Resistance Exercise Training Improves Metabolic and Inflammatory Control in Adipose and Muscle Tissues in Mice Fed a High-Fat Diet

This study investigates whether ladder climbing (LC), as a model of resistance exercise, can reverse whole-body and skeletal muscle deleterious metabolic and inflammatory effects of high-fat (HF) diet-induced obesity in mice. To accomplish this, Swiss mice were fed for 17 weeks either standard chow (SC) or an HF diet and then randomly assigned to remain sedentary or to undergo 8 weeks of LC training with progressive increases in resistance weight. Prior to beginning the exercise intervention, HF-fed animals displayed a 47% increase in body weight (BW) and impaired ability to clear blood glucose during an insulin tolerance test (ITT) when compared to SC animals. However, 8 weeks of LC significantly reduced BW, adipocyte size, as well as glycemia under fasting and during the ITT in HF-fed rats. LC also increased the phosphorylation of Akt_Ser473 and AMPK_Thr172 and reduced tumor necrosis factor-alpha (TNF-α) and interleukin 1 beta (IL1-β) contents in the quadriceps muscles of HF-fed mice. Additionally, LC reduced the gene expression of inflammatory markers and attenuated HF-diet-induced NADPH oxidase subunit gp91phox in skeletal muscles. LC training was effective in reducing adiposity and the content of inflammatory mediators in skeletal muscle and improved whole-body glycemic control in mice fed an HF diet.     

Effects of diet and ketones on rat pancreatic lipase in cultured acinar cells

The activity, synthesis rate and mRNA level of pancreatic lipase increase with dietary fat intake. Ketones, intermediates of lipid metabolism, have been proposed to mediate this change. Therefore, we investigated their direct effect on cultured pancreatic acinar cells and examined their possible interactive effects with glucose and dietary fat. beta-Hydroxybutyrate (0.01 to 2 mmol/L) did not affect lipase activity in cells isolated from rats fed a commercial nonpurified (NP) diet and cultured in high glucose (HG, 27.8 mmol/L) or low glucose (LG, 6.9 mmol/L) medium. The effects of ketones were also examined in acinar cells isolated from rats fed purified high fat (HF, 67% of energy from fat) or low fat (LF, 11% of energy from fat) diet. Cellular lipase was significantly higher in cells from HF-fed rats at both 24 and 48 h (264% and 145% of LF values, respectively; P less than 0.0001). beta-Hydroxybutyrate significantly increased (P less than 0.04) lipase activity in LF cells at 48 h but did not affect lipase activity in HF cells. These studies suggest that ketones may be involved in the regulation of pancreatic lipase in rats fed a LF diet, but their role is complex and interactive with dietary carbohydrate and fat.     

A controlled high-fat diet induces an obese syndrome in rats

The prevalence of obesity is increasing. Although the etiology of obesity is complex, dietary factors, particularly the consumption of a high-fat (HF) diet, is considered a risk factor for its development. Nonetheless, a causal role of dietary fat has never been definitively documented, in part because of inadequate animal models. We developed a rat model of diet-induced obesity that will be a powerful tool for assessment of this issue. In four experiments, Long-Evans rats ate ad libitum a synthetic semipurified diet containing 20 g (HF) or 4 g [low-fat (LF)] of fat/100 g of diet or a nonpurified diet. Other rats ate ad libitum the HF diet in amounts matched to the energy intake of the LF rats. When compared over 10 wk of free feeding, HF rats weighed 10% more (P < 0.01) than LF rats and had 50% more body fat (P < 0.01), as well as significant hyperleptinemia and insulin resistance. Compared with rats fed the nonpurified diet, the HF rats had even more marked differences in these variables. The rats fed the HF diet to match the rats fed the LF diet had similar body weights but significantly more adipose tissue than LF rats, suggesting that diet composition and/or energy density of the diet affects fat deposition. This dietary regimen has reproducible effects on body size and composition, and these are similar in male and female rats. This model of diet-induced obesity will be a useful tool for studying the mechanisms by which dietary fat influences the regulation of energy balance.     

Plasma and muscle carnitine in experimental uremia

The effect of uremia and dietary protein intake on carnitine levels in skeletal muscle and plasma of rats was evaluated. Male Sprague-Dawley rats weighing 160–180 g were made chronically uremic by partial ligation of left renal artery and contralateral nephrectomy. Rats were fed diets containing 8% or 18% casein ad libitum for 8 weeks and sham-operated pair-fed animals were used as controls. At the end of the experiment, all animals were fasted overnight and decapitated. Skeletal muscle and plasma were assayed for free, total acid-soluble and long-chain acylcarnitines. The concentration of total carnitines in skeletal muscle were similar in uremic and control rats fed either 8% or 18% casein diet. However, the muscle long chain acylcarnitines were higher in rats fed the 18% casein diet than those fed the 8% casein diet. Diets (8% or 18% casein) had no effect on the plasma carnitine level, although uremic rats had lower levels than the controls. This study has shown that kidney dysfunction may affect the plasma carnitines level but not the total skeletal muscle carnitines concentration.     

Macronutrient selection in an animal model of cholestatic liver disease

Diet selection was investigated in an animal model of cholestatic liver disease produced by bile duct ligation. Animals self-selected diets from separate sources of macronutrients (protein, fat, carbohydrate). Diet selection was evaluated when the fat source was comprised of either a primarily medium-chain fat (coconut oil) or a primarily long-chain fat (Crisco vegetable shortening). Relative to intakes of control animals, bile duct ligated (BDL) animals consuming the long-chain fat decreased fat intake, decreased protein intake, and increased carbohydrate intake. Consumption of the fat source was decreased in BDL rats fed the medium-chain fat relative to intakes of control animals, however carbohydrate and protein intakes were not affected. Total caloric intake was comparable to control intakes by day 16 post-ligation in BDL rats fed the long-chain fat and by day 11 in BDL rats fed the medium-chain fat. Body weight gain was significantly greater in BDL rats fed the medium-chain fat than in those fed the long-chain fat. Mortality was 44% in BDL animals fed the long-chain fat, and 0% in those fed the medium-chain fat. The results suggest that BDL animals make dietary selections which may decrease the severity of liver disease. Differences between ligated animals consuming either medium- or long-chain fats suggest that some fat sources may be more beneficial during cholestasis.     

Effect of feeding grape pomace on selected metabolic parameters associated with high fructose feeding in growing Sprague-Dawley rats

The effect of feeding grape pomace on certain metabolic parameters associated with high fructose (HF) feeding was studied. Forty male growing Sprague-Dawley rats were randomly assigned into groups: (1) control; (2) HF; (3) HF with low-level (1.5% of diet) grape pomace (HF+LP), and (4) HF with high-level (5.0% of diet) grape pomace (HF+HP). The HF+LP and HF+HP diets provided 115 and 218 mg of procyanidins/kg, respectively. Compared with the controls, HF-fed animals consumed less and were smaller, whereas animals in the HF+LP and HF+HP groups were in between. A similar trend was observed for abdominal fat and abdominal fat as a percentage of body weight. No change in heart or kidney weight occurred. Liver weight as a percentage of body weight was higher for animals when fructose was included in the diet compared with those on control diet, and inclusion of grape pomace had no effect. Fasting plasma glucose, insulin, and triglyceride levels tended to be higher in animals fed HF diet, and grape pomace reduced their levels to values similar to the control animals. Compared with control animals, HF-fed animals had higher weekly postprandial plasma triglycerides, which were reduced by feeding grape pomace, but no change in plasma cholesterol was observed. Glucose intolerance was observed in animals fed HF diet and was accompanied by a 25% increase in homeostatic model assessment (HOMA) of insulin resistance. Inclusion of grape pomace increased glucose tolerance and insulin sensitivity. No significant change (P>.1) in HOMA of β-cell function or Quantitative Insulin-Sensitivity Check Index was observed. Overall, HF diet did not produce as strong a response of metabolic syndrome as has been shown in the literature. The inclusion of grape pomace in the diet was effective in modulating some aspects of metabolic parameters associated with metabolic syndrome, and the higher level of grape pomace in the diet produced a slightly better response than the lower level.     

Oxidative stress and antioxidant status in rat blood, liver and muscle: effect of dietary lipid, carnitine and exercise

The purpose of this study was to determine the effect of dietary fat, carnitine supplementation, and exercise on oxidative damage and antioxidant status. Male Wistar rats (60 days old) were fed diets containing either hydrogenated fat (HF) or peanut oil (PO) with or without 0.5 mg % (of dry diet) carnitine. The rats were given exercise, i.e. swimming for 60 minutes, for 6 days/week for 6 months under each dietary condition. The blood malondialdehyde (MDA) level was higher in PO-fed rats, more so in exercising ones, while the same was not altered in carnitine-supplemented rats irrespective of the dietary fat or physical activity. The MDA level was significantly decreased in muscle, while increased in liver, of carnitine-fed rats. The blood glutathione (GSH) level also significantly increased in exercising rats as compared to sedentary ones, while carnitine supplementation elevated it in all the groups. Exercise and carnitine supplementation significantly lowered GSH levels in liver while increasing it in muscle. The glutathione peroxidase (GPX) activity was significantly increased in blood and muscle from PO-fed exercising rats as compared to sedentary ones, while carnitine supplementation elevated GPX activity in all the groups. The liver and muscle catalase (CAT) activities were significantly increased in PO-fed exercising rats, while carnitine did not have any effect. The pro-oxidative effect of the monounsaturated fatty acid (MUFA)-rich PO diet and prolonged regular exercise was less pronounced due to augmented antioxidant enzymes, GPX and CAT, induced by training to protect against the oxidative stress, while carnitine supplementation could help to counter lipid peroxidation due to exercise through redistribution of GSH from liver to blood and muscle.     

Bitter melon (Momordica charantia) reduces adiposity,lowers serum insulin and normalizes glucose tolerance in rats fed a high fat diet

Bitter melon (BM) is known for its hypoglycemic effect but its effect on rats fed a hyperinsulinemic high fat diet has not been examined. In a dose-response (0.375, 0.75 and 1.5%) study, oral glucose tolerance was improved in rats fed a high fat (HF; 30%) diet supplemented with freeze-dried BM juice at a dose of 0.75% or higher (P < 0.05). At the highest dose, BM-supplemented rats had lower energy efficiency (P < 0.05) and tended (P = 0.10) to have less visceral fat mass. In a subsequent experiment, rats habitually fed a HF diet either continued to consume the diet or were switched to a HF+BM, low fat (LF; 7%) or LF+BM diet for 7 wk. BM was added at 0.75%. Final body weight and visceral fat mass of the two last-mentioned groups were similar to those of rats fed a LF diet for the entire duration. Rats switched to the HF+BM diet gained less weight and had less visceral fat than those fed the HF diet (P < 0.05). The addition of BM did not change apparent fat absorption. BM supplementation to the HF diet improved insulin resistance, lowered serum insulin and leptin but raised serum free fatty acid concentration (P < 0.05). This study reveals for the first time that BM reduces adiposity in rats fed a HF diet. BM appears to have multiple influences on glucose and lipid metabolism that strongly counteract the untoward effects of a high fat diet.     

The effect of 'fibre-filler' (F-plan diet) on iron, zinc and calcium absorption in rats

1. The availability of iron, zinc and calcium in a diet containing 400 g 'fibre-filler' (a mixture of bran, fruit and nuts, used in the F-plan diet)/kg diet (HF diet) was measured by whole-body counting in rats, using 59Fe, 65Zn and 47Ca as extrinsic labels, and compared with a diet of similar mineral content but no 'fibre-filler' (LF diet). Absorption of Fe and Ca was significantly higher from the HF than from the LF diet but there was no difference in Zn availability between the two diets. 2. The ability of rats given LF or HF diets for 3 or 28 d to absorb Fe, Zn and Ca was measured using ferrous sulphate, zinc chloride and calcium chloride in a cooked starch-sucrose (1:1 w/w) paste, extrinsically-labelled with the appropriate isotope. There was no difference in Fe absorption between the HF- and LF-fed groups but both Zn and Ca absorption were higher in LF- than in HF-fed animals after 3 and 28 d. 3. The mineral status of the animals given HF or LF diets for 28 d was examined, and there were no differences in blood haemoglobin, liver and bone Zn and plasma and bone Ca levels. The total liver Fe was significantly lower (P less than 0.05) in the HF-fed animals. 4. It was concluded that 'fibre-filler' did not have an adverse effect on Fe, Zn or Ca metabolism in rats although the long-term effect on Fe status warrants more detailed investigation. Further work is required to extend these studies to man.     

Body fat deposition: effects of dietary fat and two exercise protocols.

We evaluated the effects of two levels of dietary fat (0 and 20g beef tallow/100g diet) and two treadmill exercise protocols (low-intensity, high-intensity) on fat deposition in rats.

Male Wistar rats (n = 50) remained sedentary or were forced to run 840 meters/day, 5 days/week, on a rodent treadmill. Those on the high-intensity protocol covered this distance in less time (38 min) than those on the low-intensity program (60 min). Responses to high-fat (HF) and low-fat (LF) diets were compared within each exercise group for this 8-week study.

High fat feeding, as a single factor, did not affect energy intake, carcass fat, intramuscular fat, or fat associated with any tissue studied. The HF diet also did not affect responses to either exercise protocol. The high-intensity-exercised animals had less carcass fat (LF: 21% less; HF: 33% less), smaller omental fat pads (LF: 20% less; HF: 37% less), and retroperitoneal fat pads (LF: 19% less; HF: 38% less), and lower serum triglyceride levels (LF: 26% less; HF: 41% less) than sedentary rats. Those differences were less marked for the low-intensity-exercise rat. Neither mode of exercise or diet affected lipid concentrations in hindlimb muscles, livers, hearts, or kidneys.

Exercised animals generally had less fat deposition than sedentary rats but this was more pronounced for high-intensity than low-intensity-exercised rats, and was not affected by feeding a 20% beef tallow diet.     

Dietary cholesterol and experimental mammary cancer development

The effect of high‐ and low‐fat diets, with and without cholesterol supplementation, on the development of N‐methylnitrosourea (NMU)‐induced mammary tumors was assessed. Diets consisting of 1. high fat (HF) (20% lard), 2. HF + cholesterol, 3. low fat (LF) (4% lard) + cholesterol, and 4. LF were fed to F344 female rats (24 animals/group) 2 days after NMU administration, and cumulative mammary tumor incidence was monitored for a total of 26 weeks. Animals fed HF diets exhibited significantly greater tumor incidences and numbers of tumors/animal than did animals fed LF diets (p < .0001), regardless of whether cholesterol was present in the diet. These results are consistent with the hypothesis that the mammary‐tumor promoting effects of HF diets are exerted primarily by the triglyceride fraction rather than by the nonsaponifiable (sterol) fraction of total dietary fat.     

Lipoprotein lipid and protein responses to dietary fat and diabetes in rats

Dietary and insulin-deficiency types of hyperlipidemia were compared in adult normal and streptozotocin-induced diabetic male breeder rats. High beef tallow, high corn oil or low fat diets (BT, CO and LF, respectively) were fed ad libitum for 2 months. Glucose and insulin were measured in plasma and total cholesterol, free cholesterol, cholesteryl ester, triglycerides and apoproteins in very low density, low density and high density lipoproteins (VLDL, LDL and HDL, respectively). Diet did not affect plasma glucose or insulin levels. LDL-triglycerides were higher in BT and diabetic than in CO and LF rats. HDL-free cholesterol levels were higher in CO- and LF-than in BT-fed rats. Diabetes resulted in a decrease in HDL-cholesterol. Diabetic animals had higher HDL-apoA-I (apolipoprotein A-I) levels than did CO- and LF- but not BT-fed rats. VLDL-triglycerides were higher in diabetic than in normal rats, with no dietary differences in normal rats. In LDL, apoB levels were lower and apoE levels were higher in LF-fed rats than in animals fed high fat diets. Diabetes resulted in an increase in LDL-apoB but a decrease in LDL-apoE. HDL-apoE levels were higher, although HDL-apoA-I levels were lower in LF than in high fat-fed rats. The results related to lipoprotein composition supported the hypothesis that excess intake of a diet high in saturated fat may contribute to a metabolic pattern that resembles that of a diabetic state.     

Exercise-trained but not untrained rats maintain free carnitine reserves during acute exercise

Exercise training is known to induce physiological adaptations that improve exercise performance and alter patterns of energy substrate utilization to favour fatty acid oxidation. L-Carnitine is an essential cofactor for the oxidation of fatty acids under all physiological conditions, including exercise. This study evaluated the effect of acute exercise on carnitine concentrations in tissue and serum, liver carnitine palmitoyltransferase-I activity and expression, and serum lipids in both trained and untrained rats as compared to non-exercised rats. Serum acyl- and total carnitine was significantly higher in the trained animals, whether exercised or not, suggesting an exercise-induced increase in a renal threshold for carnitine. Untrained rats had significantly higher acylcarnitine in skeletal muscle and an acyl/free carnitine ratio of 0.63 +/- 0.06 compared with 0.31 +/- 0.16 in trained animals receiving an identical acute bout of exercise, demonstrating that untrained animals utilized a significantly higher percentage of free carnitine reserves during exercise. This study suggests that free carnitine reserves may be reduced during exercise in untrained rats, an effect that has the potential to impair both carbohydrate and fat metabolism during exercise.     

Effect of short-term consumption of a high fat diet on glucose tolerance and insulin sensitivity in the rat

The mechanism by which the consumption of high fat, low carbohydrate diets impair glucose tolerance and decrease insulin sensitivity is poorly understood. In an attempt to clarify this question, intravenous glucose tolerance and insulin action in the liver and skeletal muscle were examined in rats after two weeks feeding of either a high fat (HF: 66% energy as fat) or a low fat (LF: 12% energy as fat) diet. Both diets had a P/S ratio (ratio of polyunsaturated to saturated fat in the diet) of 1.3. The high fat diet resulted in mild impairment of intravenous glucose tolerance. Postprandial glucose levels were elevated in the presence of a sustained insulin response. In vitro insulin-stimulated glucose utilisation was decreased significantly in soleus muscle of HF rats, as indicated by decreased [14C]glucose incorporation into muscle glycogen. In contrast, muscle lipogenesis from glucose was not affected by dietary composition. There was no difference in insulin binding to soleus muscle of HF and LF rats, indicating a dissociation between insulin receptor binding and post-receptor metabolic events. Dietary composition did not influence the incorporation of increasing [14C]glucose loads into muscle glycogen or lipid in vivo. However, the HF diet was associated with reduced incorporation of [14C]glucose into lipids and glycogen in the liver and, to a smaller extent, reduced incorporation into adipose tissue lipids in vivo. These results suggest that the mechanism by which HF diets impaired glucose tolerance was mainly hepatic in origin. Decreased glucose uptake, secondary to reduced glucokinase activity, may result in a reduction in glucose utilisation in the liver.     

Detrimental effects of a high fat diet in early renal injury are ameliorated by fish oil in Han:SPRD-cy rats

Dietary fish oils containing (n-3) fatty acids can modulate renal inflammatory injury. We previously demonstrated that a high fat (HF) diet worsens early renal disease progression in the Han:SPRD-cy rat model of polycystic kidney disease (PKD). Therefore, using HF (20 g/100 g diet) and low fat (LF; 5 g/100 g diet) diets, we compared the effects of menhaden oil (MO), soybean oil (SO) and cottonseed oil (CO) on renal function and histology in male Han:SPRD-cy rats fed the diets for 6 wk in the early stages of renal disease. Overall, rats fed HF compared with those fed LF diets had larger kidneys, more renal fibrosis and lower creatinine clearance (main effects of fat level).Rats fed MO rather than CO and SO diets had significantly lower kidney weights, kidney water content, cyst volumes and serum cholesterol and triglyceride concentrations (main effects of fat type). Rats fed MO diets also had less renal fibrosis than those fed CO diets, but the least fibrosis was in rats fed SO diets. Analysis of simple effects (due to interactions between fat level and type) revealed that HF diets increased renal inflammation in rats fed CO diets, but reduced inflammation was present in those fed SO and MO diets; HF diets also increased compared with LF diets serum urea nitrogen concentrations in rats fed the MO and CO diets, but not the SO diet. These results confirm that high dietary fat worsens early disease progression in this model of renal disease, and further demonstrate that diets with oils containing (n-3) fatty acids ameliorate some of the detrimental effects of a high fat diet.     

Carnitine in the streptozotocin-diabetic rat

Rats made moderately diabetic by streptozotocin (decreased growth without decreased body mass, blood glucose, 588-840 mg/dl) and nondiabetic rats were either fed a purified diet with no measurable carnitine (diet 1), the same diet supplemented with carnitine (44.5 nmol/g food, diet 2), or a closed formula, nonpurified diet (3.3 nmol/g food, diet 3). Levels of total carnitine (free + acyl) were lower (P less than 0.05) in plasma, heart, soleus, extensor digitorum longus and kidney of rats fed diet 1 compared to those fed diet 2. Liver carnitine in diabetic animals was 2.6- to 4.4-fold higher than in nondiabetic animals with urinary carnitine of rats fed diet 1 being four times greater in diabetic than in nondiabetic rats. Other changes in tissue total carnitine were less pronounced. Compared to diabetic rats, insulin-treated diabetics had lower urinary excretion of carnitine and blood glucose levels (P less than 0.05), and tissue carnitine approximating nondiabetic levels. Significant catabolism of carnitine was not found. Apparent carnitine biosynthesis (diet 1) showed no difference between nondiabetic and diabetics (31.5 +/- 1.6 and 31.2 +/- 2.2 nmol/g of body weight per day, respectively) suggesting elevated liver levels resulted from redistribution.     

In vivo evidence for a vitamin B-6 requirement in carnitine synthesis

The purpose of this study was to determine whether there is a requirement for vitamin B-6 (B6) in carnitine synthesis. Rats were fed a B6-deficient (-B6) (0.04 mg pyridoxine [PN]/kg) diet (ad libitum or meal-fed) or a control (+B6) (5.7 mg PN/kg) diet (ad libitum or pair-fed). These diets were fed for 6 wk, then some of the rats were repleted with the +B6 diet for 2 wk. Total acid-soluble carnitine (TCN) and free carnitine (FCN) levels were compared in the plasma, liver, skeletal muscle, heart muscle and urine and rats fed +B6 or -B6 diets. In -B6 rats vs. +B6 rats, TCN levels were significantly lower (P less than 0.05) in the plasma, skeletal muscle, heart muscle and urine, but not in the liver. However, if rats were fasted for 3 d, liver TCN concentration of -B6 rats was significantly lower than that of +B6 rats. After B6-deficient rats were repleted with the +B6 diet, the TCN level in the plasma, liver, skeletal muscle, heart muscle and urine returned to the levels of control rats. Thus, the decrease in TCN and FCN levels with a B6-deficient diet and the increase of these levels after B6 repletion provides evidence for the B6 requirement in the biosynthesis of carnitine.     

Adaptation to a high-fat diet leads to hyperphagia and diminished sensitivity to cholecystokinin in rats

Rats fed high-fat (HF) diets exhibit reduced sensitivity to some peptide satiety signals. We hypothesized that reduced sensitivity to satiety signals might contribute to overconsumption of a high-energy food after adaptation to HF diets. To test this, we measured daily, 3-h intake of a high-energy, high-fat (HHF, 22.3 kJ/g) test food in rats fed either low-fat (LF) or HF, isoenergetic (16.2 kJ/g) diets. During testing, half of each group received the HHF test food (LF/HHF; HF/HHF), whereas the other half received their respective maintenance diet (LF/LF; HF/HF). Rats fed a HF diet ate more of the HHF food during the 3-h testing period than LF-fed rats (HF/HHF = 7.7 +/- 0.3 g vs. LF/HHF = 5.5 +/- 0.2 g; P = 0.003). Rats tested on their own maintenance diets had similar intakes (HF/HF = 3.2 +/- 0.2 g vs. LF/LF = 3.7 +/- 0.3 g), which were lower (P < or = 0.008) than intakes of rats tested on HHF. HHF-tested rats did not differ in body weight by the end of wk 2 of testing. In a subsequent short-term choice preference test, rats exhibited an equal relative preference for HHF irrespective of their maintenance diets (HF = 63.1%, LF = 68.1%, P = 0.29). Finally, we examined the effect of intraperitoneal NaCl or cholecystokinin (CCK)-8 (100 and 250 ng/kg) injection on 1-h food intake. Both doses of CCK significantly suppressed food intake in LF-fed rats but not HF-fed rats. These results demonstrate that chronic ingestion of a HF diet leads to short-term overconsumption of a high-energy, high-fat food compared with LF-fed cohorts, which is associated with a decreased sensitivity to CCK.