Plasma ghrelin response following a period of acute overfeeding in normal weight men.

BACKGROUND: Ghrelin, a 28 amino-acid peptide secreted primarily from the stomach has been identified as the endogenous ligand for the growth hormone secretagogue receptor. Ghrelin is suppressed in the postprandial state and has been linked to both type II diabetes and obesity. AIMS: To investigate the effects of a period of overfeeding with high-fat dietary supplements on plasma ghrelin levels in nonobese men. METHODS: Six healthy males (21-34 y; BMI 21-24 kg/m(2)) underwent the dietary intervention after completing diet and exercise diaries for 7 days. For 3 further weeks subjects followed their own diet diary supplemented with 125 ml single cream and 50 g roasted peanuts (88 g fat, 15 g Protein, 8 g carbohydrate) every day. Oral fat tolerance tests (OFTT) were undertaken at baseline, 7, 14 and 21 days of fat supplementation. The diet was increased in energy by 3.9 MJ/day and from a mean of 29-45% energy intake from fat with a small weight gain noted each week (P=0.009). RESULTS: Ghrelin concentrations were significantly reduced during the baseline OFTT. The postprandial ghrelin response (AUC) was significantly reduced following 2 weeks of dietary supplementation (P=0.005) increasing the suppression of plasma ghrelin by 18% despite only a 3% increase in body weight. Plasma triacylglycerol (P=0.009) and leptin (P=0.035) concentrations were also elevated and postprandial pancreatic polypeptide levels decreased (P=0.038) following dietary-supplementation. CONCLUSIONS: These results suggest that the metabolic profile associated with obesity, including a reduction in plasma ghrelin levels, may be related to recent dietary energy intake and precedes the development of significant adiposity.

An investigation into the reversibility of the morphological and cytokinetic changes seen in the small intestine of riboflavin deficient rats.

BACKGROUND: Impaired iron handling in riboflavin deficiency is thought to be partially a result of significant morphological and cytokinetic changes within the small intestine. AIMS: The aim of the study was to find out if the responses of the rat small intestine to riboflavin deficiency induced at weaning could be reversed upon repletion. SUBJECTS: 48 female weanling Wistar rats were used for the purpose of the study. METHODS: Rats were fed a riboflavin deficient diet or a complete control diet for a period of five weeks followed by a repletion period of up to three weeks. Rats were killed on day 0, 2, 7, or 21 of repletion. The duodenum was removed and fixed for subsequent analysis. RESULTS: Five weeks of riboflavin deficiency significantly changed the morphology and cytokinetics of the duodenum; the changes were not reversed within the 21 day repletion period despite biochemical evidence for a correction of the deficiency. CONCLUSIONS: The results show that the small intestine cannot readily recover from a period of riboflavin deficiency induced at weaning, supporting the notion that the weaning period is a critical time for gastrointestinal development and highlighting the importance of adequate nutrition during infancy.

Cytokinetic and structural responses of the rat small intestine to riboflavin depletion.

The impaired absorption and metabolism of Fe seen in riboflavin deficiency is attributed, at least in part, to a hyperproliferative response in the small intestine, associated with an altered morphology. Studies were conducted in female weanling Wistar rats to explore further the effect of riboflavin deficiency on the cytokinetics and structure of the small intestine. Feeding a riboflavin-deficient diet for 8 weeks from weaning resulted in a significantly lower villus number, a significant increase in villus length and an increased rate of transit of enterocytes along the villi, compared with weight-matched controls. A second experiment focused on the 3 weeks after weaning and showed that riboflavin deficiency inhibits the increase in villus number observed in control animals over this period. We suggest that riboflavin deficiency induced at weaning impairs the normal increase in villus number and that prolonged deficiency leads to an adaptive increase in length of villi and depth of crypts.

Adaptation to high-fat diets: effects on eating behaviour and plasma cholecystokinin.

Twelve male subjects took part in a study to investigate the effects of overfeeding a high-fat diet (19.17 MJ/d; 58% energy from fat) for 2 weeks on plasma cholecystokinin (CCK) levels, food intake, and subjective feelings of hunger and fullness. Before and after the diet, subjects completed a 2-week weighed dietary inventory, formal measurements of food intake from a pre-selected appetizing evening meal were carried out, and blood samples were taken after a standard breakfast for measurement of CCK. Hunger and fullness were rated on visual analogue scales before and after each of these meals and at evening meals during the diet period. Following the high-fat diet there was a small non-significant increase in food intake from the pre-selected meal (6919 (SE 615) kJ v. 6405 (SE 540) kJ; P = 0.1) and a significant increase in the average daily food consumption measured from the diaries (10.25 (SE 0.49) MJ/d v. 9.59 (SE 0.62) MJ/d; P = 0.05). Corresponding trends of increasing feelings of hunger and declining fullness also occurred over the study period. Plasma CCK responses to the standard breakfast were raised following the diet (1285 (SE 153) v. 897 (SE 78) pM min; 3 h integrated CCK production post v. pre diet; P < 0.01) with the major differences observed at 90 and 120 min following the meal. These results suggest that the increase in food intake may be related to a down-regulation in putative CCK receptors responsible for food intake. Elevated CCK levels might suggest a corresponding down-regulation in CCK receptors responsible for feedback inhibition of CCK release.

Morphological changes in the rat small intestine in response to riboflavin depletion.

Female Wistar rats were weaned onto a diet deficient in riboflavin and compared with weight-matched and ad lib.-fed controls. The effects of riboflavin deficiency on villus morphometry and enterocyte number on the villi in the upper small intestine were studied. Riboflavin depletion was associated with increased villus length and a proportional increase in the number of cell positions along the villi. The total DNA, RNA and protein contents in the intestinal mucosa were not significantly different between any of the groups. Villus hypertrophy in the absence of increased cell number in the small intestine suggests that villus number may be reduced in riboflavin deficiency. Riboflavin deficiency did not influence the number of mucus-producing goblet cells or the amount of mucosal glycoprotein in the small intestine. Impaired production of mucus appeared not to be involved in the structural and functional changes seen in riboflavin deficiency.

Gastrointestinal adaptation to enhanced small intestinal lipid exposure.

Studies were performed on 20 male adult rats to investigate the effects of chronic intermittent infusion of lipid and physiological emulsifier into the distal small intestine on stomach to caecum transit time (SCTT) of the head of a test meal. SCTT was measured using environmental hydrogen analysis. Ileal lipid infusion normally delays gastric emptying and small intestinal transit (p < 0.001), but chronic intermittent infusion of lipid, given three times a week gradually reduced the delay in transit time until by four weeks it was no longer than control values. The lipid induced delay did not return during the four weeks after the chronic infusion had finished. Intermittent infusion of physiological emulsifier into the distal small intestine for four weeks did not change the control SCTT or the acute response to an ileal lipid infusion. SCTT of the head of the meal did not change in the four weeks after the physiological emulsifier infusion had stopped. In conclusion these results show that infusing rats intermittently with lipid for four weeks results in desensitisation of the mechanisms by which distal small intestinal lipid regulate SCTT of the head of a meal. This adaptation is not reversed within four weeks of withdrawal of the lipid infusion. These results emphasise the importance of assessing recent dietary history when assessing gastric emptying and small bowel transit times.

The effect of the cholecystokinin antagonist devazepide (L364718) on the ileal brake mechanism in the rat.

Studies were carried out on 28 male adult rats to investigate whether the selective cholecystokinin-receptor antagonist devazepide influences gastrointestinal transit under control conditions and when it is delayed by ileal infusion of lipid. Stomach-to-caecum transit time of the head of the test meal was measured using environmental hydrogen analysis and the distribution of the meal was assessed using the radiolabelled meal technique. Oral administration of devazepide (4 mg kg-1) had no significant effect on transit time of the head of the baked bean test meal under control conditions, but significantly reversed the delay in transit time induced by ileal infusion of lipid (P < 0.01). Studying the distribution of the meal showed that Intralipid delayed transit time by delaying both gastric emptying (P < 0.01) and small bowel transit (P < 0.05). Devazepide did not alter the control distribution of the meal during ileal saline infusion, but during ileal infusion of lipid, devazepide further delayed gastric emptying (P < 0.01); the geometric centre of the meal was situated more proximally in the gastrointestinal tract (P < 0.05), but there was more of the meal in the colon (P < 0.01). The latter is compatible with the early rise in environmental hydrogen during devazepide administration and ileal lipid infusion and suggests that peripheral cholecystokinin receptors may modulate or mediate the delay in small bowel transit induced by ileal lipid. However, the data also suggest that mechanisms other than those involving cholecystokinin play a dominant role in the regulation of postprandial and lipid-delayed gastric emptying of a meal.

The effect of an opiate receptor antagonist on the ileal brake mechanism in the rat.

Studies investigated the effect of the opiate antagonist naloxone (10 mg/kg) on stomach to caecum transit (SCTT) during ileal infusion of saline or Intralipid. SCTT of the head of the meal was measured by hydrogen analysis and meal distribution by the radiolabelled meal technique. Intralipid delayed SCTT by delaying both gastric emptying (p < 0.01) and small bowel transit. Naloxone did not affect SCTT during ileal saline infusion, but produced a distal shift (p < 0.05) in the geometric centre of the meal and increased radioactivity in the caecum (p < 0.001) 100 min after gavage. Naloxone abolished the delayed SCTT of the meal induced by ileal lipid infusion, with an associated increase in radioactivity in the caecum at 200 min (p < 0.01), although the geometric centre was shifted proximally within the intestine (p < 0.01). The results suggest that the ileal brake is mediated in part by endogenous opiate pathways.

In vitro effects of sennoside on contractile activity and fluid flow in the perfused large intestine of the rat.

The effect of sodium rhein on contractile activity and fluid flow in the rat complete large intestine was studied in vitro. Contractile activity was recorded using serosal strain gauges and volume transducers recorded distal fluid flow from the segment. Luminal sodium rhein (1 mM) produced a protracted increase in caecal activity yet increased colonic contractility transiently. Fluid flow from the preparation was increased and the number of propagated complexes was elevated after the initial 10 min of exposure. The effect did not appear to be related directly to dose. Sodium rhein (0.1 mM) did not significantly stimulate contractility and a higher dose (5 mM) only produced a transient effect on propagated contractions. However, this dose had the effect of significantly reducing activity when the rhein was replaced by normal buffer. The data suggest that the action of sodium rhein is subtle; after an initial excitation, the glycoside shifts the pattern of motor activity in favour of propulsion at the expense of segmentation. The large intestine is more able, therefore, to expel luminal contents in a caudal direction following the addition of this anthraquinone laxative.

Is cholecystokinin a satiety hormone? Correlations of plasma cholecystokinin with hunger, satiety and gastric emptying in normal volunteers.

This study was designed to test the putative role of plasma cholecystokinin (CCK) in eating behaviour by examining the relationships between bioassayed plasma CCK concentrations and rated sensations of satiety and hunger following the ingestion of an appetizing nutrient-dense meal and between plasma profiles of CCK and scintigraphically determined gastric emptying in nine healthy male volunteers. Mean plasma CCK levels from the whole group showed a significant negative correlation with mean hunger ratings (r = -0.64) and a positive correlation with fullness (r = +0.68). However, there was considerable interindividual variation and negative correlations with hunger were only seen in three out of nine subjects and positive correlations with fullness in four out of nine subjects. In contrast, there was a strong relationship between the plasma CCK produced over the first 2 h after the meal and the half time for gastric emptying of the meal (r = -0.81). These data do not support a direct role for circulating levels of CCK in the control of hunger and satiety following a meal. Instead, the strong correlation with gastric emptying is compatible with release of CCK in response to nutrient delivery into the small intestine.

Granisetron and ondansetron: effects on the ileal brake mechanism in the rat.

Studies were carried out on 20 male adult rats to investigate how the action of the selective 5-HT3-receptor antagonists, granisetron and ondansetron, influence gastrointestinal transit under control conditions and when stomach-to-caecum transit was delayed by ileal infusion of lipid. Stomach-to-caecum transit time (SCTT) was measured using environmental hydrogen analysis. Subcutaneous administration of granisetron (BRL 43694, 40, 80 or 150 micrograms kg-1) significantly delayed the passage of the head of the baked bean meal through the stomach and the small intestine under control conditions (P < 0.05). Similarly, subcutaneous administration of ondansetron (GR 38032F, 80 or 150 micrograms kg-1) delayed control SCTT of the head of the meal but this did not reach statistical significance. In contrast, granisetron significantly reversed the delay in SCTT induced by ileal infusion of lipid at 40 (P < 0.001), 80 (P < 0.01) and 150 micrograms kg-1 (P < 0.05). Ondansetron also reversed the lipid-induced delay at 40 (P < 0.01), 80 (P < 0.001) and 150 micrograms kg-1 (P < 0.001). These apparently conflicting results may be rationalized by postulating the presence of 5-HT3 receptors on afferent nerves which, when inhibited by the specific antagonists, initiate reflexes that both accelerate and delay transit.

Preliminary studies on the gastrointestinal responses to fatty meals in obese people.

Studies were carried out on eight obese (BMI 30-34.6 kg/m2) and seven age and sex-matched normal weight volunteers (BMI 20-25 kg/m2) to investigate the gastric emptying, mouth to caecum transit time (MCTT), plasma cholecystokinin (CCK) and sensory responses to high (30 g margarine; 1327 kJ (317 kcal)) and low (301 kJ (72 kcal)) fat soups. Gastric emptying was measured by gamma scintigraphy, MCTT was measured by the breath hydrogen technique, plasma CCK was measured using a bioassay technique and subjective sensations were recorded on visual analogue scales. The high fat meal emptied more slowly than the low fat meal in both the normal subjects (t1/2 = 86.3 +/- 9.2 vs. 36.7 +/- 2.8 min) but there were no differences in the emptying of either meal between the two groups of subjects. Increasing the fat content of the meal did not affect the mouth to caecum transit time (MCTT) in either group, nor were there differences between the groups in MCTT (180 +/- 23 vs. 188 +/- 35 min, normal vs. obese MCTT after low fat soup; 228 +/- 17 vs. 227 +/- 29 min, normal vs. obese MCTT after high fat soup). Despite similar rates of gastric emptying, the obese group showed a higher CCK production following the high fat meal than the normal weight group (540.4 +/- 65.9 vs. 336.9 +/- 51.4 pmol.min, 2 h integrated CCK production, obese vs. normal; P < 0.05). The obese group also reported feeling less hungry throughout the study than the controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of short-chain fatty acids on contractile activity and fluid flow in rat colon in vitro.

The effect of short-chain fatty acids (SCFAs) on the contractile activity and fluid output of the large bowel of the rat was studied using an isolated segment of cecum and colon, mounted in vitro. The rate of contractile activity per minute in the proximal, mid, and distal regions of the colon was depressed by luminal infusion of associated SCFAs either as a mixture (acetic, propionic, and butyric) or individually (100 mM/pH = 4.1, in each case). Dose responses were observed for the individual fatty acids, with the 100 mM solutions eliciting a more prominent reduction in colonic motor activity than that induced by 10 mM. Neither the Na salt of the fatty acids nor an acidified Krebs solution (pH = 4.1) inhibited contractile activity or fluid output. No reduction in the rate of contractile activity was observed in the cecum with any test solutions, except 100 mM butyric acid. The data suggest that SCFAs inhibit smooth muscle contractility and resultant fluid transit.

The effect of adrenoceptor antagonists on the ileal brake mechanism in the rat.

1. Studies were carried out in the rat to investigate the effect of adrenoceptor antagonists on stomach to caecum transit time under control conditions and during ileal infusion of Intralipid. Stomach to caecum transit time (SCTT) of the head of the meal was measured by use of environmental hydrogen analysis and the distribution of the meal was assessed by a scintigraphic technique. 2. Four adrenoceptor antagonists were used in these studies, the alpha 1 antagonist prazosin, the alpha 2 antagonist, idazoxan, the beta 1 antagonist atenolol and the beta 2 antagonist ICI 118551. 3. None of the antagonists affected SCTT of the head of the meal during ileal infusion of saline. However, the alpha 1 and beta 1 antagonists significantly reversed (P less than 0.05) the delay in SCTT induced by ileal infusion of Intralipid whereas the alpha 2 antagonist, idazoxan, potentiated this delay (P less than 0.05). 4. Study of the distribution of the radiolabelled meal showed that the Intralipid delayed SCTT by slowing both gastric emptying (P less than 0.05) and small bowel transit (P less than 0.05). 5. Prazosin delayed gastric emptying under control conditions (P less than 0.001) but did not alter significantly the effect of ileal lipid on the distribution of the meal, 100 min or 200 min after gavage.6. The meal distribution was more compatible with the hydrogen analysis after administration of the ,beta-adrenoceptor antagonists. The reversal of the lipid-induced delay in SCTT caused by atenolol was associated with more radioactivity in the large intestine 200min after the gavage. ICI 118551 had no significant effects on either the distribution of the meal or the SCTT of the head of the meal.7. In conclusion, the data confirm that the sympathetic nervous system normally modulates or mediates the mechanisms that influence gastrointestinal transit in the rat and suggest that these mechanisms may be involved in the ileal brake effect. Nevertheless the data also suggest that simple measurement of the transit of the head of the meal by use of environmental hydrogen analysis may sometimes give a misleading impression of the action of drugs on gastrointestinal transit of the bulk of a test meal.

The effect of a 5-HT3-antagonist on the ileal brake mechanism in the rat.

Studies have been carried out on 7 male adult rats to investigate how the action of the selective 5-HT3 receptor antagonist, granisetron, influences gastrointestinal transit under control conditions and when it is delayed by ileal infusion of lipid. Stomach to caecum transit time (SCTT) was measured using environmental hydrogen analysis. Subcutaneous administration of granisetron (BRL 43694, 40 micrograms kg-1) significantly delayed the passage of the head of the baked bean meal through the stomach and the small intestine under control conditions (P less than 0.05). The same compound, however, significantly reversed the delay in SCTT induced by ileal infusion of lipid (P less than 0.001). These apparently paradoxical results may be rationalized by postulating inhibition of receptors on afferent nerves initiating reflexes that both accelerate and delay transit.

Short chain fatty acids in the terminal ileum accelerate stomach to caecum transit time in the rat.

We have previously shown that infusion of triglycerides and long chain fatty acids into the ileum of humans and rats delays small bowel transit time. The present studies have investigated the effect on the stomach to caecum transit time of a baked bean meal of the ileal infusion of 20 mM, 50 mM, and 100 mM acetic, butyric, hexenoic, and caprylic acids in rats. After an 18 hour fast either a control or a short chain fatty acid (SCFA) solution (pH 6.5) was infused into the ileum for 30 minutes (0.3 ml/hour). A test meal was given by gavage and the infusion continued for a further 150 minutes. The arrival of the meal in the colon was signalled by a rise in the exhaled hydrogen concentration. Acetic acid (20 mM, 50 mM, 100 mM), butyric acid (100 mM), and caprylic acid (100 mM) produced a significant acceleration of transit which was inversely proportional to SCFA chain length. In a separate experiment, infusion of 100 mM acetic acid, the most potent SCFA, into an isolated ileal Thirty-Vella loop failed to accelerate transit of the test meal. Our results suggest that SCFAs accelerate transit via a local enteric reflex.

The effect of ethanol on the normal food intake and eating behaviour of the rat.

A computerised system was used to continually monitor the food intake and meal patterns of six rats over eighteen days. The effect of replacing the normal drinking water with a 10% ethanol solution for the second six-day period was assessed. Fluid intake decreased by 29% and resulted in a mean ethanol consumption of 6 ml/kg/rat/day. Energy intake remained unchanged with food intake being significantly reduced (p less than 0.05) in proportion to the calories consumed as ethanol. The reduction in food intake was achieved by a significant decrease in mean meal size (p less than 0.01), whilst meal frequency demonstrated a nonsignificant increase. Withdrawal of the ethanol solution resulted in highly variable feeding behaviour. It is concluded that rats will adapt their meal patterns to regulate their energy intake when a 10% ethanol solution is the sole fluid source. It is suggested that the caloric potential of ethanol is recognised through the release of CCK and consequently meal pattern adaptations are the result of premature satiety.

Characteristics of lipid substances activating the ileal brake in the rat.

Studies were carried out in 36 adult male rats to determine the characteristics of lipid substances which, after infusion into the ileum, slow the stomach to caecum transit time of the head of a bean meal in the rat. Stomach to caecum transit time was measured by environmental hydrogen analysis. Ileal infusion of a range of free fatty acids including petroselinic, oleic, myristoleic, erucic, linoleic, and linolenic all significantly slowed stomach to caecum transit time, as did the detergents (sodium bis (2-ethyl hexyl) sulphosuccinate and sodium linoleyl sulphate), the triglyceride corn oil, and the phospholipid lecithin. Although the lipid soluble deconjugated bile acid deoxycholic acid slowed stomach to caecum transit time, the water soluble conjugated bile acid taurocholic acid accelerated it. Infusion of the lipid alcohol oleyl alcohol and the calcium chelating agent disodium edetate (EDTA) into the ileum did not delay the passage of the meal through the stomach and small intestine. The diversity of lipid substances activating the ileal brake suggest a nonspecific effect by lipid soluble substances that can penetrate cell membranes. The lack of effect of EDTA suggested that calcium binding was not important.

The effect of acute and chronic administration of morphine and morphine withdrawal on intestinal transit time in the rat.

The effects of acute and chronic morphine administration and of morphine-withdrawal on intestinal transit time of a liquid meal were investigated using rats. Many experiments have assessed the effects of acute morphine administration on intestinal transit, but the intestinal effects of chronic morphine administration have been neglected. Our results showed no significant differences between morphine-dependent and control animals when assessing the leading edge of the liquid meal infusion, its distribution and geometric centre (G.C.). However, during naloxone-precipitated withdrawal from morphine, the leading edge of the infusion and its G.C. were significantly distal to values obtained from other groups. Acute morphine administration caused delayed intestinal transit of a meal infusion, an effect partly caused by significant retention of the infusion in the stomach and duodenum. The leading edge of the meal infusion and G.C. were significantly proximal to values obtained from other groups of animals. The results show that morphine-dependent rats develop complete tolerance to the delayed intestinal transit of a meal observed after acute morphine administration and that withdrawal from morphine accelerates intestinal transit of a liquid meal.

The effect of guar gum on the distribution of a radiolabelled meal in the gastrointestinal tract of the rat.

1. The effect of addition of guar gum (5 and 10 g/l) to a radiolabelled, homogenized, baked-bean test meal on the distribution of that meal in the gastrointestinal tract was investigated in groups of male rats killed at 25, 50, 100, 200, 300 and 400 min after gavage. 2. Addition of 5 and 10 g guar gum/l significantly increased the proportion of the meal remaining in the stomach at 25 and 50 min after gavage (P less than 0.01). 3. The heads of the control meal and meals containing guar gum reached the distal intestine within 25 min after gavage but radioactivity was not observed in the caecum until 100 min after administration of each of the meals. Addition of guar gum (5 and 10 g/l) delayed caecal filling even though the head of each meal reached the caecum at the same time after gavage. 4. The geometric centres of guar-gum-containing meals were proximal to that of the control meal at all times after gavage. 5. The observed delay in the passage of a guar-gum-containing meal through the stomach and small intestine is probably due to the viscous nature of the meal resisting the propulsive and mixing effects of the gastrointestinal contractions, thereby reducing access of the glucose to the absorptive epithelium. This could contribute to the observed reductions in postprandial glycaemia seen in previous studies after incorporating guar gum into a meal.