Hepatic iron accumulation in copper-deficient rats

Studies of anaemia and tissue iron distribution were carried out in copper-deficient rats and pair-fed control animals given Fe orally or parenterally in varying doses. The anaemia of Cu deficiency was partially but incompletely corrected by oral Fe supplementation of one- to five-fold normal dietary levels or by intramuscular Fe supplementation. Serum Fe increased in Cu-deficient animals as the dose of supplemental Fe was increased. Hepatic Fe accumulation occurred in Cu-deficient rats which were administered with either oral Fe in two- to five-fold excess or low doses of intramuscular Fe. This difference was not seen in animals receiving high doses of intramuscular Fe, but similar relative differences were seen in Cu-deficient and Cu-replete rats which had been given no Fe supplementation. Duodenal Fe was not increased in Cu deficiency. Bone marrow Fe was present in Cu-deficient animals receiving either parenteral or oral Fe supplementation. Present studies suggest that a decrease in caeruloplasmin (EC 1.16.3.1) activity does not wholly explain the anaemia of Cu deficiency. Fe accumulation may be restricted to the liver, suggesting that Cu may be required for normal intracellular Fe metabolism.     

Development of anemia in copper-deficient rats fed high levels of dietary iron and sucrose

The effects of dietary carbohydrate and iron on the development of copper deficiency were examined. Male Sprague-Dawley rats (n = 48) were limit-fed one of eight diets in a 2 X 2 X 2 factorial design for 19 d. Two levels of copper (0.85 or 8.6 micrograms Cu/g diet) and iron (54 or 226 micrograms Fe/g diet) and two types of carbohydrate (sucrose or cornstarch, 65.3%) were fed. Compared with control rats, copper-deficient rats had lower hematocrits, lower ceruloplasmin levels, lower tissue levels of copper and increased hepatic iron levels. Copper-deficient rats fed sucrose had significantly lower hematocrits, lower apparent absorption of copper, lower liver iron levels and higher plasma triglyceride levels than copper-deficient rats fed cornstarch. Copper-deficient rats fed sucrose with 226 micrograms Fe/g diet had hematocrit levels that were 15% lower than all other copper-deficient levels and 23% lower than control levels. Tissue levels of copper among copper-deficient rats were not affected by the type of carbohydrate or by the level of dietary iron. These data indicate that both high iron and sucrose can affect the development of the copper deficiency.     

Impairment of glucose tolerance in copper-deficient rats: dependency on the type of dietary carbohydrate

Copper deficiency was induced in weanling rats in order to study the possible interaction between the types of dietary carbohydrate and copper deficiency on glucose tolerance. Weanling male rats were fed copper-deficient or copper-supplemented diets containing either 62% starch, fructose or glucose. During week 5 the fructose portion of the copper-deficient diet was replaced (20 rats) by starch (10 rats) or glucose (10 rats). During the 9th week, an oral glucose tolerance test weas performed. Copper deficiency was associated with impaired glucose tolerance characterized by increased blood glucose and decreased insulin levels only in copper-deficient rats fed the monosaccharides fructose or glucose but not the polysaccharide starch. Changing the dietary carbohydrates in the copper-deficient diet from fructose to starch increased insulin levels and decreased blood glucose in response to the glycemic stress when compared to rats continuously fed fructose. Although both glucose and fructose feeding impaired the glucose tolerance, fructose was more diabetogenic. This could be demonstrated by some improvements in glucose tolerance when the copper-deficient rats were switched from the fructose to the glucose diet. The data indicate that copper deficiency per se does not impair glucose tolerance.     

Adverse effects of high dietary iron and ascorbic acid on copper status in copper-deficient and copper-adequate rats

The effects of elevated dietary ascorbic acid and iron on copper utilization were examined. Male Sprague-Dawley rats were fed one of two levels of Cu (deficient, 0.42 microgram Cu/g, or adequate, 5.74 micrograms Cu/g), Fe (moderate, 38 micrograms Fe/g or high, 191 micrograms Fe/g), and ascorbic acid (low, 0% or high, 1% of the diet) for 20 d. High Fe decreased (p less than 0.05) Cu absorption only in Cu-deficient rats. High ascorbic acid significantly decreased tissue Cu levels in Cu-adequate rats. High Fe with ascorbic acid caused severe anemia in Cu-deficient rats and decreased plasma ceruloplasmin by 44% in Cu-adequate rats. Cu,Zn-superoxide dismutase activity in erythrocytes was decreased (p less than 0.05) by 14% during Cu deficiency but was not affected by Fe or ascorbic acid. These results may be important to individuals with high intakes of Fe and ascorbic acid.     

Redistribution of abdominal fat after a period of food restriction in rats is related to the type of dietary fat

The aim of the present experiment was to test the hypothesis that during refeeding a redistribution of intra-abdominal fat takes place and that both the recovery of weight and the redistribution of intra-abdominal fat are related to the type of dietary fat. The experimental study was carried out using male Sprague-Dawley rats. Three groups of animals were fed diets with three different fatty acid profiles. Each group contained two branches, one fed normally and the other fed initially with a 50 % energy reduction followed by refeeding ad libitum with the same isoenergetic diet as the control branch, giving a total of six treatments. Measurements were made of the final and incremental weight of the rat, weight of the intra-abdominal adipose tissue (total intra-abdominal, epididymal, omental and retroperitoneal adipose tissue weight), and feed efficacy (weight increment/metabolizable energy intake). Carcass, epididymal, omental, and muscle lipid contents, carcass protein and energy density were also measured. The results revealed that diets rich in fish oil or olive oil increase catch-up growth more than diets rich in saturated fats. During refeeding the lipid content in the adipose tissue increases while that of muscle tissue decreases. A diet rich in saturated fats induces a relative increase in the amount of intra-abdominal adipose tissue. The lipid content in adipose and muscle tissues and the distribution of intra-abdominal fat can all be modified by the type of dietary fat.     

Plasma esterase-1 (ES-1) activity in rats is influenced by the amount and type of dietary fat, and butyryl cholinesterase activity by the type of dietary fat.

In previous work, we studied, under conditions of ad libitum food consumption, the effect of amount and type of dietary fat on plasma esterase-1 (ES-1) and butyryl cholinesterase activity in rats. This was done by the isoenergetic replacement of dietary fat by carbohydrates or by another fat source. The observed change in enzyme activity could theoretically be determined by either the dietary omission or the addition or by the combination. In the present work, we studied under restricted feeding conditions the effect of supplemental energy in various forms to determine the effect of the supplement alone. Supplemental coconut fat, but not isoenergetic amounts of either glucose or casein, raised plasma ES-1 activity. None of these supplements influenced butyryl cholinesterase activity. In a second experiment, we demonstrated that the ES-1 enhancing effect of supplemental coconut fat also occurred with fish oil, whereas the stimulatory effects of olive oil and corn oil were less pronounced. Supplemental fish oil, but not the three other fats, significantly reduced the depression in butyryl cholinesterase activity. Plasma cholesterol concentration was negatively associated with butyryl cholinesterase activity, but was not related to ES-1 activity. The two esterases were not correlated with plasma triglyceride concentration. We conclude that both the amount and type of fat in the diet of rats have specific influences on plasma ES-1 activity and that butyryl cholinesterase activity is affected by the type of fat.     

Role of dietary fat type in the development of adiposity from dietary obesity-susceptible Sprague-Dawley rats

The present study was designed to define how dietary fat type regulates body adiposity in dietary obesity-susceptible (DOS) Sprague-Dawley (SD) rats. Eighty-three SD rats received a purified diet containing 50 g maize oil (MO)/kg for 3 weeks and then thirty-nine of the rats, designated as the DOS rats, were allotted to diets containing 160 g MO (DOS-MO), beef tallow (DOS-BT) or fish oil (DOS-FO)/kg for 9 weeks. As a result of the experiment, the DOS-FO rats had significantly (P<0.05) reduced weight gain and abdominal and epididymal fat-pad mass than the DOS-MO and DOS-BT rats. Serum leptin level was also significantly (P<0.05) lower in the DOS-FO rats; however, hypothalamic leptin receptor (a and b) mRNA and neuropeptide Y expressions were not altered by dietary fat sources. A lower acetyl-CoA carboxylase mRNA expression in the liver was observed in the DOS-FO group, whereas hepatic peroxisome proliferator-activated receptor-gamma mRNA and protein expressions were markedly elevated in the DOS-FO group compared with those in the other groups. We did not observe differences in acetyl-CoA carboxylase and peroxisome proliferator-activated receptor-gamma expressions in epididymal fat of the DOS rats consuming MO, BT or FO. It is concluded from our present observations that dietary fat type, especially that rich in FO, plays a potential role in down-regulation of adiposity by altering hepatic lipogenic genes, rather than feeding behaviour, in the DOS-SD rats.     

The lowering effect of dietary glucose versus starch on fat digestibility in rats is dependent on the type of fat in the diet

The aim of the study was to determine whether the type of dietary fat influences the effect of dietary glucose on lipid digestibility. Earlier work had shown that glucose, when compared with starch, reduced fat digestibility in rats fed rations containing animal fat as fat source. Male rats (n = 6/group) were fed for two weeks on purified diets containing either 62% (w/w) starch or glucose and either 8% (w/w) palm oil, coconut fat, soybean oil or medium-chain triglycerides (MCT) as the main source of fat. The diets had no differential effect on growth. Glucose significantly depressed apparent lipid digestibility in rats fed the diets containing either palm oil or coconut fat, but not in rats given the diets containing either soybean oil or MCT. Thus, the inhibitory effect of glucose on lipid digestibility in rats is dependent on the dietary lipid source. This observation may contribute to understanding the mechanism by which dietary glucose inhibits fat digestion.     

Effect of dietary cholesterol in normolipidemic subjects is not modified by nature and amount of dietary fat

A double-blind, cross-over study of the effects of adding cholesterol to the diet, equal to about two egg yolks daily, was carried out in 25 men with an average plasma cholesterol of 5.3 mmol/L. There were no significant elevations in the mean plasma concentrations of cholesterol, low-density-lipoprotein cholesterol, or apolipoprotein B. The absence of an effect was seen against two background diets, one high in saturated fatty acids and the other modified in fat content and in fatty acid composition. Although the mean high-density-lipoprotein (HDL) cholesterol concentration did not change, the proportion of HDL2 particles (radius greater than 4.4 nm) increased significantly, especially in subjects who did not show a rise in plasma cholesterol; this may represent a means of clearing dietary cholesterol. The results suggest that reducing dietary cholesterol might be emphasized for hypercholesterolemic subjects rather than for the population generally.     

Iron injection restores brain iron and hemoglobin deficits in perinatal copper-deficient rats

Copper (Cu) deficiency during perinatal development in rats is associated with anemia, lower plasma iron (Fe), and brain Fe. Experiments were conducted to inject Fe dextran into Cu-deficient (Cu-) rat pups to attempt to reverse these conditions. Previous work with older Cu- rats did not reverse anemia following Fe injection. Dams began Cu-adequate (Cu+) or Cu- dietary treatments starting at embryonic d 7 and lasting through weaning. In Expt. 1, pups from each dietary treatment were given a single dose of Fe, 20 mg Fe/kg, or saline (S) at postnatal d 11 (P11). Plasma Fe and hemoglobin were higher in the Fe-injected groups at P13. Brain Fe deficit and brain transferrin receptor enhancement were eliminated in the Cu- group injected with Fe compared with Cu-S pups, supporting an association between low plasma Fe and low brain Fe. In Expt. 2, Fe treatment was increased to 45 mg Fe/kg. Four injections were given between P5 and P18 (total dose, 5-7 mg Fe). At P20, Fe concentrations in 4 brain regions (cortex, cerebellum, medulla/pons, and hypothalamus) generally were higher in all groups than in Cu-S pups. At P25, impaired vibrissae-elicited foot placement was evident in Cu-S rats and was not improved by Fe injection. However, at P26, the brain Fe deficit in Cu-S pups was eliminated by Fe injection. Fe injections in Cu- pups raised plasma Fe, brain Fe, and hemoglobin but did not reverse low cytochrome c oxidase or abnormal striatal behavior.     

Plasma C-reactive protein concentration is not affected by isocaloric dietary fat reduction

OBJECTIVE: There is a strong correlation between plasma C-reactive protein (CRP) concentration and risk of cardiovascular death. Low-fat diets have been recommended for maintenance of cardiovascular health, and it is known that a low-fat diet associated with weight loss lowers CRP concentration. However, it remains unclear whether dietary fat has an effect independent from weight change on markers of inflammation. METHODS: Sixteen overweight subjects who were 46 +/- 14 y old were placed on a weight-maintaining baseline diet consisting of 35% fat, 45% carbohydrate, and 20% energy as protein. After 2 wk, subjects were switched to an isocaloric low-fat diet consisting of 15% fat, 65% carbohydrate, and 20% protein for another 2 wk. For the final 12 wk of the study, subjects consumed the same 15% fat diet ad libitum. At the end of each diet phase, CRP was measured by a high-sensitivity CRP assay. RESULTS: The weight of subjects remained stable during the first 4 wk of isocaloric diets. Plasma CRP concentrations after 2 wk on the weight-maintaining 35% fat diet and 2 wk on the isocaloric 15% fat diet were not significantly different (median +/- interquartile range 1.42 +/- 3.30 and 1.59 +/- 3.29 mg/L, respectively). Three months of ad libitum low-fat diet consumption resulted in a 4.1 +/- 0.7 kg weight loss associated with a decrease in CRP concentration to 1.17 +/- 2.03 mg/L (P = 0.03). CONCLUSION: Loss of body weight decreases CRP concentration, but a decrease in dietary fat without a concurrent change in body weight does not affect CRP concentration in overweight healthy subjects.     

Insulin action in rats is influenced by amount and composition of dietary fat

The chronic influence of dietary fat composition on obesity and insulin action is not well understood. We examined the effect of amount (20% vs 60% of total calories) and type (saturated vs polyunsaturated) of fat on insulin action and body composition in mature male rats. Six months of feeding a high fat (HF) diet led to obesity and impaired insulin action (determined by a euglycemic-hyperinsulinemic clamp), neither of which were reversed by a subsequent 6 months of feeding a low fat (LF) diet. Within HF fed rats, type of fat did not affect body composition or insulin action. Six months of feeding a low fat diet led to only a slight decline in insulin action, with no difference due to type of dietary fat. From 6-9 months, insulin action became more impaired in LF rats fed the saturated diet than in LF rats fed the polyunsaturated diet. By 12 months, all groups were obese and had a similar impairment in insulin action. The amount and type of fat in the diet did not influence the overall degree of impairment in insulin action but did affect the time course. Both feeding a high fat diet and feeding a low fat saturated diet accelerated the impairment in insulin action relative to rats fed a low fat polyunsaturated fat diet.     

Serum cholesterol levels are not elevated in young copper-deficient rats, mice or brindled mice

Experiments were conducted with suckling male C57BL mice and Sprague-Dawley rats to investigate the relationship between copper deficiency and elevated serum cholesterol. Brindled mice, which have a genetic defect that affects copper distribution, were compared to their normal brothers. Dietary copper deficiency was produced in dams heterozygous for the brindled gene, in normal mouse dams and in rat dams. The subsequent male offspring were compared to those from copper-supplemented dams. Copper deficiency, as assessed by liver copper levels or ceruloplasmin activity, was demonstrated in 12-d-old rats, brindled mice, and in genotypically normal mice from dams fed the copper-deficient diet. However, serum cholesterol levels were not elevated in these "copper-deficient" rats or mice. In one experiment serum cholesterol levels of brindled mice were significantly lower than that of their littermate controls. An additional study was done with older mice. Their dams were fed a low copper diet from parturition throughout lactation, and the pups were fed the same copper-deficient diet for 4 wk after weaning. The 7-wk-old male copper-deficient mice had liver copper levels below 1 microgram/g, but no elevation in serum cholesterol was observed. The failure to demonstrate a rise in serum cholesterol in these perinatal models may be due in part to less severe hepatic copper deficiency because of neonatal copper reserves in liver.     

Excess dietary L-cysteine, but not L-cystine, is lethal for chicks but not for rats or pigs

A comparative species investigation of the relative pharmacologic effects of sulfur amino acids was conducted using young chicks, rats, and pigs. Ingestion of excess Met, Cys, or Cys-Cys supplemented at 2.5-, 5.0-, 7.5-, or 10 times the dietary requirement in a corn-soybean meal diet depressed chick growth to varying degrees. Strikingly, ingestion of excess Cys at 30 g/kg Cys (7.5-times the dietary requirement) caused a chick mortality rate of 50% after only 5 d of feeding. Growth was restored and chick mortality was reduced by supplementing diets containing 25 g/kg excess Cys with KHCO3 at 10 g/kg. Additionally, mortality was prevented by supplementing the drinking water of chicks receiving 25 g/kg supplemental Cys with H2O2 (0.05% final concentration). After young rats and pigs consumed excess Cys or Cys-Cys up to 40 g/kg for 14 d, weight gain was severely depressed, but we observed no mortality. An excess of dietary Cys-Cys>or=48 g/kg caused some mortality in rats. Pigs exhibited rapid recovery from growth-depressing excesses of Cys or Cys-Cys. These results lend credence to the acute toxic effects associated with the ingestion of excess sulfur amino acids and highlight the potential for excess dietary cyst(e)ine to be more pernicious than Met in certain species.     

Interrelated effects of the type of dietary fat and carbohydrate on cholesterol metabolism in rats

Rats were fed semipurified diets, differing in the amount of cholesterol and the sources of fat (corn oil or coconut fat) and carbohydrate (sucrose or starch). After 21 days dietary corn oil had induced higher serum cholesterol concentrations than did coconut fat, except for on diets with a high-cholesterol, high-sucrose background. On high-cholesterol diets containing coconut fat, dietary sucrose increased serum cholesterol, when compared with starch; with corn oil sucrose tended to lower serum cholesterol. Such a tendency was not seen on cholesterol-free diets. Cholesterol feeding caused a dramatic increase in liver cholesterol with all dietary carbohydrate-fat combinations. Liver cholesterol was higher in rats fed corn oil than in those fed coconut fat. The influence of the type of dietary carbohydrate on liver cholesterol was dependent on the type of fat and the amount of cholesterol in the diet. Thus effects of a single dietary component on serum and liver cholesterol in rats are strongly influenced by the background of the diet. This implies that published results of experiments on diet and cholesterol metabolism in rats cannot be compared readily.     

Dietary fat level affects tissue iron levels but not the iron regulatory gene HAMP in rats

Because dietary fats affect the regulation and use of body iron, we hypothesized that iron regulatory and transport genes may be affected by dietary fat. A model of early-stage I to II, nonalcoholic fatty liver was used in which rats were fed standard (35% energy from fat) or high-fat (71% energy from fat) liquid diets with normal iron content (STD/HF groups). In addition, intraperitoneal injections of iron dextran were given to iron-loaded (STD+/HF+ groups) and iron-deficient diets to STD−/HF− groups. Plasma osmolality, hemoglobin level, and mean corpuscular hemoglobin concentration were increased in all STD diet groups compared with all HF diet groups. Plasma iron and transferrin saturation were affected by an interaction between dietary fat and iron. They were high in the STD group (normal iron) compared with their respective HF group. Similarly, this group also showed a 4-fold increase in the messenger RNA expression of the hepatic hemochromatosis gene. Spleen iron was high in the iron-loaded STD+ group compared with all other groups. Hepatic iron and messenger RNA expression of peroxisome proliferator–activated receptor-γ, CCAAT/enhancer binding protein α, interleukin-6, and iron transport genes (transferrin receptor 2, divalent metal transporter 1 iron-responsive element, and divalent metal transporter 1 non–iron-responsive element) were increased, whereas tumor necrosis factor α was decreased in the HF diet groups. The expression of iron regulatory gene HAMP was not increased in the HF diet groups. Iron regulatory and transport genes involved in cellular and systemic iron homeostasis may be affected by the macronutrient composition of the diet.