Dietary fructose during the suckling period increases body weight and fatty acid uptake into skeletal muscle in adult rats

Objective: The suckling period is one potentially “critical” period during which nutritional intake may permanently “program” metabolism to promote increased adult body weight and insulin resistance in later life. This study determined whether fructose introduced during the suckling period altered body weight and induced changes in fatty acid transport leading to insulin resistance in adulthood in rats. Methods and Procedures: Pups were randomly assigned to one of four diets: suckle controls (SCs), rat milk substitute formula (Rat Milk Substitute), fructose‐containing formula (Fructose), or galactose‐containing formula (Galactose). Starting at weaning, all pups received the same diet; at 8 weeks of age, half of the SC rats began ingesting a diet containing 65% kcal fructose (SC‐Fructose). This continued until animals were 12 weeks old and the study ended. Results: At weeks 8, 10, and 11, the Fructose group weighed more than SC and SC‐Fructose groups (P < 0.05). At weeks 8 and 10 of age, the Fructose group had significantly higher insulin concentrations vs. rats in the SC‐Fructose group. ³H‐Palmitate transport into vesicles from hind limb skeletal muscle was higher in Fructose vs. SC rats (P < 0.05). CD36 expression was increased in the sarcolemma but not in whole tissue homogenates from skeletal muscle from Fructose rats (P < 0.05) suggesting a redistribution of this protein associated with fatty acid uptake across the plasma membrane. This change in subcellular localization of CD36 is associated with insulin resistance in muscle. Discussion: Consuming fructose during suckling may result in lifelong changes in body weight, insulin secretion, and fatty acid transport involving CD36 in muscle and ultimately promote insulin resistance.     

Age-associated changes in intestinal fructose uptake are not explained by alterations in the abundance of GLUT5 or GLUT2

A reduction in nutrient absorption may contribute to malnourishment in the elderly. The objectives of this study were to determine the effects of aging on the absorption of fructose in rats, as well as the mechanisms of these adaptive changes. Male Fischer 344 rats aged 1, 9, and 24 months were fed standard Purina chow for 2 weeks (PMI #5001, PMI Nutritionals, Brentwood, MO). The uptake of (14)C-labeled D-fructose was determined in vitro using the intestinal sheet method. Intestinal samples were taken for RNA isolation and for brush border membrane (BBM) and basolateral membrane (BLM) preparation. Northern blotting, Western blotting, and immunohistochemistry were used to determine the effects of age and diet on GLUT5 and GLUT2. When expressed on the basis of intestinal or mucosal weights, aging was associated with a decline in jejunal and ileal fructose uptake, whereas jejunal fructose uptake was increased when expressed on the basis of serosal or mucosal surface area. The alterations in fructose uptake were not paralleled by changes in GLUT5 or GLUT2 abundance. These results indicate that 1) the effect of age on fructose uptake depends on the method used to express results, and 2) the age-associated changes in uptake are not explained by alterations in GLUT5 and GLUT2.     

Dietary fructose vs glucose does not influence iron status in rats

The effect of dietary fructose vs glucose on iron status was studied in rats. Female rats were fed for 4 wk diets containing either fructose or glucose (709.4 g monosaccharide/kg). Fructose vs glucose lowered iron concentrations in liver, kidney, and heart, but did not alter absolute iron contents.     

Determination of unidirectional uptake rates for lipids across the intestinal brush border

An in vitro method is presented which measures valid, unidirectional uptake rates for lipids across the intestinal brush border. This method combines analysis by a newly devised, double isotope counting system for solubilized tissue with the use of a nonabsorbable marker to correct gross uptake determinations for contamination by adherent mucosal fluid. Of seven markers, only [(3)H]inulin measured adherent mucosal fluid volumes as much as 20% greater than the other markers. Diffusion of the nonabsorbable marker, as well as of the compound being studied, into the unstirred layer made the time course of uptake critically important. The time lag for diffusion of marker invalidates the use of 1-min incubation periods; however, a linear time course of uptake that intercepts essentially at zero was found for taurocholate and octanoate for periods of from 2 to 5 min. Working within this critical time period with jejunum, it was shown that tissue dry weight was an appropriate measure of the amount of tissue and that uptake rates for taurocholic, octanoic, and lauric acids were linear with respect to concentration. Tissue binding of compounds was not significant. The results demonstrate that careful use of the described method yields accurate measurement of unidirectional uptake rates of lipids across the brush border that are of critical importance in defining the characteristics of membrane penetration and the rate-limiting steps in fat and sterol absorption.     

Dietary fructose vs glucose lowers copper solubility in the digesta in the small intestine of rats

The hypothesis was tested that dietary fructose vs glucose lowers copper solubility in the digesta in the small intestine of rats, which in turn causes a decreased copper absorption. Male rats were fed adequate-copper (5 mg Cu/kg) diets containing either fructose or glucose (709.4 g monosaccharide/kg) for a period of 5 wk. Fructose vs glucose significantly lowered copper concentrations in plasma and the liver, but did not alter hepatic copper mass. Fructose feeding resulted in a significantly lesser intestinal solubility of copper as based on either a smaller soluble fraction of copper in the liquid phase of small intestinal contents or a lower copper concentration in the liquid phase. The latter fructose effect can be explained by the observed fructose-induced increase in volume of liquid phase of intestinal digesta. After administration of a restricted amount of diet extrinsically labeled with⁶⁴Cu, rats fed fructose also had significantly lower soluble⁶⁴Cu fraction in the digesta of the small intestine. Although this study shows that fructose lowered intestinal copper solubility, only a slight reduction of apparent copper absorption was observed. It is suggested that the fructose-induced lowering of copper status in part counteracted the fructose effect on copper absorption at the level of the intestinal lumen.     

Analysis of age-associated changes in collagen crosslinking in the skin and lung in monkeys and rats

The present study was designed to address a specific question: can we define collagen aging in vivo in terms of alterations in collagen crosslinking? In order to assess the complete spectrum of change throughout life, tissues from rats, monkeys and (where available) humans were examined at ages ranging from fetal to old. Skin and lung were selected in order to include all of the crosslinks derived from lysyl oxidase-generated aldehydes that have been identified thus far, both reducible and nonreducible. Crosslinks analyzed included hydroxylysinonorleucine, dihydroxylysinorleucine, histidinohydroxymerodesmosine, hydroxypyridinium, lysyl pyridinium, and a deoxy analogue of hydroxypyridinium found in skin that differs structurally from lysyl pyridinium. Tissues from both a short-lived species (rats) and a long-lived species (monkeys) were analyzed to test further the hypothesis that changes in crosslinking are linked predominantly to biological age of the animal, rather than temporal aging. We found that biological aging seems to regulate certain predictable changes during the first part of the lifespan: the disappearance postnatally of dihydroxylysinonorleucine in skin, the rapid decrease in difunctional crosslink content in lung and skin during early growth and development, and the gradual rise in hydroxypyridinium and lysyl pyridinium in lung tissue. Changes in crosslinking were far less predictable during the second half of the lifespan. Although hydroxypridinium content continued to rise or reached a plateau in rat and monkey lungs, respectively, it showed a decrease in human lungs. The analogous trifunctional crosslink in skin, the so-called 'pyridinoline analogue', decreased dramatically in both rats and monkeys in later life. Our data suggest that caution must be taken in drawing inferences about human connective tissue aging from experiments performed in short-lived species such as rodents. Furthermore, the finding that there may be fewer total lysyl oxidase-derived crosslinks per collagen molecule in very old animals as compared with young animals suggests that we may need to expand our concepts of collagen crosslinking.     

Dietary fructose and intestinal barrier: potential risk factor in the pathogenesis of nonalcoholic fatty liver disease

Worldwide, not only the prevalence of obesity has increased dramatically throughout the last three decades but also the incidences of co-morbid conditions such as diabetes type 2 and liver disease have increased. The ‘hepatic manifestation of the metabolic syndrome’ is called nonalcoholic fatty liver disease (NAFLD) and comprises a wide spectrum of stages of liver disease ranging from simple steatosis to liver cirrhosis. NAFLD of different stages is found in ～30% of adults and ～20% in the US population. Not just a general overnutrition but also an elevated intake of certain macronutrients such as fat and carbohydrates and herein particularly fructose has been claimed to be risk factors for the development for NAFLD; however, the etiology of this disease is still unknown. The present review outlines some of the potential mechanisms associated with the development of NAFLD and fructose intake with a particular focus on the role of the intestinal barrier functions.     

Locally and systemically active glucocorticosteroids modify intestinal absorption of sugars in rats.

Glucocorticosteroids enhance digestive and absorptive functions of the intestine of weaning and adult rats. This study was undertaken to assess the influence of treatment of weaning male rats with budesonide (Bud), prednisone (Pred), or control vehicle on the in vitro jejunal and ileal uptake of glucose and fructose. Bud and Pred had no effect on the uptake of d-glucose by sodium glucose transporter-1. In contrast, the uptake of d-fructose by GLUT-5 was similarly increased with Bud and with Pred. The increases in the uptake of fructose were not due to variations in the weight of the intestinal mucosa, food intake, or in GLUT-5 protein or mRNA expression. There were no steroid-associated changes in mRNA expression of c-myc, c-jun, c-fos, proglucagon, or selected cytokines. However, the abundance of ileal ornithine decarboxylase mRNA was increased with Pred. Giving postweaning rats 4 wk of Bud or Pred in doses equivalent to those used in clinical practice increases fructose but not glucose uptake. This enhanced uptake of fructose was likely regulated by posttranslational processes.     

Dietary polyphenols decrease glucose uptake by human intestinal Caco-2 cells

The effect of different classes of dietary polyphenols on intestinal glucose uptake was investigated using polarised Caco-2 intestinal cells. Glucose uptake into cells under sodium-dependent conditions was inhibited by flavonoid glycosides and non-glycosylated polyphenols whereas aglycones and phenolic acids were without effect. Under sodium-free conditions, aglycones and non-glycosylated polyphenols inhibited glucose uptake whereas glycosides and phenolic acids were ineffective. These data suggest that aglycones inhibit facilitated glucose uptake whereas glycosides inhibit the active transport of glucose. The non-glycosylated dietary polyphenols appear to exert their effects via steric hindrance, and (-)-epigallochatechingallate, (-)-epichatechingallate and (-)-epigallochatechin are effective against both transporters.     

D-glucose uptake in intestinal brush-border membrane vesicles of rachitic rats

We have previously reported the metabolic consequences of feeding rats Steenbock and Black's rickets-inducing diet, deficient in vitamin D and with an altered Ca/P ratio. Using isolated brush-border membrane vesicles prepared from the jejunum, ileum and duodenum of control and rachitic rats, we have demonstrated a marked decrease of Na+-dependent D-glucose uptake at jejunum-ileum level of rachitic rats. At duodenum level Na+-dependent D-glucose transport was not influenced by rickets. A lack of any significant difference between the two animal groups was observed studying the facilitated transport of D-glucose, the diffusion of L-glucose and the Na+-dependent uptake of phenylalanine and aspartate.     

Dietary hexachlorocyclohexane induced changes in blood and liver lipids in albino mice.

Dietary intake of technical hexachlorocyclohexane (HCH) by albino mice for 2 weeks (at 400 and 800 ppm) resulted in hyperlipemia. Significant increase in triglycerides, phospholipids and cholesterol fractions of blood was observed in these animals. Dietary intake of gamma-isomer of HCH for 2 weeks (at 200 ppm) did not have any effect on blood lipid profile, but at 400 ppm level produced higher contents of phospholipids and cholesterol. The hepatomegaly produced by dietary technical HCH or gamma-HCH was not accompanied by fatty metamorphosis of liver. Hypertriglyceridemia caused by HCH was accompanied by lower triglyceride levels in liver, suggesting a possible higher rate of secretion from liver.     

Diurnal changes in liver and plasma lipids of choline-deficient rats

Early effects of choline deficiency were studied in rats. Nonphospholipid ("neutral lipid") and phospholipid were measured in plasma and in three fractions of a liver homogenate: sediment, supernatant fraction, and "floating fat." A single choline-deficient meal caused significant aberrations from the typical diurnal changes observed in the lipid fractions of the controls. These changes occurred in the following sequence: (a) failure of phospholipid to increase, after feeding, in the sediment fraction; (b) increase of neutral lipid, compared with controls, exclusively in the floating fraction; and (c) failure of neutral lipid to return to control levels. The rate of accumulation of neutral lipid increased during the first 4 days of deficiency. The occurrence of NADH-cytochrome c dehydrogenase in the floating fat and the absence of succinate dehydrogenase activity point to microsomal origin of the floating fat. Early effects of choline deficiency on plasma lipids were limited to phospholipid, and occurred later than changes in the liver. Plasma nonphospholipid levels were unchanged during the first 2 days; this does not support impaired secretion or transportation of glyceride as the cause of fatty liver in the early stages of choline deficiency.     

Chronic changes in plasma triglyceride levels do modify platelet membrane microviscosity in rats

Lipid metabolism disorders were proposed to mediate numerous cell membrane alterations in various forms of hypertension. Elevated plasma triglycerides were found to be associated with changes in membrane structure and function related to altered microviscosity in particular domains of the cell membrane. The aim of our study was to determine if an abnormal triglyceride metabolism might play a causal role in these alterations of membrane dynamics. Using genetically hypertensive rats of the Prague hereditary hypertriglyceridemic (HTG) strain we investigated whether the elevation of circulating triglycerides induced by high fructose intake and/or their lowering by chronic gemfibrozil treatment (for 10 weeks starting at the age of 6 weeks) are followed by reciprocal changes in membrane microviscosity. Two different fluorescent probes exploring either the outer membrane leaflet (TMA-DPH anisotropy) or the membrane lipid core (DPH anisotropy) were used in platelets of HTG rats. DPH (diphenylhexatriene) fluorescence anisotropy was decreased in platelets of fructose-treated HTG animals with highly elevated plasma triglyceride levels, whereas it was increased in gemfibrozil-treated HTG rats in which triglyceride levels were almost normalized. On the contrary, TMA-DPH (trimethylamino-diphenylhexatriene) anisotropy was not substantially altered in platelets from HTG rats by the above modifications of circulating triglycerides. No changes of plasma cholesterol or blood pressure were associated with the triglyceride-dependent modifications of membrane core microviscosity. Our interventional study demonstrates a major causal role of circulating triglycerides in the control of the microviscosity of membrane lipid core.     

Seasonal changes in plasma and aortic lipids in young rats

In young non-exercised rats, plasma triglyceride and plasma phospholipid levels increased in summer and low density lipoprotein cholesterol (LDL-C) increased in winter. As for lipids samples from the wall of the aorta, total cholesterol, cholesteryl ester and triglyceride decreased in summer and phospholipid decreased in winter. Exercise diminished the gain in body mass (delta m) and suppressed seasonal changes in the levels of LDL-C and plasma triglyceride. Seasonal changes in the aorta lipids in this case were similar to those found in non-exercised animals. The values of total energy intake (Q) and of delta m.Q-1 were found to change with season in both non-exercised and exercised rats. Seasonal changes in plasma and in aorta lipids observed in these animals ran in parallel with the respective levels of delta m.Q-1 and/or of Q. The training effect on the lipid values detected in summer and/or in winter was also found to be dependent on the reduction in delta m.Q-1 with exercise. In the non-exercised and in the exercised animals, plasma phospholipid was associated with aorta phospholipid and inversely related to aorta cholesteryl ester and aorta triglyceride. The relationship between these estimations suggests that an increase in the plasma phospholipid in summer would remove non-polar lipids from the walls of the aortae.     

Suppressive effect of nobiletin and epicatechin gallate on fructose uptake in human intestinal epithelial Caco-2 cells

Abstract

Inhibition of excessive fructose intake in the small intestine could alleviate fructose-induced diseases such as hypertension and non-alcoholic fatty liver disease. We examined the effect of phytochemicals on fructose uptake using human intestinal epithelial-like Caco-2 cells which express the fructose transporter, GLUT5. Among 35 phytochemicals tested, five, including nobiletin and epicatechin gallate (ECg), markedly inhibited fructose uptake. Nobiletin and ECg also inhibited the uptake of glucose but not of L-leucine or Gly-Sar, suggesting an inhibitory effect specific to monosaccharide transporters. Kinetic analysis further suggested that this reduction in fructose uptake was associated with a decrease in the apparent number of cell-surface GLUT5 molecules, and not with a change in the affinity of GLUT5 for fructose. Lastly, nobiletin and ECg suppressed the permeation of fructose across Caco-2 cell monolayers. These findings suggest that nobiletin and ECg are good candidates for preventing diseases caused by excessive fructose intake.     

Developmental changes in intestinal globule leukocytes of normal rats

Among vertebrates, telencephalo-pontine systems exist only in birds and mammals. However, three nuclei in the diencephalon and mesencephalon of teleost fishes have been indicated — analogous to the pons — to represent relay stations between telencephalon and cerebellum. Since two of these nuclei (dorsal preglomerular nucleus, dorsal tegmental nucleus) have only been described in the highly derived, electrosensory mormyrids, we investigated telencephalic connections in two nonelectrosensory teleosts, the goldfish Carassius auratus and the freshwater butterflyfish Pantodon buchholzi, and cerebellar connections only in the latter species, since for C. auratus these connections are already established. Horseradish peroxidase tracing reveals that C. auratus has a dorsal tegmental nucleus and a paracommissural nucleus both of which are telencephalo-recipient and project to the cerebellum, and that P. buchholzi has a dorsal preglomerular nucleus with such connections. These results extend our knowlegde of the distribution and, therefore, the phylogeny of telencephalo-cerebellar systems in teleosts. Similar to tetrapods, teleosts appear to have developed telencephalo-cerebellar systems several times independently.