Effects of Dietary Fatty Acids and Exercise on Body‐Weight Regulation and Metabolism in Rats

Objective: To assess the interaction of high‐fat diets (HF) made with different dietary fatty acids and exercise on body‐weight regulation, adiposity, and metabolism. Research Methods and Procedures: Male Wistar rats born to dams fed HF diets (40% w/w) made with either fish oil (FO), soybean oil (SO), or palm oil (PO) were fed diets similar to their dams and divided randomly into exercise (EX, swimming) or sedentary control (SD) groups when they were 9 weeks old. EX lasted for 6 weeks. Twenty‐four hours after the last EX bout, fasted rats were killed by decapitation. Chemical analyses and body composition analysis were conducted. Results: The results demonstrated that different fatty acids had different effects on body weight, composition, and metabolism. SO‐fed rats gained the most weight and fat. EX reduced body weight of FO‐ and PO‐fed rats, but SO‐fed rats were still heavier and fatter than other rats. Data from SO‐ and PO‐fed rats suggested that they are insulin resistant and that EX normalized this abnormality. Of the three HF diets used, FO produced the least adverse effects compared with PO and SO. Discussion: Not only the quantity of dietary fat, but also the type of fat used, will produce different effects on body weight and metabolism. EX ameliorates the suggested insulin resistance induced in rats fed either highly saturated or n‐6 polyunsaturated fatty acids. Long‐chain n‐3 polyunsaturated fatty acids, as found in fish oil, are more beneficial than n‐6 polyunsaturated fatty acids when fed in high amounts to rats.     

Growth performance,antioxidant and immune status of Siberian sturgeon (Acipenser baeri) fed diets containing green tea extract and oxidized fish oil

The present study was conducted to determine the effects of dietary oxidized fish oil (OFO) and green tea extract (GTE) on growth performance, antioxidant status and immunological parameters in Siberian sturgeon (Acipenser baeri). Fish weighing about 108 g were randomly assigned to one of nine dietary treatments (control diet without OFO and GTE; diets containing only 25 or 50 mg/kg of GTE; diets containing 50 mg/kg of OFO combined with 0, 25 or 50 mg/kg of GTE; diets containing 100 mg/kg of OFO combined with 0, 25 or 50 mg/kg of GTE) with three replicates of 15 fish stocked in fiberglass tanks using a completely randomized design. Growth performance indices, immunological parameters and antioxidant status of all treatment groups were determined using appropriate assays and compared. Results showed that GTE could enhance growth performance. Feeding GTE alone or combined with the OFO could decrease the hepatosomatic index (HSI) and viscerosomatic index (VSI). Serum lysozyme activity and antibody level were increased significantly by GTE alone or combined with OFO compared to the oxidized diets (p < 0.05). Conversely, lipid peroxidation products and antioxidant enzyme activity were significantly lower in GTE treatment groups and those that received combination of GTE and OFO compared to the oxidized diets. Low‐density lipoprotein (LDL) level in GTE groups was lower than other treatment groups (p < 0.05) however, in OFO treatment groups the LDL level in fish serum was significantly increased in comparison with the other treatment groups (p < 0.05). GTE and OFO alone or combined together significantly decreased the levels of triglyceride and cholesterol compared to the control group (p < 0.05). In conclusion, immunological and biochemical parameters were improved by inclusion of GTE into Siberian sturgeon rancid diets which can protect fish from the oxidative damage challenged by dietary oil oxidation.     

Maternal diets enriched in olive oil regulate lipid metabolism and levels of PPARs and their coactivators in the fetal liver in a rat model of gestational diabetes mellitus

In a rat model of gestational diabetes mellitus (GDM) programmed in the offspring of neonatal streptozotocin-induced (nSTZ) diabetic rats, lipids are accumulated in the fetal liver in a sex-dependent way. Here, we evaluated whether maternal diets enriched in olive oil in rats that will develop GDM ameliorate lipid metabolic impairments in the fetal livers. Pregnant offspring of control and nSTZ diabetic rats (F0) were fed a 6% olive oil-supplemented diet throughout the F1 gestation. We evaluated maternal metabolic parameters as well as lipid content, expression of lipid metabolizing enzymes and protein expression of PLIN2, PPARs and PPAR coactivators in the fetal livers. The offspring of nSTZ diabetic rats developed GDM regardless of the maternal treatment. Hypertriglyceridemia in GDM rats was prevented by the olive oil-enriched maternal treatment. In the livers of male fetuses of GDM rats, the maternal olive oil-supplemented diet prevented lipid overaccumulation and prevented the increase in PPARγ and PPARδ levels. In the livers of female fetuses of GDM rats, the maternal olive oil supplementation prevented the increase in PPARδ levels and the reduction in PGC1α levels, but did not prevent the reduced lipid content. Control and GDM rats showed a reduction of lipid metabolic enzymes in the fetal livers, which was associated with reduced levels of the PPAR coactivators PGC-1α and SRC-1 in males and of SRC-1 in females. These results suggest powerful effects of a maternal olive oil-supplemented diet in the fetal liver, possibly providing benefits in the fetuses and offspring from GDM rats.     

Consumption of soybean or olive oil at recommended concentrations increased the intestinal microbiota diversity and insulin sensitivity and prevented fatty liver compared to the effects of coconut oil

Diets rich in mono or polyunsaturated fats have been associated with a healthy phenotype, but there is controversial evidence about coconut oil (CO), which is rich in saturated medium-chain fatty acids. Therefore, the purpose of the present work was to study whether different types of oils rich in polyunsaturated (soybean oil, SO), monounsaturated (olive oil, OO), or saturated fatty acids (coconut oil, CO) can regulate the gut microbiota, insulin sensitivity, inflammation, mitochondrial function in wild type and PPARα KO mice. The group that received SO showed the highest microbial diversity, increase in Akkermansia muciniphila, high insulin sensitivity and low grade inflammation, The OO group showed similar insulin sensitivity and insulin signaling than SO, increase in Bifidobacterium, increase in fatty acid oxidation and low grade inflammation. The CO consumption led to the lowest bacterial diversity, a 9-fold increase in the LPS concentration leading to metabolic endotoxemia, hepatic steatosis, increased lipogenesis, highest LDL-cholesterol concentration and the lowest respiratory capacity and fatty acid oxidation in the mitochondria. The absence of PPARα decreased alpha diversity and increased LPS concentration particularly in the CO group, and increased insulin sensitivity in the groups fed SO or OO. These results indicate that consuming mono or polyunsaturated fatty acids produced health benefits at the recommended intake but a high concentration of oils (three times the recommended oil intake in rodents) significantly decreased the microbial alpha-diversity independent of the type of oil.     

Activation of peroxisome proliferator-activated receptor-alpha stimulates both differentiation and fatty acid oxidation in adipocytes

Peroxisome proliferator-activated receptor-α (PPARα) is a dietary lipid sensor, whose activation results in hypolipidemic effects. In this study, we investigated whether PPARα activation affects energy metabolism in white adipose tissue (WAT). Activation of PPARα by its agonist (bezafibrate) markedly reduced adiposity in KK mice fed a high-fat diet. In 3T3-L1 adipocytes, addition of GW7647, a highly specific PPARα agonist, during adipocyte differentiation enhanced glycerol-3-phosphate dehydrogenase activity, insulin-stimulated glucose uptake, and adipogenic gene expression. However, triglyceride accumulation was not increased by PPARα activation. PPARα activation induced expression of target genes involved in FA oxidation and stimulated FA oxidation. In WAT of KK mice treated with bezafibrate, both adipogenic and FA oxidation-related genes were significantly upregulated. These changes in mRNA expression were not observed in PPARα-deficient mice. Bezafibrate treatment enhanced FA oxidation in isolated adipocytes, suppressing adipocyte hypertrophy. Chromatin immunoprecipitation (ChIP) assay revealed that PPARα was recruited to promoter regions of both adipogenic and FA oxidation-related genes in the presence of GW7647 in 3T3-L1 adipocytes. These findings indicate that the activation of PPARα affects energy metabolism in adipocytes, and PPARα activation in WAT may contribute to the clinical effects of fibrate drugs.     

PPARα-LXR as a novel metabolostatic signalling axis in skeletal muscle that acts to optimize substrate selection in response to nutrient status

LXR (liver X receptor) and PPARα (peroxisome-proliferator-activated receptor α) are nuclear receptors that control the expression of genes involved in glucose and lipid homoeostasis. Using wild-type and PPARα-null mice fed on an LXR-agonist-supplemented diet, the present study analysed the impact of pharmacological LXR activation on the expression of metabolically important genes in skeletal muscle, testing the hypothesis that LXR activation can modulate PPAR action in skeletal muscle in a manner dependent on nutritional status. In the fed state, LXR activation promoted a gene profile favouring lipid storage and glucose oxidation, increasing SCD1 (stearoyl-CoA desaturase 1) expression and down-regulating PGC-1α (PPARγ co-activator-1α) and PDK4 (pyruvate dehydrogenase kinase 4) expression. PPARα deficiency enhanced LXR stimulation of SCD1 expression, and facilitated elevated SREBP-1 (sterol-regulatory-element-binding protein-1) expression. However, LXR-mediated down-regulation of PGC-1α and PDK4 was opposed and reversed by PPARα deficiency. During fasting, prior LXR activation augmented PPARα signalling to heighten FA (fatty acid) oxidation and decrease glucose oxidation by augmenting fasting-induced up-regulation of PGC-1α and PDK4 expression, effects opposed by PPARα deficiency. Starvation-induced down-regulation of SCD1 expression was opposed by antecedent LXR activation in wild-type mice, an effect enhanced further by PPARα deficiency, which may elicit increased channelling of FA into triacylglycerol to limit lipotoxicity. Our results also identified potential regulatory links between the protein deacetylases SIRT1 (sirtuin 1) and SIRT3 and PDK4 expression in muscle from fasted mice, with a requirement for PPARα. In summary, we therefore propose that a LXR-PPARα signalling axis acts as a metabolostatic regulatory mechanism to optimize substrate selection and disposition in skeletal muscle according to metabolic requirement.     

Prenatal Zinc Deficiency: Influence on Heart Morphology and Distribution of Key Heart Proteins in a Rat Model

The etiology of congenital heart disease is multifactorial, with genetics and nutritional deficiencies recognized as causative agents. Maternal zinc (Zn) deficiency is associated with an increased risk for fetal heart malformations; however, the contributing mechanisms have yet to be identified. In this study, we fed pregnant rats a Zn-adequate diet (ZnA), a Zn-deficient (ZnD), or a restricted amount of Zn adequate diet (RF) beginning on gestation day (GD) 4.5, to examine whether increased cell death and changes in cardiac neural crest cells (NCC) play a role in Zn deficiency-induced heart defects. Fetuses were collected on GD 13.5, 15.5, and 18.5 and processed for GATA-4, FOG-2, connexin-43 (Cx43), HNK-1, smooth muscle α-actin (SMA) and cleaved caspase-3 protein expression. Fetuses from ZnA-fed dams showed normal heart development, whereas fetuses from dams fed with the ZnD diet exhibited a variety of heart anomalies, particularly in the region of the outflow tract. HNK-1 expression was lower than normal in the hearts of GD13.5 and 15.5 ZnD fetuses, particularly in the right atrium and in the distal tip of the interventricular septum. Conversely, Cx43 immunoreactivity was increased throughout the heart in fetuses from ZnD dams compared to fetuses from control dams. The distribution and intensity of expression of SMA, GATA-4, FOG-2, and markers of apoptosis were similar among the three groups. We propose that Zn deficiency induced alterations in the distribution of Cx43 and HNK-1 in fetal hearts contribute to the occurrence of the developmental heart anomalies.     

CCAAT/Enhancer-Binding Protein β is not Affected by Tetrachlorodibenzo-p-dioxin (TCDD) Inhibition of 3T3-L1 Preadipocyte Differentiation

The differentiation of 3T3-L1 preadipocytes is induced by the coordinate activation of trans-acting factors in response to inducers. Depending on the time of treatment, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was effective in inhibiting 3T3-L1 preadipocyte differentiation and the expression of differentiation-dependent trans-acting factors. Based on glycerol-3-phosphate dehydrogenase activity, the differentiation of 3T3-L1 cells was decreased by 70% in cells treated with TCDD before the induction of differentiation, 25% during induction, and not at all after induction. This time-dependent inhibition of cell differentiation by TCDD was correlated with the levels of aryl hydrocarbon receptor (AhR). TCDD treatment decreased the mRNA levels of C/EBPα and PPAR-γ2 but did not affect the mRNA levels of RXRα and RARα. Furthermore, TCDD did not change the mRNA or protein levels of C/EBPβ, which is thought to play a role in inducing C/EBPα and PPARγ2 expression. These results suggest that TCDD inhibited 3T3-L1 preadipocyte differentiation through the AhR pathway, and the change of C/EBPβ mRNA and protein was not involved in reducing mRNA expression of C/EBPα and PPARγ2.     

Troglitazone enhances tamoxifen-induced growth inhibitory activity of MCF-7 cells

Peroxisome proliferator-activated receptor γ (PPARγ) ligands have been identified as a potential source of therapy for human cancers. However, PPARγ ligands have a limitation for breast cancer therapy, since estrogen receptor α (ER_α) negatively interferes with PPARγ signaling in breast cancer cells. Here we show that ER_α inhihits PPARγ transactivity and ER_α-mediated inhibition of PPARγ transactivity is blocked by tamoxifen, an estrogen receptor blocker. The activation of ER_α with 17-β-estradiol blocked PPRE transactivity induced by troglitazone, a PPARγ ligand, indicating the resistance of ER_α-positive breast cancer cells to troglitazone. Indeed, troglitazone inhibited the growth of ER_α-negative MDA-MB-231 cells more than that of ER_α-positive MCF-7 cells. Combination of troglitazone with tamoxifen led to a marked increase in growth inhibition of ER_α-positive MCF-7 cells compared to either agent alone. Our data indicates that troglitazone enhances the growth inhibitory activity of tamoxifen in ER_α-positive MCF-7 cells.     

Molecular characterisation of tumour necrosis factor alpha and its potential connection with lipoprotein lipase and peroxisome proliferator-activated receptors in blunt snout bream (<Emphasis Type="Italic">Megalobrama amblycephala</Emphasis>)

Tumour necrosis factor alpha (TNF-α) is one kind of cytokines which is related to inflammation and lipid metabolism. TNF-α cDNA was cloned from the liver of blunt snout bream (Megalobrama amblycephala) through real-time polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) methods. The full-length cDNA of TNF-α covered 1467 bp, with an open reading frame (ORF) of 723 bp, which encodes 240 amino acids. It possessed the TNF family signature IIIPDDGIYFVYSQ. After the lipopolysaccharide (LPS) challenge test, a graded tissue-specific expression pattern of TNF-α was observed and there was high expression abundance in the kidney, brain and liver. After 8 weeks feeding trial, liver samples, two groups fed with 6% and 11% lipid levels, were collected. The results showed that, for fish fed with high-fat diet, the triglyceride of serum and lipid content of liver were elevated. Furthermore, TNF-α and peroxisome proliferator-activated receptors (PPARα, β) mRNA expression of fish fed 11% lipid diet were significantly up-regulated (p?<?0.05). Lipoprotein lipase (LPL) and PPARγ mRNA expression of fish fed 11% lipid lever diet were significantly decreased compared to those of fish fed 6% (p?<?0.05). The differences between the various expression of related genes in the high and low fat groups demonstrated that TNF-α played a key role in lipid metabolism, which may have an influence on fat metabolism through reducing fat synthesis and strengthening the β-oxidation of fatty acid. These discrepancies warrant further research.     

Maternal omega-3 fatty acids maintained positive maternal lipids and cytokines profile,and improved pregnancy outcomes of C57BL/6 mice

Omega (n)-3 polyunsaturated fatty acids (PUFA) are known to regulate lipid metabolism and inflammation; however, the regulation of maternal lipid metabolism and cytokines profile by n-3 PUFA during different gestation stages, and its impact on fetal sustainability is not known. We investigated the effects of maternal diet varying in n-3 PUFA prior to, and during gestation, on maternal metabolic profile, placental inflammatory cytokines, and fetal outcomes. Female C57BL/6 mice were fed either a high, low or very low (9, 3 or 1% w/w n-3 PUFA) diet, containing n-6:n-3 PUFA of 5:1, 20:1 and 40:1, respectively for two weeks before mating, and throughout pregnancy. Animals were sacrificed prior to mating (NP), and during pregnancy at gestation days 6.5, 12.5 and 18.5. Maternal metabolic profile, placental cytokines and fetal outcomes were determined. Our results show for the first time that a maternal diet high in n-3 PUFA prevented dyslipidemia in NP mice, and maintained the expected lipid profile during pregnancy. However, females fed the very low n-3 PUFA diet became hyperlipidemic prior to pregnancy, and carried this profile into pregnancy. Maternal diet high in n-3 PUFA maintained maternal plasma progesterone and placental pro-inflammatory cytokines profile, and sustained fetal numbers throughout pregnancy, while females fed the low and very-low n-3 PUFA diet had fewer fetuses. Our findings demonstrate the importance of maternal diet before, and during pregnancy, to maintain maternal metabolic profile and fetus sustainability. These findings are important when designing dietary strategies to optimize maternal metabolism during pregnancy for successful pregnancy outcome.     

Dietary fat impacts fetal growth and metabolism: uptake of chylomicron remnant core lipids by the placenta

The fetus requires significant energy for growth and development. Although glucose is a major source of energy for the fetus, other maternal nutrients also appear to promote growth. Thus, the goal of these studies was to determine whether triglyceride-rich remnants are taken up by the placenta and whether maternal dietary lipids, independently of adiposity, can impact fetal growth. To accomplish our first goal, chylomicron particles were duallly labeled with cholesteryl ester and triglycerides. The placenta took up remnant particles/core lipids at rates greater than adipose tissue and skeletal muscle but less than the liver. Although the placenta expresses apoE receptors, uptake of chylomicron remnants and/or core lipids can occur independently of apoE. To determine the impact of dietary lipid on fetal growth, independent of maternal adiposity, females were fed high-fat diets (HFD) for 1 mo; there was no change in adiposity or leptin levels prior to or during pregnancy of dams fed HFD. Fetal masses were greater in dams fed HFD, and mRNA levels of proteins involved in fatty acid oxidation (CPT I, PPARα), but not glucose oxidation (pyruvate kinase) or other regulatory processes (HNF-4α, LXR), were increased with maternal dietary fat. There was also no change in mRNA levels of proteins involved in placental glucose and fatty acid transport, and GLUT1 protein levels in microvillous membranes were similar in placentas of dams fed either diet. Thus, the ability of the placenta to take up chylomicron remnant core lipids likely contributes to accelerated fetal growth in females fed high fat diets.     

Hepatic insulin binding following in utero exposure to ethanol

Insulin binding to liver membranes has been studied in term fetuses of rats fed ethanol-containing liquid diet during pregnancy . Pair-fed and ad libitum-fed controls received liquid diet in which maltose-dextrins were substituted isocalorically for ethanol. Food consumption and body weigh gain of ethanol- imbibing dams were 35% and 70% less than their ad libitum counterparts respectively. Ethanol-fed rats also exhibited less gain in body weight than pair-fed controls despite isocalorically equivalent food intake. The number of live pups was not different among the various groups; however, liver weight of fetuses exposed to ethanol in utero was 47% less than those of the pups of ad libitum control dams and 28% less than those of the offspring of pair-fed control rats. Insulin binding to liver membranes of fetuses exposed to ethanol in utero was lower than that of ad libitum controls but was not significantly different from that of the pair-fed control animals. Average affinity profiles showed a reduction in K at all levels of receptor occupancy in the fetuses of ethanol-fed rats. For fetuses of the pair-fed group, K was reduced only at fractional occupancy below 20% but not at higher fractional occupancy. Because of the similarity of insulin binding in the fetuses of the ethanol-fed rats and their pair-fed counterparts, effects of ethanol on insulin binding cannot account for the reduced hepatic glycogen stores previously reported in term fetuses.     

Progressive, transgenerational changes in offspring phenotype and epigenotype following nutritional transition

Induction of altered phenotypes during development in response to environmental input involves epigenetic changes. Phenotypic traits can be passed between generations by a variety of mechanisms, including direct transmission of epigenetic states or by induction of epigenetic marks de novo in each generation. To distinguish between these possibilities we measured epigenetic marks over four generations in rats exposed to a sustained environmental challenge. Dietary energy was increased by 25% at conception in F0 female rats and maintained at this level to generation F3. F0 dams showed higher pregnancy weight gain, but lower weight gain and food intake during lactation than F1 and F2 dams. On gestational day 8, fasting plasma glucose concentration was higher and β-hydroxybutyrate lower in F0 and F1 dams than F2 dams. This was accompanied by decreased phosphoenolpyruvate carboxykinase (PEPCK) and increased PPARα and carnitine palmitoyl transferase-1 mRNA expression. PEPCK mRNA expression was inversely related to the methylation of specific CpG dinucleotides in its promoter. DNA methyltransferase (Dnmt) 3a2, but not Dnmt1 or Dnmt3b, expression increased and methylation of its promoter decreased from F1 to F3 generations. These data suggest that the regulation of energy metabolism during pregnancy and lactation within a generation is influenced by the maternal phenotype in the preceding generation and the environment during the current pregnancy. The transgenerational effects on phenotype were associated with altered DNA methylation of specific genes in a manner consistent with induction de novo of epigenetic marks in each generation.     

Nucleic acid content and growth of fetal brain, liver and heart during inanition in pigs

Previous investigations have indicated that gilts deprived of dietary intake for periods up to 40 days are capable of maintaining pregnancy and producing offspring of normal body weight. The present experiment was designed to study the effects of inanition during middle or late pregnancy on growth and on protein and nucleic acid content of porcine fetal brain, liver and heart. Gilts were subjected to prolonged inanition (40 days; 0 kcal/day; water only) during either the middle third (Days 30-70, n = 3) or last third (Days 70-110, n = 3) of pregnancy; controls received a full diet (7028 kcal/day) until Day 70 (n = 3) or 110 (n = 3) when all dams were hysterectomized. Inanition during middle or late pregnancy had no detrimental effect on fetal brain development. Brain weight, cell size (protein/DNA ratio) and cell number (total DNA) were similar in all fetuses at Day 70 or 110. RNA concentrations at Days 70 and 110, protein concentration at Day 70 and total protein at Day 110 were higher in fetuses from starved dams than in those from controls, indicating greater protein synthetic activity in fetal brains from nutrient-deprived dams. Prolonged inanition during midpregnancy had only a limited effect on fetal liver and heart. Only liver RNA concentration and content at Day 70 differed in fetuses from starved dams; however, 40 days inanition during late gestation had marked detrimental effects. Liver weight, cell size and cell number were reduced in inanition as compared with controls by Day 110.(ABSTRACT TRUNCATED AT 250 WORDS)