects of short-chain fatty acids on gastrointestinal motility.

Besides their action on gut morphology and function, short-chain fatty acids (SCFAs), produced by bacterial fermentation of carbohydrates in the colon, influence gastrointestinal motility. As they are not present in the stomach and proximal small intestine, SCFAs do not directly affect motility of these segments. However, caecal infusion of SCFAs as well as colonic fermentation of lactulose induce a relaxation of the proximal stomach in humans, indicating that SCFAs can affect motility at a distance from their site of production. Moreover, this suggests that SCFAs may be involved in the so-called "ileocolonic brake', i.e. the inhibition of gastric emptying by nutrients reaching the ileo-colonic junction. In the terminal ileum, where their concentration may increase following a colo-ileal reflux, SCFAs stimulate contractions and shorten ileal emptying, which may protect ileal mucosa against the potentially harmful effects of the reflux of colonic contents. Although SCFAs are produced and concentrated in the colon, their action on motility of this organ is not clearly understood and may depend on concentration, molecular structure of the acids, responsiveness of the colonic segments and animal species. The mechanisms of action of SCFAs on gastrointestinal motility are not completely elucidated. They may involve systemic humoral and neural pathways as well as local reflexes and myogenic responses.

Animals↗

Specific adaptation of gastric emptying to diets with differing protein content in the rat: is endogenous cholecystokinin implicated?

BACKGROUND: Recent studies indicate that gastric emptying may be influenced by patterns of previous nutrient intake. Endogenous cholecystokinin (CCK), whose synthesis and release can be affected by dietary intake, has a major role in the regulation of gastric emptying. AIMS: To evaluate the influence of diets with differing protein content on gastric emptying of differing liquid test meals and plasma CCK levels in the rat and to check whether the inhibitory effect of exogenous CCK on gastric emptying is modified after long term intake of diets with differing protein content. METHODS: Rats were fed for three weeks with high protein, medium protein (regular), or low protein diet. On day 22 gastric emptying of a peptone meal was studied. In addition, basal and postprandial CCK levels after the different dietary regimens were measured by bioassay. The time course of dietary adaptation was studied and its specificity assessed through the use of different (peptone, glucose, and methylcellulose) test meals. The effect of exogenous CCK-8 on gastric emptying was studied at the end of the adaptation period (three weeks). RESULTS: Feeding the animals with a high protein diet for three weeks resulted in a significant (p < 0.05) acceleration (by 21.2 (8.2)%) of gastric emptying while feeding with a low protein diet was followed by a significant (p < 0.05) delay (by 24.0 (6.2)%) in the emptying rate. When the time course of the effect of dietary adaptation on gastric emptying was studied, it appeared that at least two weeks are required for dietary protein to be effective. The regulatory effect of dietary protein on gastric emptying proved to be dependent on meal composition. Only the emptying rate of a protein containing meal (40% peptone) was significantly modified by previous dietary intake. No significant (p > 0.05) changes were observed with glucose and methylcellulose meals whose emptying rates were similar in rats receiving a high protein or low protein diet. A peptone meal strongly and significantly (p < 0.05) increased plasma CCK levels in rats fed a medium protein (regular) diet. Results were similar in rats receiving a low protein diet (p < 0.05) but not in rats on a high protein diet (p > 0.05). As a consequence, postprandial plasma levels of CCK in rats fed with a medium or low protein diet were significantly (p < 0.05) higher than those in rats receiving a high protein diet. In rats on high and low protein diets, dose response curves to CCK-8 were virtually identical, suggesting that dietary protein intake has no influence on the effect of exogenous CCK. CONCLUSIONS: These results clearly show that gastric emptying of a protein containing meal can be modified by previous dietary protein intake. This effect, which is time dependent and meal specific, may be related to changes in endogenous CCK release which will affect emptying rate. While the exact mechanisms underlying this adaptive response need to be studied and clarified further, these results emphasise the importance of dietary history in the evaluation and interpretation of gastric emptying data.

Adaptation, Physiological↗

Butyrate enhances major histocompatibility complex class I, HLA-DR and ICAM-1 antigen expression on differentiated human intestinal epithelial cells.

Besides its metabolic role, butyrate, a by-product of colonic fermentation, can modulate colonocyte proliferation and the expression of various molecules. In inflammatory bowel diseases, epithelial cells express HLA class II molecules and may behave as antigen-presenting cells. This study was performed to characterize the effect of butyrate on major histocompatibility complex expression by human colonocytes in comparison with interferon-gamma. Five cell lines displaying different differentiation features were analysed for antigen expression by flow cytofluorimetry. All lines expressed class I antigens, whereas only SW 1116 cells express HLA-DR. On these cells, butyrate and interferon-gamma strongly enhanced HLA-DR and ICAM-1 expression, whereas a mild increase in class I antigens was noted. Moreover, an increase in class I antigens was observed on two other differentiated cell lines, and it was synergistic with interferon-gamma. Butyrate, by its modulation of HLA-DR, ICAM-1 and HLA class I expression, may act on antigen presentation and, thus, influence some inflammatory processes.

Acetates↗

Colonic fermentation and proximal gastric tone in humans.

BACKGROUND & AIMS: Some carbohydrates escape small intestinal absorption, and their presence in the ileum can affect proximal gut motility. Carbohydrates reaching the colon can inhibit gastric and pancreatic secretions. The hypothesis of this study was that colonic fermentation products of carbohydrates (short-chain fatty acids [SCFAs]) affect proximal gut motility and especially gastric tone. METHODS: Healthy volunteers were studied after oral administration of 20 g lactulose (n = 6) and intracolonic infusions of 20 g lactose (n = 7) and SCFAs (54 mmol/180 mL and 90 mmol/180 mL, respectively). Gastric tone (electronic barostat) and H2 concentrations in exhaled air were simultaneously monitored, and peripheral intestinal peptide levels were measured by specific radioimmunoassays. RESULTS: After oral lactulose administration (but not after saline), a significant decrease in gastric tone was observed, which rapidly followed the increase in H2 concentrations. Gastric tone also decreased after intracolonic infusions of both lactose and SCFAs; the most marked effect occurred after the highest SCFA dose. No significant changes in the level of plasma oxyntomodulin-like immunoreactivity and glucagon-like peptide 1 were found, whereas the level of peptide YY increased significantly over time, but not differently after saline and test solutions. CONCLUSIONS: Colonic fermentation of undigestible carbohydrates can inhibit gastric tone, and SCFAs may be responsible for this colonic brake. The role of intestinal peptides, if any, was not identified.

Adult↗

In vitro contractile effects of short chain fatty acids in the rat terminal ileum.

Short chain fatty acids (SCFAs), produced in the gut by bacterial fermentation of carbohydrates, change intestinal motility by mechanisms as yet unknown. This study examined the mechanism(s) of action of SCFAs on contractility using isolated rat terminal ileum segments and isolated ileal smooth muscle cells. Strip contractions were recorded under isometric conditions. Intracellular calcium concentration ([Ca2+]i) was measured in single cells loaded with indo-1 penta-acetoxymethyl ester (indo-1 AM). SCFAs (10(-9) to 10(-2) mol/l) induced concentration dependent contractions. The effect was not different among the individual SCFAs. Exogenous acids (namely tartaric and citric acids) caused similar responses as SCFAs, whereas sodium acetate had no effect. The contraction was not blocked by tetrodotoxin, atropine or hexamethonium, showing that it was not mediated through a cholinergic pathway. Moreover, removal of the mucosa or addition of procaine (a local anaesthetic) to the bath did not change the SCFA induced contraction, while verapamil (a calcium-channel antagonist) completely suppressed it. In addition, application of SCFAs to isolated ileal myocytes evoked peaks in [Ca2+]i inhibited by D 600 (a blocker of voltage dependent calcium channels). Taken together, these results suggest that the contractile response stimulated by SCFAs in the rat terminal ileum could result from an acid sensitive calcium dependent myogenic mechanism.

Acetates↗

Inhibition of acetylcholine induced intestinal motility by interleukin 1 beta in the rat.

BACKGROUND/AIMS: The fact that raised interleukin 1 beta (IL 1 beta) concentrations have been found in the colonic mucosa of rats with experimentally induced colitis and of patients with inflammatory bowel disease indicates that this cytokine may participate in the disturbed intestinal motility seen during inflammatory bowel disease. This study investigated whether IL 1 beta could change the contractility of (a) a longitudinal muscle-myenteric plexus preparation from rat jejunum, ileum, and colon and (b) isolated jejunal smooth muscle cells. METHODS: Isometric mechanical activity of intestinal segments was recorded using a force transducer. Moreover, smooth muscle cell length was measured by image analysis. RESULTS: Although IL 1 beta did not affect jejunal, ileal, and colonic basal contractility, it significantly reduced contractile response to acetylcholine (ACh). This significant inhibition was seen only after 90 or 150 minutes of incubation with IL 1 beta. Pretreatment with cycloheximide blocked IL 1 beta induced inhibition of ACh stimulated jejunal contraction, suggesting that a newly synthesised protein was involved in the effect. NW-nitro-L-arginine (a nitric oxide synthase inhibitor) did not prevent the inhibition induced by IL 1 beta. Blocking neural transmission with tetrodotoxin abolished the IL 1 beta effect on jejunal contractile activity, whereas IL 1 beta had no effect on isolated and dispersed smooth muscle cells. CONCLUSIONS: IL 1 beta inhibits ACh induced intestinal contraction and this inhibitory effect involves protein synthesis but is independent of nitric oxide synthesis. This effect does not involve a myogenic mechanism but is mediated through the myenteric plexus.

Animals↗

Involvement of small intestinal motility in blood glucose response to dietary fibre in man.

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.

Adult↗

Digestibility and bulking effect of ispaghula husks in healthy humans.

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.

Acetates↗

Influence of particle size and sources of non-starch polysaccharides on postprandial glycaemia, insulinaemia and triacylglycerolaemia in pigs and starch digestion in vitro.

Physico-chemical properties of dietary fibres might be involved in metabolic control, particularly of the postprandial blood glucose response. The aim of the present study was to look at the effects of the content of soluble fibres and of the particle size of solid fibres on in vitro and in vivo starch hydrolysis and on the subsequent glucose absorption as well as the triacylglycerolaemia. Two sources of dietary fibres, one, with soluble fibres (beet pulp), the other with mostly insoluble fibres (wheat bran), were added at the rate of 60 g/kg to a meal simulating human food. The fibre sources were ground in order to obtain two different particle sizes (250 and 500 microns). Beet pulp decreased significantly (P < 0.05) initial in vitro hydrolysis whereas wheat bran increased starch hydrolysis in the first 10 min. Wheat bran and beet pulp, whatever its particle size, lowered the post-prandial triacylglycerol response. No significant effect was found with dietary fibre-supplemented diets on postprandial glycaemic and insulinaemic values. High correlation was found between initial in vitro starch hydrolysis and mean areas under the insulinaemic curves. This in vitro model can be used to predict initial in vivo digestion of carbohydrates from complex foods.

Animals↗

Sugar composition of dietary fibre and short-chain fatty acid production during in vitro fermentation by human bacteria.

The aim of the present study was to assess the relationship between the disappearance of dietary fibre sugars and the production of individual short-chain fatty acids (SCFA). The bacterial degradation of five dietary fibres whose sugars were quantified was investigated in vitro using a human faecal inoculum. Involvement of the main fibre sugars in SCFA production was evaluated by a stepwise multiple linear regression. The results show first that the nature and chiefly the associations between the fibre sugars were key variables in the fermentability. Second, the nature and the amounts of SCFA produced were closely related to the in vitro fermentation of the main sugars available: uronic acids seemed to be principally involved in the production of acetic acid whereas the production of propionic acid could be promoted by the fermentation of glucose and, to a lesser extent, by that of xylose and arabinose. Xylose tended to have a greater impact than uronic acids and glucose on the production of butyric acid. Thus, it would be possible to predict which SCFA could be specifically produced during the fermentation of a fibre, as far as the chemical composition and structure of this fibre are known.

Acetates↗