Studies on the development of an insulin resistant rat model by chronic feeding of low magnesium high sucrose diet.

Magnesium deficiency and excess sucrose in the diet have been shown to play an important role in the development of insulin resistance. In the present study we have looked at the combined effect of a low magnesium high sucrose diet on basal glucose and insulin levels, erythrocyte insulin receptors and lipid profile in rats. For this purpose rats were divided into four groups and fed control, low magnesium, high sucrose and low magnesium high sucrose diets respectively for three months. The biochemical analysis showed a significant increase in blood glucose and triglyceride levels after one, two and three months of feeding in both the high sucrose and the low magnesium high sucrose groups, while rats fed a low magnesium diet showed a significant increase in blood glucose and triglyceride levels only after the second month. Insulin levels increased significantly in low magnesium, high sucrose and low magnesium high sucrose groups by the end of the study period. Compared to control rats, the binding of insulin to the erythrocyte insulin receptors was reduced significantly in the high sucrose and the low magnesium high sucrose groups. Cholesterol levels were found to increase significantly in the high sucrose group at the end of one month and three months of feeding. HDL-cholesterol decreased significantly in the low magnesium high sucrose group by the end of the study. Serum and RBC magnesium levels demonstrated a significant decrease in the low magnesium and the low magnesium high sucrose groups. The post heparin plasma lipoprotein lipase activity was decreased significantly in low magnesium, high sucrose and low magnesium high sucrose groups compared to control rats. These findings suggest that feeding a diet low in magnesium and high in sucrose causes insulin resistance in rats.     

Overeating, overweight and obesity induced by an unpreferred diet

Rats fed diets containing 50-71% added water (liquid diets) eat more energy and gain more weight than rats fed the same diets without added water (solid diets). The present experiments examined the effects of making a liquid diet less palatable. The first experiment examined the effects of sucrose octaacetate on diet preference. Rats, given a choice of a liquid diet containing 0.5% sucrose octaacetate and a plain solid diet, preferred the plain solid diet for three weeks. When the concentration of sucrose octaacetate was reduced to 0.05%, the rats did not show a reliable preference for either the sucrose octaacetate liquid or plain dry diet. In subsequent experiments, each rat was given only one diet at a time. In the second experiment, rats were fed 0.5% sucrose octaacetate liquid diet for three weeks followed by 0.05% sucrose octaacetate liquid diet for another four weeks. The rats fed the sucrose octaacetate liquid diet overate and became obese compared to the rats fed plain solid diet throughout. In the third experiment, rats fed 0.5% sucrose octaacetate liquid diet for six weeks became obese compared to rats fed plain solid diet throughout. Thus, the overeating and obesity induced by liquid diets cannot be attributed solely to their high palatability.     

Non-cariogenicity of the disaccharide palatinose in experimental dental caries of rats.

The caries-inducing activity of palatinose (isomaltulose, alpha-D-glucopyranosyl-1,6-fructose) was examined in in vitro and in vivo experiments, comparing it with other carbohydrates. When Streptococcus mutans was successively subcultured in a broth medium containing 1% palatinose, the strains belonging to serotype a, d, or g did not ferment palatinose, whereas the strains belonging to serotype b, c, e, or f did ferment palatinose. Furthermore, palatinose significantly inhibited the synthesis of insoluble glucan from sucrose by S. mutans. Specific-pathogen-free rats which had been infected with S. mutans 6715 and fed a diet containing 56% palatinose did not develop significant dental caries. However, rats infected similarly, but fed a diet containing sucrose, glucose, fructose, or a glucose-fructose mixture manifested significant caries when compared with the noninfected, sucrose-fed control rats. Furthermore, it was found that replacement of half of the sucrose content with palatinose resulted in decreased caries development compared with caries development in rats fed the sucrose diet.     

Sucrose-induced hyperphagia and obesity in rats fed a macronutrient self-selection diet

Adult female rats were allowed to self-select their diet from separate sources of fat, protein, and carbohydrate (starch). Other rats were fed a composite diet that matched the nutrient composition chosen by the self-selecting rats (50% fat, 28% protein, 22% carbohydrate) or a low-fat, high-carbohydrate chow diet. Half of the rats in each diet condition were given access to a 32% sucrose solution for 30 days. Sucrose availability increased total caloric intake (approximately 20%) and body weight gain in all three groups compared to control groups not fed the sucrose solution. The selection animals compensated for their sucrose intake by reducing their fat intake, and to a lesser degree, their starch intake; protein intake was the least affected by sucrose availability. The selection rats consumed less sucrose than the chow-fed rats and displayed a smaller increase in weight, relative to controls, than the chow-fed rats. These differences were attributed to the high-fat intake of the selection animals since similar results were obtained with the rats fed the composite diet. In particular, both the selection and composite diets produced mild obesity in the absence of sucrose. The results demonstrate that sucrose-induced overeating and overweight is not an artifact of restraining the diet choices of rats to a pure sugar and a nutritionally complete diet.     

Sex differences in the effects of voluntary activity on sucrose-induced obesity

Sedentary, adult rats of both sexes fed Purina chow and a 32% sucrose solution overate, gained excess weight and had higher Lee Indexes of obesity than control animals fed only Purina chow. The magnitude of these effects was similar in the males and females. Animals of both sexes fed the sucrose diet showed a slower rate of weight loss during food deprivation than the chow controls. Access to an activity wheel led to a reduction in caloric intake and the elimination of obesity in male rats. In the chow fed male rats activity led to a smaller, transient suppression in caloric intake and a slightly lower level of body weight than the sedentary chow controls. Access to activity did not affect body weight in the female rats in either dietary condition. Rather, both active groups of female rats appeared to compensate for the energy cost of voluntary activity by a small increase in food consumption. Long-term exposure to activity was associated with more rapid weight loss during food deprivation in both males and females. These data reveal that high levels of activity and obesity can co-exist when normal female rats are fed a palatable diet but that activity eliminates this form of obesity in the male rat.     

Accelerated Loss of Islet β Cells in Sucrose-Fed Goto-Kakizaki Rats, a Genetic Model of Non-Insulin-Dependent Diabetes Mellitus

The Goto-Kakizaki (GK) rat is a spontaneously diabetic animal model of non-insulin-dependent diabetes mellitus, which is characterized by progressive loss of β cells in the pancreatic islets with fibrosis. In the present study, we examined the effects of sucrose feeding on the islet pathology in this model. Six-week-old GK rats were fed with 30% sucrose for 6 weeks to induce severe hyperglycemia, and their condition was compared with that of nontreated rats. Age-matched normal Wistar rats were also given sucrose for the same periods and used for comparison. The sucrose-treated GK rats showed elevated blood glucose levels on oral glucose tolerance tests at 60 minutes and 120 minutes, representing 123% and 127% of values in untreated GK rats, respectively. At the end of the study, the mean β-cell volume density in GK rats was 50% less than that in untreated Wistar rats. Sucrose feeding further reduced the volume densities of β cells to only 50% of the levels of age-matched GK rats. Apoptotic cells were found in islet β cells only in GK rats fed sucrose (mean 0.067%). There appeared to be more islets that immunohistochemically stained strongly positive with 8-hydroxy-deoxyguanosine as a marker of oxidative damage of DNA in GK rats fed sucrose compared with those not given sucrose. GK rats not fed sucrose showed significantly lower proliferative activity of β cells measured by 5-bromo-2′-deoxyuridine uptake and intensified expression of Bcl-2 immunoreactivities at 6 weeks of age compared with those in age-matched Wistar rats. These two indices were reduced in both GK and Wistar rats with increasing age and were not affected by sucrose feeding in either group. The present study thus indicated that sucrose feeding promoted the apoptosis of β cells in GK rats through increased oxidative stress without altering their proliferative activity.     

Sucrose rich diet modifies thermogenic response to injection of muscimol into the posterior hypothalamus in the rat

The firing rate of the nerves innervating interscapular brown adipose tissue, temperatures of colon and interscapular brown adipose tissue, heart rate and oxygen consumption were monitored in urethane-anaesthetized male Sprague-Dawley rats fed with a sucrose rich diet. These variables were measured for 40 min before (baseline values) and 40 min after a 56 ng muscimol injection into the posterior hypothalamus. The same variables were monitored in other rats fed with a laboratory standard diet. Saline was injected into the posterior hypothalamus of control rats fed with sucrose or standard diet. Muscimol injection induced a decrease in firing rate, interscapular brown adipose tissue and colonic temperatures and oxygen consumption. This reduction was more evident in the rat fed with a sucrose rich diet than animals fed with standard diet. The kind of diet did not modify the decrease in heart rate induced by muscimol. These findings suggest that a sucrose rich diet modifies GABA-ergic responses to muscimol injection into the posterior hypothalamus.     

High fructose feeding of magnesium deficient rats is associated with increased plasma triglyceride concentration and increased oxidative stress.

The purpose of this study was to assess whether dietary carbohydrate could differentially influence the consequences of magnesium deficiency with particular emphasis on lipid metabolism and oxidative stress. Rats were fed a sucrose based or starch based diet either adequate or deficient in magnesium for two weeks. Magnesium deficient rats, as compared with rats fed magnesium adequate diets, displayed the usual decrease in plasma magnesium concentration. The classic symptoms of inflammation including hyperaemia, increased number of blood leukocytes and enlarged spleen weight were observed in these rats. Plasma TG and plasma apo B concentrations were also significantly increased. In addition, magnesium-deficient animals presented an increased susceptibility to lipid peroxidation of heart and liver tissues as shown by TBARS concentration. Regardless of magnesium status, sucrose feeding did not affect the magnesium plasma level and inflammatory parameters. Feeding rats the sucrose diets induced hypertriglyceridaemia and increased plasma apo B concentration. Heart and liver susceptibility to lipid peroxidation were significantly increased in rats fed the sucrose diets as compared with those fed the starch diets. Sucrose feeding in magnesium deficient rats was associated with higher plasma triglycerides concentration and higher tissue susceptibility to peroxidation as compared with magnesium deficient rats fed the starch diet. The results emphasised the potential detrimental and additional effect of sucrose feeding and magnesium deficiency on cardiovascular risk. Since the intake of magnesium has been reduced appreciably in industrialised countries while fructose consumption has been rapidly increased, the impact of this eating pattern should be clarified in humans.     

Functional alterations of liver mitochondria in chronic experimental alcoholism.

Mitochondria obtained from nonfatty livers of male rats fed an alcoholic “super diet” for 4 months displayed enlargement and bizarre configurations. In vitro, the mitochondria from alcohol-treated rats showed reduced oxidation of succinate and of malate-glutamate, but the energy coupling remained apparently unchanged when compared with that of three different controls. With β-hydroxybutyrate as substrate, the maximal respiratory rate was affected in relation to those of control rats in whose regimens alcohol was isocalorically substituted by sucrose or fat, but not when the whole basal diet replaced alcohol. According to these results, it seems clear that the respiration of hepatic mitochondria is significantly altered when ethanol is supplied to rats at high levels over long periods and when a nutritional basal diet which does not induce fatty liver is used.     

Virulence of Streptococcus mutans: a sensitive method for evaluating cariogenicity in young gnotobiotic rats.

Gnotobiotic rats infected with Streptococcus mutans 6715 at 19 days of age and fed a purified diet (305) containing 5% sucrose developed extensive caries lesions on all molar surfaces within 16 days (35 days of age). Approximately twice as many lesions developed when infected rats were maintained until 45 days of age, whereas noninfected rats did not develop caries when fed diet 305. Gnotobiotic rats infected with S. mutans 6715 and fed a purified diet containing no sucrose (300) until day 25 and subsequently fed diet 305 for 10 days developed lesions similar to rats fed diet 305 for 16 days. Furthermore, rats infected with S. mutans 6715 and fed diet 300 until 45 days of age developed approximately one-half the smooth surface lesions as infected rats fed diet 305 for the same length of time. The level of caries on buccal and proximal molar surfaces in 45-day-old gnotobiotic rats varied when animals were infected with S. mutans AHT, BHT, NCTC 10449, 6715, or LM-7. Animals infected with S. mutans AHT showed more severe lesions on the buccal surfaces than those observed in animals infected with the other strains of S. mutans tested, whereas S. mutans 6715 caused significantly more caries on proximal surfaces. On the other hand, rats infected with S. mutans LM-7 exhibited the lowest level of caries on all molar surfaces of the five strains of S. mutans tested.     

Colonization and cariogenicity of Streptococcus ferus in rats.

Streptococcus ferus, which is indigenous to wild rats, is a member of the mutans group of streptococci. We tested its ability to colonize and to cause caries in laboratory rats by comparing two strains of S. ferus with the very cariogenic Streptococcus sobrinus strain 6715. Groups of rats were fed either finely ground mouse chow or a 56% sucrose diet, or they were switched from chow to the sucrose diet. All three strains colonized the mouths of rats regardless of diet. However, the infectants reached higher proportions of the total flora more quickly in the rats consuming sucrose. Similarly, the percentage of the oral flora represented by an infecting organism increased numerically when rats originally fed chow were switched to the sucrose diet. S. ferus formed plaques on the teeth of the rats, but these plaques did not proliferate over smooth tooth surfaces as extensively as did those of S. sobrinus. Although S. ferus colonized and accumulated, it was non-cariogenic in rats fed sucrose compared both with rats fed similarly but infected with S. sobrinus 6715 and with uninfected controls. In vitro measurements suggested the S. ferus produced acid less rapidly than S. sobrinus. Thus, the lack of cariogenicity in S. ferus may result from an inability to form copious plaques on smooth tooth surfaces and from low acid production and, therefore, may represent a natural absence of the pathogenic potential usually inherent in the mutans streptococci.     

Pectin-enriched diet affects distribution and expression of apoptosis-cascade proteins in colonic crypts of dimethylhydrazine-treated rats

The colonic crypt contains highly proliferative cells in its base and differentiated cells on its luminal surface. Carcinogenesis significantly affects this orderly cellular distribution. The aims of this study were: i) to examine the expression of apoptosis-related proteins along the crypt-lumen axis during 1, 2-dimethylhydrazine (DMH)-induced carcinogenesis, ii) to assess whether a diet supplemented with the soluble fiber pectin affects those parameters, in comparison to non-carcinogen-treated rats and in relation to rats fed a standard diet and treated with DMH. The pectin-enriched diet induced upregulation of active caspase-1 subunit (20 kDa) and of caspase-3 precursor in DMH-treated rats. Pectin enhanced caspase-3 activity in all colonocyte populations, in both non-DMH and DMH-treated rats. The luminal colonocytes exhibited higher caspase-3 activity than proliferative colonocytes of rats fed a standard diet in non-DMH and DMH-treated rats, whereas in pectin-fed non-DMH-treated rats, equal activity was measured among all colonocyte populations. In the DMH-treated rats, the cleaved poly(ADP-ribose) polymerase subunit (89 kDa) was detected in luminal colonocytes of rats fed pectin and was higher than in rats fed the standard diet. Bak was equally expressed in isolated colonocytes from rats of both dietary groups treated with DMH and in the normal rats fed pectin, whereas in the non-DMH-treated rats fed a standard diet, higher expression was obtained in differentiated colonocytes. In the DMH-treated rats, Bcl-2 expression was lower in all colonocytes harvested from rats fed pectin, relative to rats fed the standard diet. Apoptotic index in the DMH-treated groups was higher in rats receiving the pectin diet compared with the standard diet in both the differentiated cell populations and the proliferating colonocytes. Average tumor number and volume per rat were lower in rats fed pectin. These findings indicate that dietary fibers regulate expression, function and distribution of apoptotic-related proteins in the crypt during colon carcinogenesis, changes that probably induce a reduction in tumor volume. We assume that butyrate, produced following fermentation of pectin, may play a key role in these effects.     

Dietary preference behavior in rats fed bitter tasting quinine and sucrose octa acetate adulterated diets

Rats were fed a bitter and presumably toxic quinine adulterated diet and a more bitter, but non-toxic sucrose octa acetate (SOA) adulterated diet. In two-diet preference tests the animals initially preferred the quinine diet over the SOA diet. After being fed only the SOA diet for four days the rats switched their preference to this diet during a subsequent two-diet preference test. In 10 day single diet tests the rats ate significantly more of the SOA diet than of the quinine diet. The results are consistent with the hypothesis that the toxic effects of quinine result in the formation of a conditioned taste aversion to the diet, and indicate that rats can distinguish between two bitter diets and develop a preference for the more bitter, but non-toxic one.     

Random or selective cell death during pancreatic involution following withdrawal of raw soya flour feeding in the rat

Feeding a diet of raw soya flour, which contains a trypsin inhibitor, results in pancreatic hypertrophy and hyperplasia, and when this diet is withdrawn rapid involution ensues. This study examines whether cells produced during the hyperplastic response to raw soya four (RSF) are selectively destroyed during involution following the withdrawal of this diet. Six week old male Wistar rats were fed RSF. At 7, 9 and 12 d after commencing the diet, during the period of maximum cell proliferation, rats were injected with 0.5 muCi/g body weight of tritiated thymidine. The rats were continued on this diet for a further 16d. By 1 mth pancreatic DNA, RNA, protein, weight and the specific activity of pancreatic DNA were all significantly greater in rats fed RSF, compared to control rats fed a standard diet. When rats fed RSF were changed to a diet free of trypsin inhibitor for 7 d, pancreatic DNA, RNA, protein and weight returned to control values, however the specific activity of DNA remained unchanged from the RSF value. This report suggests that cell death involves cells produced before and during the hyperplastic response to RSF, since there is no change in the specific activity of the organ after involution.     

Plasma catecholamines in fasted and sucrose supplemented rats

Rats were food deprived or given a sucrose supplemented diet for 3 days. Resting plasma catecholamine levels measured remotely from undisturbed rats were not altered by either dietary treatment. However, food deprivation did result in decreases in resting mean arterial blood pressure, heart rate, plasma volume and plasma Na+ concentration. After one minute of intermittent footshock food deprived and sucrose fed rats did not differ from controls with respect to blood pressure, heart rate or plasma catecholamine levels but food deprived rats were less active during footshock and had lower levels of plasma glucose immediately after footshock when compared to controls or sucrose fed rats. Food deprivation and dietary sucrose supplementation have been shown to alter norepinephrine (NE) turnover in specific sympathetic target tissues. Our data indicate that these changes in turnover are not reflected by changes in plasma NE. Therefore, NE turnover rates and plasma NE concentration may not be equivalent indices of sympathetic activity.     

Effect of age and long-term diet on exocrine pancreas of the rat.

Effects of age and long-term nutritional treatment on pancreas composition and digestive function were determined in rats fed a cereal-type chow after weaning (experiment 1) or diets with 30% casein and 34% butterfat, 54% starch, or 54% sucrose after 9 mo (experiment 2). The rats were adapted for 1-2 wk to a 15% whole-egg protein diet before killing. In experiment 1, pancreas size, nucleic acid, and digestive enzyme content increased significantly with age up to 3 mo. Values for pancreatic weight and DNA were significantly greater in 28-mo-old rats than in 12-mo-old rats. Pancreatic digestive enzyme content was 65-100% lower in rats with and without gross pathologic lesions. In experiment 2, mortality was higher and pathologic changes were more pronounced by 24 mo in rats fed the butterfat or sucrose diet. Usually, pancreatic enzymes were not reduced as much as in experiment 1, although chymotrypsinogen and trypsinogen concentrations were significantly and negatively correlated with the degree of pathologic change. Apparent digestibility of dietary nitrogen and food energy content was not reduced in rats with reduced enzyme reserves. The rate of incorporation in vitro of label into pancreatic protein and RNA did not differ significantly among aged and control rats.     

Effect of low calcium diet on the ultrastructure of the rat parathyroid gland

Young female rats were fed with normal (1.18%) or low (0.05%) calcium diet for 3, 7, 15 or 30 days. The morphology of the parathyroid glands was studied together with serum calcium, parathyroid hormone (PTH), calcitonin and bone mineral density (BMD). As compared to the animals fed with the normal calcium diet, BMD of whole body of the rats fed with the low calcium diet was significantly decreased, whereas the serum PTH level was increased. The parathyroid glands in the rats fed with the low calcium diet were markedly enlarged. In the parathyroid chief cells of the rats fed with the low calcium diet, the Golgi complexes and the cisternae of the granular endoplasmic reticulum were well developed, while the large granules and large vacuolar bodies decreased. Some secretory granules located near the plasma membrane. A proportionally larger increase of the cytoplasm was estimated in the rats fed with the low calcium diet for three and seven days. Enlargement of the cytoplasm and rather frequent mitoses of the chief cells were observed in the rats fed with the low calcium diet for 15 and 30 days. These findings suggest that the rapid bone loss in young rats induced by the low calcium diet is essentially due to stimulated activity of the parathyroid gland. The stimulated gland may be a result of hypertrophy at the early stage and a combination of hypertrophy and hyperplasia at the later stage of calcium deficiency.     

Effect of a low magnesium diet on in vitro glucose uptake in sucrose fed rats.

Dietary magnesium deficiency and excess sucrose in the diet have been shown to play an important role in the development of insulin resistance. This study is an extension of a previously published experiment (Magnes Res 2004; 17: 293-300) and is focused on the effect of a low magnesium diet on in vitro glucose uptake in sucrose fed rats. For this purpose male Wistar rats were divided into four groups and fed control, high sucrose, low magnesium and high sucrose low magnesium diets for a period of three months. Serum and erythrocyte magnesium values demonstrated a significant drop in the low magnesium and high sucrose low magnesium groups. A significant increase was observed in the body weight of the high sucrose group, whereas the weights of animals in the high sucrose low magnesium group remained unchanged from controls. The biochemical analysis showed a significant decrease in in vitro glucose uptake in liver, muscle and diaphragm of rats consuming high sucrose, low magnesium and high sucrose low magnesium diets. The maximum reduction, however, was observed in the combined high sucrose low magnesium group. These findings seem to suggest the potential of a high sucrose low magnesium diet to cause insulin resistance by reducing glucose uptake in target tissues of rats.     

Effect of soft diet and aging on rat masseter muscle and its motoneuron

To determine the effect of a soft diet and aging on the masticatory motor unit, we investigated the morphologic and metabolic properties of the superficial masseter muscle and its motoneurons in rats. Twenty rats were divided into four groups of five rats: rats fed a hard diet until 4 months after birth (hard, young), rats fed a soft diet until 4 months after birth (soft, young), rats fed a hard diet until 22 months after birth (hard, old), and rats fed a soft diet until 22 months after birth (soft, old). The diameter of the fast-twitch oxidative glycolytic muscle fiber was significantly smaller in the soft than the hard, and in the old than the young groups. The glycolytic enzyme (phosphofructokinase) activity of the muscle was significantly weaker in the old than the young group. There was no significant difference in soma diameter of the motoneurons between the soft and hard group, while the diameter was significantly larger in the old than in the young group. There was no significant difference in NADH-diaphorase activity of the motoneurons between the soft and hard group, while significantly less activity was demonstrated in the old than in the young group. The reduction in motor unit activity caused by the soft diet is considered to influence the morphologic and metabolic properties in the superficial masseter muscle but not in its motoneurons. The reduction in the oxidative enzyme activity of motoneurons with aging may occur regardless of the reduction in motor unit activity. © 1993 Wiley-Liss, Inc.     

Effect of short-term exercise training on muscle glycogen in resting conditions in rats fed a high fat diet

Summary It has been reported that exercise training increases muscle glycogen storage in rats fed a high carbohydrate (CHO) diet in resting conditions. The purpose of this study was to examine whether a 3-week swimming training programme would increase muscle glycogen stores in rats fed a high-fat (FAT) diet in resting conditions. Rats were fed either the FAT or CHO diet for 7 days ad libitum, and then were fed regularly twice a day (between 0800 and 0830 hours and 1800 and 1830 hours) for 32 days. During this period of regular feeding, half of the rats in both dietary groups had swimming training for 3 weeks and the other half were sedentary. The rats were not exercised for 48 h before sacrifice. All rats were killed 2 h after their final meal (2030 hours). The glycogen contents in red gastrocnemius muscle, heart and liver were significantly higher in sedentary rats fed the CHO diet than in those fed the FAT diet. Exercise training clearly increased glycogen content in soleus, red gastrocnemius and heart muscle in rats fed the CHO diet. In rats fed the FAT diet, however, training did not increase glycogen content in these muscles or the heart. Exercise training resulted in an 87% increase of total glycogen synthase activity in the gastrocnemius muscle of rats fed the CHO diet. However, this was not observed in rats fed the FAT diet. The total glycogen phosphorylase activity in the gastrocnemius muscle of the rats of both dietary groups was increased approximately twofold by training. These results suggested that muscle glycogen was enhanced in rats fed the CHO diet and that the glycogen content of the muscle of rats fed the FAT diet was not increased by exercise training.