Jennifer M. Gee

Quercetin Glucosides Interact With the Intestinal Glucose Transport Pathway

Flavonols are efficient antioxidants with the potential to protect biological macromolecules from oxidative damage in vivo, and if absorbed into the circulation they may protect against cardiovascular disease. Although flavonol aglycones are present in foods at low concentrations, their glycosides are abundant in onions, apples, beans and tea, and are thought to be stable under the conditions of the human stomach and small bowel. There is, however, recent evidence to suggest that intact glycosides of quercetin may be absorbed from the small intestine by a mechanism involving the glucose transport pathway. In the present study we tested this hypothesis by measuring the effect of quercetin glycosides on the rate of efflux of galactose from the jejunal mucosa. Everted sacs of rat jejunum preloaded with 14C-galactose were exposed to quercetin glycosides isolated from onions. Quercetin mono- and diglucosides were shown to accelerate the carrier-mediated efflux of galactose via a sodium-dependent pathway. HPLC analysis confirmed the stability of the glycosides under conditions simulating those in the upper alimentary tract. These studies suggest that purified quercetin glucosides are capable of interacting with the sodium dependent glucose transport receptors in the mucosal epithelium and may therefore be absorbed by the small intestine in vivo.

Effect of oat gum on the physical properties of the gastrointestinal contents and on the uptake of D-galactose and cholesterol by rat small intestine in vitro

Recent reports indicate that oats have a relatively low glycaemic effect in comparison with other carbohydrate food, and that their consumption leads to a reduction in plasma-cholesterol levels in man. These properties may be due to a soluble non-starch polysaccharide in oats. The present study was undertaken to explore the physiological properties of this material. Three groups of male Wistar rats were meal-fed on a control diet free of soluble dietary fibre for 10 d before being given a 10 g meal of either the control diet, a diet containing oat gum (beta-glucan), or finely ground rolled oats. The contents of the stomach, small intestine and caecum were later recovered and the weight, water content and viscosity were measured. The small intestinal contents from oat-gum-fed or oat-fed rats had a higher wet: dry weight ratio than that of the controls, and a higher viscosity. In in vitro studies the rate of uptake of D-galactose by jejunal rings was reduced in the presence of oat gum. The estimated Michaelis-Menten constant for the carrier-mediated component in the presence of oat gum was higher than that for controls, but the maximum transport rates were similar. Cholesterol uptake by everted jejunal sacs was progressively inhibited by increasing concentrations of oat gum in the mucosal medium. It is concluded that increased viscosity of the contents of the small intestine may contribute to the low glycaemic index and hypocholesterolaemic effects of oats in man. Oats appear to be amongst the few palatable sources of viscous dietary fibre in the conventional Western diet.

Gastrointestinal adaptation in response to soluble non-available polysaccharides in the rat

1. Rats were fed on a control semi-synthetic diet containing insoluble cellulose (Solkafloc; 100 g/kg; control group) as the only source of dietary fibre, or on one of two test diets containing the same quantity of either guar gum or carboxymethylcellulose (CMC). Animals in the test groups showed similar growth rates and food intakes, which were significantly lower than those of the control group. The CMC group produced frequent poorly formed faeces throughout the 21 d feeding period. 2. The small intestines of animals in both test groups were significantly longer than those of the control group at the end of the study. The caeca were also enlarged and heavier, particularly in the CMC-fed group. 3. The rate of production of mucosal cells was increased in the small and large intestines of both test groups. The CMC-fed group exhibited a particularly high rate in the distal ileum, where the rate of cell divisions per crypt was over three times greater than at the same site in the control group. The increased proliferation was associated with a significant lengthening of the crypts and an approximately 25% increase in the basal width of the villi. 4. Mucosal alkaline phosphatase (EC 3.1.3.1) and lactase (EC 3.2.1.23) levels were lower than those of the control group at proximal and distal sites in the small intestines of both CMC- and guar-gum-fed groups. Altered spatial distributions of maltase (EC 3.2.1.20) and sucrase (EC 3.2.1.48) activities were also observed in these animals.(ABSTRACT TRUNCATED AT 250 WORDS)

The influence of guar gum on intestinal cholesterol transport in the rat

Everted sacs of rat proximal small intestine were used to determine the effect of guar gum (5 g/l) on the uptake of cholesterol (0.1 mM) from a solution of micelles. The uptake of cholesterol was found to be linear both in the presence and absence of guar gum. When guar was present throughout the whole of the incubation medium, the uptake of cholesterol was reduced to approximately 40% of control values. Sacs which had been pre-incubated in guar gum before exposure to cholesterol in a guar-free medium also showed a reduction in cholesterol uptake but this was less pronounced. A two-stage perfusion technique, previously described (Blackburn & Johnson, 1981), was used to determine the effect of a guar layer adsorbed to the mucosal surface on cholesterol absorption in vivo. Such a layer leads to a reduction of approximately 36%; it was concluded that guar slows the absorption of cholesterol from micelles by a mechanism, or mechanisms, involving an increased resistance to diffusion in the aqueous medium. Groups of rats were meal-fed for at least 30 d on semi-synthetic diets with or without the inclusion of guar gum (20 g/kg). Rates of intestinal absorption of cholesterol, glucose and fluid were then determined by the perfusion technique in vivo. There was no reduction in absorption in the test animals compared with the controls. It is proposed that guar gum is able to slow the intestinal transport of cholesterol from a suspension of pre-formed micelles, but only when both are present in the lumen together. No evidence was obtained to suggest that the consumption by rats of a diet containing guar gum, at a level similar to that used in human studies, leads to any adaptive reduction in their rates of cholesterol or glucose absorption.

Effect of dietary supplements of guar gum and cellulose on intestinal cell proliferation, enzyme levels and sugar transport in the rat.

Male Wistar rats (approximately 200 g) were given fibre-free semi-synthetic diets containing either sucrose (S) or a sucrose-starch mixture (SS) as the carbohydrate component, or a diet similar to SS containing 40 g guar gum/kg (G), or 100 g cellulose/kg (C). The animals remained healthy, and weight gain after 30 d was similar in all groups. The small intestines of the animals given diet G were significantly longer than those of the other groups, and showed signs of increased mitotic activity and mucosal growth. No significant differences in mucosal enzyme activity were detected between the two fibre-free control groups. Lactase (EC 3.2.1.23) and alkaline phosphatase (EC 3.1.3.1) activities were significantly lower than controls in group G, but were higher in group C. Kinetic analysis of 3-O-methyl glucose uptake by isolated intestine indicated that the maximum transport rate (Vmax) of tissue from group G tended to be lower than from the fibre-free group SS and group C. It is concluded that materials which are classed as dietary fibre but which differ markedly in their physical properties may also differ in the functional changes to which they give rise in the small intestine. These changes may be at least partially mediated by effects on mucosal cell proliferation.

Intestinal microflora and gastrointestinal adaptation in the rat in response to non-digestible dietary polysaccharides

1. A comparison was made of the effect of a fibre-free diet and diets containing non-digestible polysaccharides on rat caecal and colonic physiology and microflora. 2. All polysaccharide-containing diets led to enlargement of the caecum and colon, associated with increased weight of contents, and of tissue. Carboxymethylcellulose (CMC) had the most marked effect and animals given this also had watery faeces. 3. The density of bacteria in the caecum and colon varied significantly with diet and the proportion of aerobic bacteria in the flora was increased by the CMC diet. 4. In vitro, CMC and hydroxypropylmethylcellulose were poorly fermented. 5. There was a high correlation (caecum r 0.93; colon r 0.94) between tissue weight and wet weight of organ contents but no correlation with bacterial density, number of bacteria per organ, moisture content or short-chain fatty acid content. 6. It is concluded that caecal and colonic enlargement is due to tissue hypertrophy in response to increased bulk of contents, irrespective of the nature of that bulk which varies with diet; it is unlikely that short-chain fatty acids or other microbial metabolites are the stimulus for the trophic response seen when non-digestible dietary polysaccharides are fed to rats.

Differences in intestinal protein synthesis and cellular proliferation in well-nourished rats consuming conventional laboratory diets

Male Wistar rats (100 g) were given a commercial pellet feed or a semi-synthetic diet ad lib. Although the pellet-fed group grew slightly faster than the other group during the early part of the feeding period, there was no significantly difference between the final weights of the groups. The fractional rates of protein synthesis in jejunum, proximal ileum and liver were measured by a technique based on the determination of L-[4-3H]phenylalanine incorporation over a short time period. Protein synthesis was higher in both jejunum and ileum of the pellet-fed rats compared with those eating the semi-synthetic diet, but there was no difference between the rates of protein synthesis measured in the livers of the groups. The rate of mucosal cell division was significantly faster in the ileal mucosa of the pellet-fed group compared with the other group, and there were significant differences in some aspects of mucosal morphology. The maintenance of higher rates of cell turnover and protein synthesis in animals given a commercial pellet feed is unexplained, but it may be related to the presence of non-absorbable polysaccharides or other complex plant materials in the pellet feed.

The effect of Gypsophila saponins in the diet on mineral status and plasma cholesterol concentration in the rat

1. Immature, male Wistar rats were allocated to one of six groups and caged individually. The first group was given a semi-synthetic diet containing 38 mg iron and 55 mg zinc/kg (basal group). The second and third groups were given a diet containing 10 mg Zn and 12 mg Fe/kg respectively (low-Zn and low-Fe groups). Groups four, five and six were given similar diets containing 20 g Gypsophila saponins/kg. After 21 d the Fe and Zn status of the rats was estimated and plasma cholesterol concentration determined. 2. Measurements of whole blood haemoglobin concentration, packed cell volume and liver Fe stores indicated that rats in the basal + saponin and low-Fe + saponin groups had a significantly reduced Fe status when compared with their controls. Rats in the low-Zn + saponin group also showed a trend toward reduced Fe stores. 3. Zn status, as judged by femur Zn concentration, was not adversely affected by the inclusion of Gypsophila saponins in the diet. 4. Consumption of the saponins resulted in a significant reduction in blood cholesterol concentration, with rats in both the low-Fe groups having significantly lower concentrations than their basal and low-Zn counterparts. 5. In view of suggestions that the consumption of saponins should be encouraged because of their ability to lower blood cholesterol, possible effects on Fe metabolism should be investigated further, particularly with respect to the levels and sources of saponin in the human diet.

Intestinal cellular proliferation and protein synthesis in zinc-deficient rats

Immature male Wistar rats were given a low-zinc semi-synthetic diet (2 mg Zn/kg) for 28 d. Control groups received a similar diet supplemented with 58 mg Zn/kg either ad lib. or in amounts matched to the consumption of the Zn-deficient group. Rates of growth, food consumption and small intestinal length were significantly reduced in the Zn-depleted rats. Zn deficiency in the rat was associated with a reduction in the ratio, crypt: villus and a lower rate of crypt cell division in the jejunum. This resulted in a substantial decrease in the net influx of new cells into the villi of the Zn-deficient animals compared with controls. The fractional rates of protein synthesis in jejunal mucosa were measured by a technique based on the determination of L-[4-3H]phenylalanine incorporation. There was no evidence of a decline in the protein synthetic rate in total mucosa from Zn-deficient rats. It is suggested that a reduction in cell influx into the villi may be responsible for the morphological and functional changes observed in the small intestine of rats fed on a low-Zn diet.

Intestinal microflora, morphology and enzyme activity in zinc-deficient and Zn-supplemented rats

1. Immature, male Wistar rats were given a low-zinc diet (2 mg/kg) for 22-24 d. Control groups received a similar diet supplemented with 58 mg Zn/kg either ad lib., or in amounts matched to the consumption of the Zn-deficient group. Food consumption, rate of growth and food conversion efficiency were markedly lower in the Zn-deficient group of rats compared with controls. Appetite, growth rate and food utilization improved dramatically over a subsequent 4 d period of Zn supplementation. 2. Morphological examination of samples of jejunum and ileum confirmed that Zn deficiency in the rat is accompanied by a reduction in villous dimensions and increase in villous density. After a short period of Zn supplementation, villous density and the basal width and maximum height of individual villi in the jejunum returned to normal. Similar changes occurred in the ileum but to a lesser extent. 3. Mucosal alkaline phosphatase (EC 3.1.3.1) activity was significantly lower in the small intestine of Zn-deficient rats compared with Zn-supplemented rats. Disaccharidase activities were lower in the Zn-deficient group, compared with their feed-restricted counterparts, but were similar to values for ad lib.-fed controls. Tissue alkaline phosphatase and disaccharidase activities were consistently higher after a 4 d period of Zn supplementation, compared with non-supplemented animals, but this increase was only significant for alkaline phosphatase. 4. Although there were striking similarities in the mucosal characteristics of gnotobiotic and Zn-deficient rats, there was no indication that even severe dietary Zn depletion reduced the numbers of viable bacteria present in either the small or large intestine.(ABSTRACT TRUNCATED AT 250 WORDS)