Effect of polyunsaturated fatty acids in obese mice

Genetically obese (ob/ob) mice display a variety of metabolic differences from lean litter mates. In the obese state, fatty acid desaturation-elongation in brown adipose tissue mitochondria is apparently altered, resulting in differences in membrane fatty acid composition. This change in membrane lipid environment appears to influence GDP binding and there-fore the activity of the proton conductance pathway associated with regulation of energy expenditure in these animals. In liver, binding of insulin to the nuclear membrane is increased by feeding a high polyunsaturated/saturated (P/S) diet fat. Consumption of a high P/S diet decreased mRNA levels for fatty acid synthase, acetyl-CoA carboxylase, malic enzyme, and pyruvate kinase in obese and lean animals. Expression of mRNA for these lipogenic enzymes was higher in obese animals and suggests that obese mice may be resistant to polyunsaturated fatty acid feedback control of gene expression.     

Differential effects of obesity on acetoacetyl‐CoA synthetase gene in rat adipose tissues

In order to investigate the physiological role of the ketone body‐utilizing enzyme acetoacetyl‐CoA synthetase (AACS) in obesity‐induced metabolic derangements, we examined the mRNA levels of AACS and other lipogenic enzymes in white adipose tissues (WAT) in the different types of obesity, genetic (Zucker fatty) and high‐fat diet (HFD)‐induced obesity. Plasma ketone body levels and gene expression levels of AACS and acetyl‐CoA carboxylase‐2 (ACC‐2) in WAT were lower in Zucker fatty rats than in lean rats. On the contrary, in HFD‐induced obese rats, these levels were increased. Moreover, AACS mRNA expression was affected only in subcutaneous WAT by each type of obesity. Our data suggest that AACS mRNA expression is related to obesity‐induced alteration of plasma ketone body levels, and that AACS in subcutaneous WAT plays an important role in the regulation of lipogenesis and ketone body utilization in various obese conditions.     

Dietary Saury Oil Reduces Hyperglycemia and Hyperlipidemia in Diabetic KKAy Mice and in Diet-Induced Obese C57BL/6J Mice by Altering Gene Expression

We investigated the effect of saury oil on the alleviation of metabolic syndrome in mice. Saury oil contains 18% (w/w/) n-3 polyunsaturated fatty acids (n-3 PUFA) and 35% (w/w) monounsaturated fatty acids (MUFA). Diabetic KKAy mice were fed a 10% soybean oil diet (control) or a 10% saury oil diet for 4 weeks, and diet-induced obese C57BL/6J mice were fed a high-fat diet containing 32% lard (control) or 22% lard plus 10% saury oil for 6 weeks. After the intervention periods, the levels of glucose, insulin and lipids in plasma had decreased significantly for the saury oil diet group, and insulin sensitivity had improved. These favorable changes may be attributed to the increased adiponectin and decreased TNFα and resistin levels in plasma. The saury oil diet also resulted in downregulated expression of the lipogenic genes (SREBP-1, SCD-1, FAS, and ACC) as well as upregulation of the fatty acid oxidative gene, CPT-1, and the energy expenditure-related genes (PGC1α and PGC1β) in white adipose tissue for the diet-induced obese C57BL/6J mice. An increase in n-3 PUFA levels and the concomitant decrease in the n-6/n-3 PUFA level ratio in serum, white adipose tissue, and liver with a saury oil diet are likely to be involved in the beneficial changes to the metabolic indicators. MUFA may also play a positive role in remodeling lipid composition. Based on these mice models, our results suggest a potential use for saury oil for improving metabolic abnormalities.     

Cinnamomum camphora Seed Kernel Oil Improves Lipid Metabolism and Enhances β3-Adrenergic Receptor Expression in Diet-Induced Obese Rats

The effects of dietary Cinnamomum camphora seed kernel oil (CCSKO) containing medium‐chain triacylglycerols on lipid metabolism and mRNA and protein expression of β‐3 adrenergic receptor in adipose tissue were studied in diet‐induced obese rats. High fat food‐induced obese rats were randomly divided into CCSKO group, Lard group, Soybean oil (SOY) group and naturally restoring group (n = 10). Rats fed with low fat food were used as a normal control group. Significant decreases in body mass and abdominal fat mass/body mass after 12 weeks were found in CCSKO group as compared with Lard and SOY groups (p < 0.05). Levels of blood total cholesterol (TC), triglyceride, free fatty acid, fasting insulin and insulin resistance in the CCSKO group were decreased significantly, and noradrenaline level and insulin sensitivity index in the CCSKO group were significantly higher than other groups. Meanwhile liver TC and triglyceride levels in the CCSKO group were also decreased markedly. Expression levels of β3‐adrenergic receptor mRNA and protein were higher in CCSKO group than in Lard and SOY groups. These results suggest that CCSKO may contribute to reduction of the body fat mass, promote lipid metabolism and up‐regulate β3‐adrenergic receptor expression in high fat diet‐induced obese rats.     

Liver-Specific Pyruvate Dehydrogenase Complex Deficiency Upregulates Lipogenesis in Adipose Tissue and Improves Peripheral Insulin Sensitivity

The pyruvate dehydrogenase complex (PDC) plays a critical role in lipid synthesis and glucose homeostasis in the fed and fasting states. The central role of the liver in the maintenance of glucose homeostasis has been established by studying changes in key enzymes (including PDC) and the carbon-flux via several pathways under different metabolic states. In the present study we have developed a murine model of liver-specific PDC deficiency using Cre-loxP technology to investigate its consequences on lipid and carbohydrate metabolism. There was no incorporation of glucose-carbon into fatty acids by liver in vitro from liver-specific Pdha1 knockout (L-PDHKO) male mice due to absence of hepatic PDC activity. Interestingly, there was a compensatory increase in lipogenic capacity in epididymal adipose tissue from L-PDHKO mice. Both fat and lean body mass were significantly reduced in L-PDHKO mice, which might be explained by an increase in total energy expenditure compared with wild-type littermate mice. Furthermore, both liver and peripheral insulin sensitivities measured during a hyperinsulinemic-euglycemic clamp were improved in L-PDHKO mice. The findings presented here demonstrate (i) the indispensable role of PDC for lipogenesis from glucose in liver and (ii) specific adaptations in lipid and glucose metabolism in the liver and adipose tissue to compensate for loss of PDC activity in liver only.     

Black Raspberry Seed Oil Improves Lipid Metabolism by Inhibiting Lipogenesis and Promoting Fatty‐Acid Oxidation in High‐Fat Diet‐Induced Obese Mice and db/db Mice

The objective of this study was to evaluate the beneficial effect of α‐linolenic acid‐rich black raspberry seed (BRS) oil on lipid metabolism in high‐fat diet (HFD)‐induced obese and db/db mice. Five‐week‐old C57BL/6 mice were fed diets consisting of 50% calories from lard, 5% from soybean, and 5% from corn oil (HFD), or 50% calories from lard and 10% from BRS oil (HFD + BRS oil diet) for 12 weeks. Six‐week‐old C57BL/KsJ‐db/db mice were fed diets consisting of 16% calories from soybean oil (standard diet), 8% from soybean, and 8% from BRS oil, or 16% from BRS oil for 10 weeks. The BRS oil diets lowered the levels of triacylglycerol, nonesterified fatty acids, and total cholesterol in serum and liver of both of the obese and db/db mice as compared with the HFD and standard diet, respectively. mRNA levels of lipogenesis markers including cluster of differentiation 36, fatty‐acid‐binding protein 1, sterol regulatory element binding protein 1c, fatty‐acid synthase, and solute carrier family 25 member 1 in the liver of the BRS oil groups were lower than those in the liver of the HFD and standard groups in the obese and db/db mice, respectively. On the other hand, fatty‐acid oxidation markers including carnitine palmitoyltransferase 1A, acyl‐CoA dehydrogenase, hydroxylacyl‐CoA dehydrogenase α, and acyl‐CoA oxidase in the liver of the BRS oil groups were higher than those in the liver of the HFD and standard groups in the obese and db/db mice, respectively. Peroxisome proliferator‐activated receptor α mRNA and protein levels increased in the liver and epididymal adipose tissue of the obese and db/db mice fed BRS oil compared with HFD and standard diet, respectively. BRS oil might improve lipid metabolism by inhibiting lipogenesis and promoting fatty‐acid oxidation in HFD‐induced obese and db/db mice.     

Fatty Acid de Novo Synthesis in Adult Intrauterine Growth-Restricted Offspring,and Adult Male Response to a High Fat Diet

Intrauterine growth restriction (IUGR) with rapid catch‐up growth leads to adult obesity and insulin resistance. We have previously shown that IUGR male rats demonstrated increased de novo fatty acid synthesis in the subcutaneous (SC) fat, but not the visceral fat, during the nursing period prior to the onset of obesity. Young IUGR females do not exhibit the same increase. We further hypothesized that in male IUGR offspring, de novo synthesis is a programmed intrinsic effect that persists to adulthood and does not suppress in response to a high fat diet. We measured fatty acid de novo synthesis in IUGR adult males (6 months) using deuterium‐enriched drinking water as a stable isotope tracer, then further studied the response after consumption of an isocaloric high fat diet. Baseline de novo synthesis in adult females was also studied at age 9 months. Males demonstrated increased baseline de novo synthesis in both SC fat and visceral fat. Correspondingly, SC and visceral fat protein expression of lipogenic enzymes acetyl‐coA carboxylase‐α (ACCα) and fatty acid synthase were upregulated. After the isocaloric high fat diet, de novo synthesis was suppressed such that no differences remained between the two groups, although, IUGR SC fat demonstrated persistently increased lipogenic protein expression. In contrast, de novo synthesis among adult females is not impacted in IUGR. In conclusion, enhancement of male IUGR SC fat de novo synthesis appears to be an early consequence of metabolic programming, whereas enhancement in visceral fat appears to be a later consequence.     

Different Ratios of Corn and Coconut Oil Blends in High-Fat Diets Influence Fat Deposition without Altering Metabolic Biomarkers in Male Rats

The effects of high-fat diets with the recommended dietary linoleic acid (LA) intake levels on health outcomes have not been studied extensively. This study investigated the effects of high-fat diets containing different weight ratios of coconut and corn oil with LA levels of <1.00% of energy (very low LA), 2.80% of energy (low LA), 5.80% of energy (moderate LA), and 9.70% of energy (high LA) on fat deposition and selected metabolic biomarkers of male Sprague-Dawley rats. Their initial and terminal body weights are recorded. Blood, adipose tissue, and liver samples are obtained for analysis after an 8-week feeding intervention. Compared with the very low-LA diet, the high-LA diet resulted in higher body weight gain and epididymal fat deposition. No significant differences are observed in liver-to-body weight ratio, blood glucose, visfatin, and leptin levels between the test diets. Serum tumor necrosis factor-alpha (TNF-α), insulin, and C-peptide levels do not significantly increase with the increase in dietary LA levels. High-LA diet results in higher LA levels in the liver and adipose tissue. It is concluded that a high-fat diet containing high LA levels induced body weight gain and epididymal fat deposition in rats but has no effect on selected metabolic biomarkers. Practical applications: Linoleic acid (LA) (C18:2) plays an important role as one of the nutritional elements to meet the daily essential fatty acid requirements. However, a full understanding is perplexed by the various ways that LA can be included in the diet when there is a recommendation to substitute saturated fatty acid (SFA), trans- or n-3 fatty acids intake. The data provide additional findings on the effects of excessive dietary intake of LA (C18:2) on fat deposition when different levels of SFAs are replaced.     

Protective Effects of Voluntary Exercise on Hepatic Fat Accumulation Induced by Dietary Restriction in Zucker Fatty Rats

Weight control based on dietary restriction (DR) alone can cause lipid metabolic failure and progression to fatty liver. This study aimed to investigate the effect of exercise on preventing DR-induced hepatic fat accumulation in Zucker fatty (ZF) rats by focusing on the relationship between adipose tissue lipolysis and hepatic fat uptake. Six-week-old male ZF rats were randomly assigned to obese, DR, or DR with exercise (DR + Ex) groups. The DR and DR + Ex groups were fed a restricted diet, with the latter also undergoing voluntary exercise. After 6 weeks, hepatic fat accumulation was observed in the DR group, whereas intrahepatic fat was markedly reduced in the DR + Ex group. Compared with the obese (Ob) group, the DR group exhibited 2.09-fold expression of hepatic fatty acid translocase (FAT)/CD36 proteins (p < 0.01) and 0.14-fold expression of hepatic fatty acid-binding protein (FABP)1 (p < 0.01). There were no significant differences between the DR + Ex group and the Ob group. FAT/CD36 and hepatic triglyceride (TG) expression levels were strongly positively correlated (r = 0.81, p < 0.001), whereas there was a strong negative correlation between FABP1 and hepatic TG expression levels (r = −0.65, p < 0.001). Our results suggest that hepatic fat accumulation induced by DR in ZF rats might be prevented through exercise-induced modifications in FAT/CD36 and FABP1 expression.     

Medium chain triglyceride in early life: Effects on growth of adipose tissue

Effects of feeding early in life a diet high in either long chain (LCT) or medium chain triglyceride (MCT) were studied on the development of adipose tissue in post-weanling rats. The diets were similar in calorie distribution and identical in nutrients except for type of fat. The caloric distribution of the two diets by percent was LCT (corn oil)/protein/carbohydrate, 70/18/12 and MCT/corn oil/protein/carbohydrate, 66/4/18/12. Male littermates with less than 5% weight difference were pair-fed the two diets randomly at age 18–20 days. One-fourth of the rats were killed at 10, 16, 22 and 28 weeks of age and analyzed for adipose depots and adipose tissue cellularity. Results showed that the LCT-fed rats were significantly heavier, with larger epididymal, retroperitoneal, omental and subcutaneous fat pads than the respective pair-fed MCT rats. Also, LCT-fed rats had larger size and number of adipocytes than MCT-fed littermates. It is concluded that the type of fat in the diet, namely LCT or MCT, when fed early in life can influence the development of adipose tissue. MCT appears less lipogenic than LCT. The mechanism for the diminished adiposity of MCT-fed rats is related to extensive oxidation of MCT and its enhancement of thermogenesis leading to lessened energy efficiency. Presented at the symposium on “Specialty Lipids and Their Biofunctionality” at the annual meeting of the American Oil Chemists' Society, Philadelphia, May 1985.     

Fatty acid composition of triglycerides from adipose tissue transplanted between obese and lean mice

The subcutaneous adipose tissue of genetically obese mice (ob/ob) differs from that of lean litter-mates not only by virtue of its larger cells but also in its fatty acid composition; it contains a higher proportion of palmitoleic acid and a lower proportion of linoleic acid. To determine whether these differences in fatty acid composition were inherent in fat cells, subcutaneous adipose tissue from obese and lean mice was transplanted under the kidney capsules of lean and obese host mice and the fatty acid composition of the neutral lipids of the graft and of the host perirenal and subcutaneous fat was determined 1 or 2 months later. The fatty acid composition of grafts from lean donors in obese mice resembled that of the perirenal adipose tissue of the obese hosts after 1 month, with a lower proportion of linoleic acid and a higher proportion of palmitoleic acid than in lean mice. Grafts from obese mice in lean mice had fatty acid compositions which were either unchanged, partially changed or which completely resembled that of the host. The use of grafts prelabeled by feeding the donor margaric acid indicated that total lack of fatty acid turnover, rather than selective metabolic processes, was responsible for the failure of some grafts from obese mice in lean mice to acquire the fatty acid composition of the perirenal adipose tissue of the host.     

Hepatic contribution to newly made fatty acids in adipose tissue in rats and inhibition of hepatic and extrahepatic lipogenesis from glucose by dietary corn oil

We have reexamined an earlier rat study in which the authors concluded that 60 min after [U-¹⁴C]-glucose injection half of labeled fatty acids found in adipose tissue had been made in liver and then transported to the adipose tissue. We have shown that even under conditions in which the lipogenic role of the liver is optimized (fed-refed rats on a fat-free, high-carbohydrate diet), almost none of the labeled fatty acids found in adipose tissue of rats 60 min after they were fed a labeled glucose test meal was derived from the liver. This conclusion was based experimentally on (a) the use of the blocking agent Triton WR 1339 to measure the total labeled triglyceride fatty acids (TGFA) synthesized and secreted by the liver in 60 min and (b) comparison of plasma TGFA-¹⁴C data with radioactivity found in liver and in adipose tissue in 60 min. Without using Triton WR 1339, mathematical analysis of plasma TGFA-¹⁴C following the glucose test-meal leads one to the same conclusion: 97% of¹⁴C-labeled fatty acids found in adipose tissue at 60 min was made in situ. Additional studies in rats established that the source of error in the earlier studies was an incorrect assumption that dietary corn oil could inhibit hepatic lipogenesis from glucose C without inhibiting fatty acid synthesis in adipose tissue. In our studies, 10% corn oil inhibited equally both hepatic and adipose tissue fatty acid synthesis from glucose C under conditions that precluded any significant transport of labeled TGFA-¹⁴C from liver to adipose tissue.     

Increased Serum Level of Cyclopropaneoctanoic Acid 2-Hexyl in Patients with Hypertriglyceridemia-Related Disorders

We recently reported the presence of various cyclopropane fatty acids—among them, cyclopropaneoctanoic acid 2‐hexyl—in the adipose tissue of obese women. The aim of this study was to verify whether the presence of cyclopropaneoctanoic acid 2‐hexyl in human serum was associated with obesity or chronic kidney disease (both being related to dyslipidemia), and to find potential associations between the serum level of this compound and specific markers of the these conditions. The serum concentration of cyclopropaneoctanoic acid 2‐hexyl was determined by gas chromatography–mass spectrometry (GC–MS) in non‐obese controls, obese patients, obese patients after a 3‐month low‐calorie diet, and individuals with chronic kidney disease. Obese patients and those with chronic kidney disease presented with higher serum levels of cyclopropaneoctanoic acid 2‐hexyl than controls. Switching obese individuals to a low‐calorie (low‐lipid) diet resulted in a reduction in this fatty acid concentration to the level observed in controls. Cyclopropaneoctanoic acid 2‐hexyl was also found in foods derived from animal fat. Serum concentrations of triacylglycerols in the analyzed groups followed a pattern similar to that for serum cyclopropaneoctanoic acid 2‐hexyl, and these variables were positively correlated with each other among the studied groups. Patients with hypertriglyceridemia‐related conditions presented with elevated serum levels of cyclopropaneoctanoic acid 2‐hexyl. Our findings suggest that its high serum level is related to high serum triacylglycerol concentrations rather than to body mass or BMI.     

High Glucose and Liver Fatty Acid Binding Protein Gene Ablation Differentially Impact Whole Body and Liver Phenotype in High-Fat Pair-Fed Mice

Ad libitum-fed diets high in fat and carbohydrate (especially fructose) induce weight gain, obesity, and nonalcoholic fatty liver disease (NAFLD) in humans and animal models. However, interpretation is complicated since ad libitum feeding of such diets induces hyperphagia and upregulates expression of liver fatty acid binding protein (L-FABP)—a protein intimately involved in fatty acid and glucose regulation of lipid metabolism. Wild-type (WT) and L-fabp gene ablated (LKO) mice were pair-fed either high-fat diet (HFD) or high-fat/high-glucose diet (HFGD) wherein total carbohydrate was maintained constant but the proportion of glucose was increased at the expense of fructose. In LKO mice, the pair-fed HFD increased body weight and lean tissue mass (LTM) but had no effect on fat tissue mass (FTM) or hepatic fatty vacuolation as compared to pair-fed WT counterparts. These LKO mice exhibited upregulation of hepatic proteins in fatty acid uptake and cytosolic transport (caveolin and sterol carrier protein-2), but lower hepatic fatty acid oxidation (decreased serum β-hydroxybutyrate). LKO mice pair-fed HFGD also exhibited increased body weight; however, these mice had increased FTM, not LTM, and increased hepatic fatty vacuolation as compared to pair-fed WT counterparts. These LKO mice also exhibited upregulation of hepatic proteins in fatty acid uptake and cytosolic transport (caveolin and acyl-CoA binding protein, but not sterol carrier protein-2), but there was no change in hepatic fatty acid oxidation (serum β-hydroxybutyrate) as compared to pair-fed WT counterparts.     

Dietary fats and energy levels differently affect tissue lipogenic enzyme activity in finishing pigs

The aim of the present study was to determine the relationship between high and low digestible energy levels (9.5 vs. 15.4 MJ ME/kg) and either tallow or soy oil supplementation (5%rpar; on lipogenic activities and fatty acid profile of the backfat tissue outer layer and liver tissue in finishing pigs. Twenty Large White pigs averaging 30 (initial) to 106 kg (final) live weight were allocated into four dietary groups and fed the diets ad libitum. The lipid content and fatty acid composition of the tissues were determined and glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme (ME), and fatty acid synthase (FAS) activity were measured. Growth performance and carcass measurements were affected by the dietary energy levels but not by the fat sources. Lipid deposition rate of animals fed the low energy diets was lowered regardless whether tallow or soy oil was supplemented. Unlike lipid deposition, fatty acid profile was influenced by both dietary factors. Pigs fed the low energy diet supplemented with soy oil exhibited the lowest level of saturated (P<0.001), monounsaturated (P<0.001), and the highest level of polyenic fatty acids in the backfat, the opposite was the case for the pigs fed the high energy diet supplemented with beef tallow. The fatty acid profile of the adipose tissue of animals fed the other two diets were intermediate, but clear distinction of the profile due to diets was visible. Independent of dietary treatments, lipogenic activities were up to 10 times higher in the backfat than in the liver. G6PDH activity was higher (P<0.05) due to high energy diet, whereas the activities of ME and FAS were not affected. Animals fed the high energy diet either supplemented with tallow or soy oil exhibited higher ME activity lpar;P<0.05) in the backfat, without any effects on G6PDH activity. In contrast, dietary fat sources affected the FAS activity, with lower activity lpar;P<0.05) exhibited in the backfat of animals fed the soy oil diets. The present results indicate that dietary manipulation, which change the flux through the pathway of lipogenesis and pentose-phosphate must affect differently the activities of the involved enzymes. The effect of the dietary energy level was stronger and overwhelmed the inducing effect of the PUFA on the activities of the collateral enzymes. In contrast the immediately involved lipogenic enzyme FAS responded more to dietary PUFA stimulation than to the energy supply.     

Brown adipose tissue triacylglycerol fatty acids of obese and lean mice: In situ and in transplants

The triacylglcyerols of white adipose tissue (WAT) from animals with high rates of lipogenesis, such as obese hyperglycemic mice or hypothalamically lesioned rats, contain high proportions of palmitoleic acid (16∶1) and low proportions of linoleic acid (18∶2). These differences appear to result from dilution of dietary 18∶2 by synthesized fatty acids, particularly 16∶1. To test this we have investigated the triacylglycerol fatty acid composition of brown and white adipose tissue of lean and obese mice, as brown adipose tissue (BAT) has a higher lipogenic rate than WAT and lipogenesis is faster in obese than in lean mice. Between three and eight weeks of age the proportions of fatty acids in the tissues changed, with a marked fall in milk-derived lauric and myristic acids. From 8 to 16 weeks they were more stable and the proportions of 16∶1 and 18∶2 in the different tissues were as expected, with the highest and lowest proportions, respectively, in BAT from obese mice. When BAT from obese mice was transplanted under the kidney capsule of lean mice, or vice versa, for one month, the fatty acid composition of the grafts changed toward that of the host BAT. The proportions of 18∶2 and, to a lesser extent, 16∶1 were slightly higher in the grafts than in the hosts but since this also occurred in lean-to-lean and obese-to-obese grafts it was probably a transplantation artifact. Overall, the results confirm than the physiological environment, rather than the source of the adipose tissue, is the major determinant of its fatty acid composition.