Defining a relationship between dietary fatty acids and the cytochrome P450 system in a mouse model of fatty liver disease.

Liver-specific ablation of cytochrome P450 reductase in mice (LCN) results in hepatic steatosis that can progress to steatohepatitis characterized by inflammation and fibrosis. The specific cause of the fatty liver phenotype is poorly understood but is hypothesized to result from elevated expression of genes encoding fatty acid synthetic genes. Since expression of these genes is known to be suppressed by polyunsaturated fatty acids, we performed physiological and genomics studies to evaluate the effects of dietary linoleic and linolenic fatty acids (PUFA) or arachidonic and decosahexaenoic acids (HUFA) on the hepatic phenotypes of control and LCN mice by comparison with a diet enriched in saturated fatty acids. The dietary interventions with HUFA reduced the fatty liver phenotype in livers of LCN mice and altered the gene expression patterns in these livers to more closely resemble those of control mice. Importantly, the expression of genes encoding lipid pathway enzymes were not different between controls and LCN livers, indicating a strong influence of diet over POR genotype. These analyses highlighted the impact of POR ablation on expression of genes encoding P450 enzymes and proteins involved in stress and inflammation. We also found that livers from animals of both genotypes fed diets enriched in PUFA had gene expression patterns more closely resembling those fed diets enriched in saturated fatty acids. These results strongly suggest only HUFA supplied from an exogenous source can suppress hepatic lipogenesis.

Hypocholesterolemic effect of Nostoc commune var. sphaeroides Kützing, an edible blue-green alga.

BACKGROUND: Intake of an edible blue-green alga Nostoc commune var. sphaeroides Kützing (N. Commune) has been shown to lower plasma total cholesterol concentration, but the mechanisms behind the hypocholesterolemic effect have not been elucidated. AIM OF THE STUDY: To elucidate the mechanisms underlying the cholesterol-lowering effect of N. commune in mice. METHODS: Male C57BL/6J mice were fed the AIN-93 M diet supplemented with 0 or 5% (wt/wt) dried N. Commune for 4 weeks. Lipid levels in the plasma and liver, intestinal cholesterol absorption and fecal sterol excretion were measured. Expression of hepatic and intestinal genes involved in cholesterol metabolism was evaluated by quantitative realtime PCR. RESULTS: N. commune supplementation significantly reduced total plasma cholesterol and triglyceride concentrations by approximately 20% compared to controls. Intestinal cholesterol absorption was significantly decreased, while fecal neutral sterol output was significantly increased in N. commune-fed mice. mRNA levels of the cholesterol transporters such as Niemann Pick C1 Like 1, scavenger receptor class B type 1, ATP-binding cassette transporters G5 and A1 in small intestine were not significantly different between two groups. Hepatic lipid contents including total cholesterol, triglyceride and free cholesterol in N. commune-fed mice were not significantly altered. However, the expression of cholesterol modulating genes including sterol regulatory element binding protein-2 and 3-hydroxy-3-methylglutaryl coenzyme A reductase were significantly increased in mice fed N. commune. CONCLUSIONS: N. commune supplementation exerted a hypocholesterolemic effect in mice, largely in part, by reducing intestinal cholesterol absorption and promoting fecal neutral sterol excretion.

The Short-Term Effects of Lying, Sitting and Standing on Energy Expenditure in Women.

The deleterious health effects of too much sitting have been associated with an increased risk for overweight and obesity. Replacing sitting with standing is the proposed intervention to increase daily energy expenditure (EE). The purpose of this study was to determine the short-term effects of lying, sitting, and standing postures on EE, and determine the magnitude of the effect each posture has on EE using indirect calorimetry (IC). Twenty-eight healthy females performed three separate positions (lying, sitting, standing) in random order. Inspired and expired gases were collected for 45-minutes (15 minutes for each position) using breath-by-breath indirect calorimetry. Oxygen consumption (VO2) and carbon dioxide production (VCO2) were measured to estimate EE. Statistical analyses used repeat measures ANOVA to analyze all variables and post hoc t-tests. Based on the ANOVA the individual, time period and order term did not result in a statistically significant difference. Lying EE and sitting EE were not different from each other (P = 0.56). However, standing EE (kcal/min) was 9.0 % greater than lying EE (kcal/min) (P = 0.003), and 7.1% greater than sitting EE (kcal/min) (P = 0.02). The energetic cost of standing was higher compared to lying and sitting. While this is statistically significant, the magnitude of the effect of standing when compared to sitting was small (Cohen's d = 0.31). Short-term standing does not offer an energetic advantage when compared to sitting.

The Effectiveness of MyPlate and Paleolithic-based Diet Recommendations, both with and without Exercise, on Aerobic Fitness, Muscular Strength and Anaerobic Power in Young Women: A Randomized Clinical Trial.

The purpose of this study was to evaluate the effectiveness of recommending a MyPlate or a Paleolithic-based diet, both with and without exercise, on aerobic fitness, strength, and anaerobic power over eight weeks. Participants (n=20) were randomized to one of four groups, (1) a MyPlate diet (MP), (2) Paleolithic-based diet (PD), (3) MyPlate and exercise (MP + Ex), and (4) Paleolithic-based diet and exercise (PD + Ex). The exercise included two days of unsupervised aerobic and resistance exercise. At baseline and final, absolute and relative peak oxygen consumption (absVO2peak and relVO2peak), anaerobic power, and upper and lower body strength were determined. Data were analyzed using repeated measures two-way analysis of variance (ANOVA). The ANOVA indicated that there was no significant interaction between time point (TP)*diet (D)*exercise (Ex) for all variables except relVO2peak (p = 0.016). The MP + Ex group (Δ+4.4 ml·kg-1·min-1) had a greater change from baseline compared to the MP group (Δ-2.7 ml·kg-1·min-1, p = 0.002), and PD + Ex group (Δ-0.3 ml·kg-1·min-1, p = 0.03). The results suggest recommending a MyPlate diet with both aerobic and resistance training are effective at improving aerobic fitness when compared to PD recommendations with exercise, although these conclusions may be confounded by low compliance to exercise recommendations.

Lentil ( Lens culinaris Medikus) Diet Affects the Gut Microbiome and Obesity Markers in Rat.

Lentil, a moderate-energy high-protein pulse crop, provides significant amounts of essential nutrients for healthy living. The objective of this study was to determine if a lentil-based diet affects food and energy intake, body weight, percent body fat, liver weight, and body plasma triacylglycerols (TGs) as well as the composition of fecal microbiota in rats. A total of 36 Sprague-Dawley rats were treated with either a standard diet, a 3.5% high amylose corn starch diet, or a 70.8% red lentil diet for 6 weeks. By week 6, rats fed the lentil diet had significantly lower mean body weight (443 ± 47 g/rat) than those fed the control (511 ± 51 g/rat) or corn (502 ± 38 g/rat) diets. Further, mean percent body fat and TG concentration were lower, and lean body mass was higher in rats fed the lentil diet than those fed the corn diet. Fecal abundance of Actinobacteria and Bacteriodetes were greater in rats fed the lentil or corn starch diets than those fed the control diet. Fecal abundance of Firmicutes, a bacterial phylum comprising multiple pathogenic species, decreased in rats fed the lentil and high-amylose corn starch diets vs the control diet. The lentil-based diet decreased body weight, percent body fat, and plasma triacylglycerols in rats and suppressed intestinal colonization by pathogens.

Food Ingredients That Inhibit Cholesterol Absorption.

Cholesterol is a vital component of the human body. It stabilizes cell membranes and is the precursor of bile acids, vitamin D and steroid hormones. However, cholesterol accumulation in the bloodstream (hypercholesterolemia) can cause atherosclerotic plaques within artery walls, leading to heart attacks and strokes. The efficiency of cholesterol absorption in the small intestine is of great interest because human and animal studies have linked cholesterol absorption with plasma concentration of total and low density lipoprotein cholesterol. Cholesterol absorption is highly regulated and influenced by particular compounds in the food supply. Therefore, it is desirable to learn more about natural food components that inhibit cholesterol absorption so that food ingredients and dietary supplements can be developed for consumers who wish to manage their plasma cholesterol levels by non-pharmacological means. Food components thus far identified as inhibitors of cholesterol absorption include phytosterols, soluble fibers, phospholipids, and stearic acid.

Approximate Time to Steady-state Resting Energy Expenditure Using Indirect Calorimetry in Young, Healthy Adults.

Indirect calorimetry (IC) measurements to estimate resting energy expenditure (REE) necessitate a stable measurement period or steady state (SS). There is limited evidence when assessing the time to reach SS in young, healthy adults. The aims of this prospective study are to determine the approximate time to necessary reach SS using open-circuit IC and to establish the appropriate duration of SS needed to estimate REE. One hundred young, healthy participants (54 males and 46 females; age = 20.6 ± 2.1 years; body weight = 73.6 ± 16.3 kg; height 172.5 ± 9.3 cm; BMI = 24.5 ± 3.8 kg/m2) completed IC measurement for approximately 30 min while the volume of oxygen (VO2) and volume of carbon dioxide (VCO2) were collected. SS was defined by variations in the VO2 and VCO2 of ≤10% coefficient of variation (%CV) over a period of five consecutive minutes. The 30-min IC measurement was divided into six 5-min segments, such as S1, S2, S3, S4, S5, and S6. The results show that SS was achieved during S2 (%CV = 6.81 ± 3.2%), and the %CV continued to met the SS criteria for the duration of the IC measurement (S3 = 8.07 ± 4.4%, S4 = 7.93 ± 3.7%, S5 = 7.75 ± 4.1%, and S6 = 8.60 ± 4.6%). The current study found that in a population of young, healthy adults the duration of the IC measurement period could be a minimum of 10 min. The first 5-min segment was discarded, while SS occurred by the second 5-min segment.

Unsaturated fatty acids repress the expression of ATP-binding cassette transporter A1 in HepG2 and FHs 74 Int cells.

Adenosine triphosphate-binding cassette transporter A1 (ABCA1) plays a critical role in the formation and metabolism of high-density lipoproteins (HDLs). Adenosine triphosphate-binding cassette transporter A1 in the liver and small intestine, in particular, accounts for approximately 90% of plasma HDL cholesterol. Therefore, any alterations in the hepatic and intestinal expression of ABCA1 could have a large impact on HDL biogenesis. We tested the hypothesis that ABCA1 expression is regulated differentially by different types of fatty acids in the liver and small intestine. Human hepatoma HepG2 and human small intestine epithelial FHs 74 Int cells were used as an in vitro model. Cells were incubated with saturated and unsaturated fatty acids in the presence or absence of T0901317, a synthetic agonist of liver X receptor. Unsaturated fatty acids decreased ABCA1 protein levels at 100 µmol/L of concentration regardless of the agonist with a minimal effect on messenger RNA abundance. Incubation of HepG2 and FHs 74 Int cells with rottlerin, a protein kinase C δ (PKCδ) inhibitor, increased ABCA1 protein but did not abolish linoleic acid-induced decrease in ABCA1 protein levels. Depletion of PKCδ using small interfering RNA showed decreased ABCA1 protein levels in control, palmitic acid-, and linoleic acid-treated cells; but the repressive effect of linoleic acid was sustained. In conclusion, our results indicate that unsaturated fatty acids regulate ABCA1 expression in HepG2 and FHs 74 Int cells at the posttranscriptional level and PKCδ is likely to be involved in maintaining ABCA1 protein levels.

Identification and characterization of small compound inhibitors of human FATP2.

Fatty acid transport proteins (FATPs) are bifunctional proteins, which transport long chain fatty acids into cells and activate very long chain fatty acids by esterification with coenzyme A. In an effort to understand the linkage between cellular fatty acid transport and the pathology associated with excessive accumulation of exogenous fatty acids, we targeted FATP-mediated fatty acid transport in a high throughput screen of more than 100,000 small diverse chemical compounds in yeast expressing human FATP2 (hsFATP2). Compounds were selected for their ability to depress the transport of the fluorescent long chain fatty acid analogue, C(1)-BODIPY-C(12). Among 234 hits identified in the primary screen, 5 compounds, each representative of a structural class, were further characterized in the human Caco-2 and HepG2 cell lines, each of which normally expresses FATP2, and in 3T3-L1 adipocytes, which do not. These compounds were effective in inhibiting uptake with IC(50)s in the low micromolar range in both Caco-2 and HepG2 cells. Inhibition of transport was highly specific for fatty acids and there were no effects of these compounds on cell viability, trans-epithelial electrical resistance, glucose transport, or long chain acyl-CoA synthetase activity. The compounds were less effective when tested in 3T3-L1 adipocytes suggesting selectivity of inhibition. These results suggest fatty acid transport can be inhibited in a FATP-specific manner without causing cellular toxicity.

Sitosterol reduces messenger RNA and protein expression levels of Niemann-Pick C1-like 1 in FHs 74 Int cells.

Intake of plant sterols has long been shown to reduce cholesterol absorption and subsequently plasma cholesterol concentrations. Despite competition between plant sterols and cholesterol for incorporation into mixed micelles as a suggested major mechanism for the inhibition of cholesterol absorption by plant sterols, studies exist to support an alternative mechanism. For example, another mechanism may be the action of plant sterols to reduce cholesterol absorption at the cellular level. This study was undertaken to test the hypothesis that plant sterols can modulate the expression of transporters such as Niemann-Pick C1-like 1 (NPC1L1) and scavenger receptor class B, type I (SR-BI) to lower intestinal cholesterol absorption. FHs 74 Int cells, a human small intestine epithelial cell line, were used as a model of enterocytes. The cells were treated with 25alpha-hydroxycholesterol (25 micromol/L) or 250 micromol/L of sitosterol, stigmasterol, and cholesterol for 24 hours to measure genes involved in cholesterol absorption and metabolism by quantitative real-time polymerase chain reaction. 25Alpha-hydroxycholesterol, cholesterol, and sitosterol significantly reduced the messenger RNA (mRNA) expression of NPC1L1 and hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, whereas SR-BI mRNA was not altered by the sterols. Western blot analysis confirmed the reduction in NPC1L1 by sterols. Depletion of cellular cholesterol by mevinolin, a cholesterol synthesis inhibitor, increased NPC1L1 and HMG-CoA reductase mRNA; and repletion of cholesterol abolished the increase. Sitosterol, but not stigmasterol, reduced the mRNA levels of NPC1L1 and HMG-CoA reductase to a similar extent of cholesterol. In conclusion, sitosterol can inhibit the expression of NPC1L1 in the enterocytes, which could be an alternate mechanism for plant sterols to reduce intestinal cholesterol uptake.

Endocannabinoids, metabolic regulation, and the role of diet.

Understanding the endocannabinoid system as it relates to health and disease is a relatively new area of study. The discovery and cloning of cannabinoid receptors have prompted an increase in research aimed at identifying endogenous ligands ("endocannabinoids") and how these receptors and ligands regulate a variety of physiologic and pathologic events that include bone formation, the cardiovascular system, appetite control, and energy metabolism. With regard to nutrition, researchers have begun to ask whether the known effects of diet on metabolic processes are mediated through endocannabinoids and their receptors. Although only a few studies have been conducted that directly address the role of diet, results indicate that endocannabinoids can be regulated by eating frequency and by specific dietary components, particularly fatty acids. This review provides an overview of the endocannabinoid system and its control of metabolism, with emphasis on the impact of diet.