Effect of quercetin on traits of the metabolic syndrome, endothelial function and inflammation in men with different APOE isoforms.

BACKGROUND AND AIMS: The polyphenol quercetin may prevent cardiovascular diseases due to its vasorelaxant and anti-oxidative properties. We investigated the effects of quercetin on risk factors of atherosclerosis, biomarkers of inflammation and oxidative stress, depending on the apolipoprotein E (APOE) genotype. METHODS AND RESULTS: In a double-blind crossover study 49 healthy male subjects with APOE genotype 3/3 (n = 19), 3/4 (n = 22) and 4/4 (n = 8) consumed 150 mg/d quercetin or placebo for 8 weeks each, intermitted by a three-week washout phase. After each intervention, endothelial function, anthropometry, metabolic and inflammatory parameters were measured in the fasting and postprandial state following a standardized lipid-rich meal. Endothelial function was not changed. In all subjects combined, quercetin significantly decreased waist circumference (P = 0.004) and postprandial systolic blood pressure (P = 0.044). Postprandial triacylglycerol concentrations were significantly decreased and HDL-cholesterol concentrations increased after quercetin as compared to placebo consumption (P = 0.025). Quercetin also moderately increased levels of TNFα (P = 0.024). There was a significant gene-diet interaction for waist circumference and for body mass index (BMI). CONCLUSIONS: Quercetin supplementation improved some risk factors of cardiovascular disease, yet exerted slightly pro-inflammatory effects. Genotype-dependent effects were seen only on waist circumference and BMI.

A promoter polymorphism in the liver-specific fatty acid transport protein 5 is associated with features of the metabolic syndrome and steatosis.

The fatty acid transport protein 5 (FATP5) is exclusively expressed in the liver and is involved in hepatic lipid and bile metabolism. We investigated whether a variation in the FATP5 promoter (rs56225452) is associated with hepatic steatosis and further features of the metabolic syndrome. A total of 716 male subjects from the Metabolic Intervention Cohort Kiel (MICK) and 103 male subjects with histologically proved nonalcoholic fatty liver disease (NAFLD) were genotyped for this FATP5 polymorphism rs56225452 and phenotyped for features of the metabolic syndrome. In the MICK cohort, ALT activities, postprandial insulin, and triglyceride concentrations were higher in subjects carrying the rare A-allele compared to GG homozygotes. Accordingly, the insulin sensitivity index determined after a mixed meal and standardized glucose load was lower in A-allele carriers. NAFLD cases carrying allele A were presented with also higher ALT activities. In NAFLD subjects, the association of BMI with the degree of steatosis and glucose concentration differed across FATP5 promoter polymorphism. The FATP5 promoter polymorphism rs56225452 is associated with higher ALT activity, insulin resistance, and dyslipidemia in the general population. The impact of the BMI on the severity of steatosis in NAFLD cases seems to depend on the FATP5 polymorphism.

Isomer-specific effects of CLA on gene expression in human adipose tissue depending on PPARgamma2 P12A polymorphism: a double blind, randomized, controlled cross-over study.

BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)gamma is a key regulator in adipose tissue. The rare variant Pro12Ala of PPARgamma2 is associated with a decreased risk of insulin resistance. Being dietary PPARgamma ligands, conjugated linoleic acids (CLAs) received considerable attention because of their effects on body composition, cancer, atherosclerosis, diabetes, obesity and inflammation, although some effects were only demonstrated in animal trials and the results in human studies were not always consistent. In the present study effects of CLA supplementation on genome wide gene expression in adipose tissue biopsies from 11 Ala12Ala and 23 Pro12Pro men were investigated. Subjects underwent four intervention periods (4 wk) in a randomized double blind cross-over design receiving 4.25 g/d of either cis-9, trans-11 CLA, trans-10,cis-12 CLA, 1:1 mixture of both isomers or a reference linoleic acid oil preparation. After each intervention biopsies were taken, whole genome expression microarrays were applied, and genes of interest were verified by realtime PCR. RESULTS: The following genes of lipid metabolism were regulated by CLA: LDLR, FASN, SCD, FADS1 and UCP2 were induced, while ABCA1, CD36 and CA3 were repressed. Transcription factors PPARgamma, NFAT5, CREB5 and EBF1, the adipokine NAMPT, members of the insulin signaling cascade SORBS1 and IGF1 and IL6ST were repressed, while the adipokine THBS1 and GLUT4 involved in insulin signaling were induced. Compared to trans-10,cis-12 CLA and the CLA mixture the cis-9, trans-11 CLA isomer exerted weaker effects. Only CD36 (-1.2 fold) and THBS1 (1.5 fold) were regulated. The CLA effect on expression of PPARgamma and leptin genes depends on the PPARgamma2 genotype. CONCLUSION: The data suggest that the isomer specific influence of CLA on glucose and lipid metabolism is genotype dependent and at least in part mediated by PPARgamma. TRIAL REGISTRATION: http://www.controlled-trials.com: ISRCTN91188075.

Milk and the metabolic syndrome.

The metabolic syndrome is a cluster of metabolic disorders, namely dyslipidaemia, hypertension, obesity and glucose intolerance. Insulin resistance is the core phenomenon. Co-occurrence is associated with increased cardiovascular disease (CVD) risk. Observational studies found no increased CVD risk with increasing consumption of milk and other dairy products. In several studies dairy consumption was inversely associated with the occurrence of one or several facets of the metabolic syndrome. Many dairy components may contribute to the beneficial effects. Milk and particularly whey appeared insulinotropic when given in a single meal, but not in longer-term intervention. Medium chain fatty acids improve insulin sensitivity. Whey proteins, amino acids, medium chain fatty acids and in particular calcium and other minerals may contribute to the beneficial effect of dairy products on body weight and body fat. Peptides, calcium and other minerals reduce blood pressure. Fermented products and probiotic bacteria decrease absorption of cholesterol, sphingomyelin of cholesterol and fat, calcium of cholesterol, bile acids and fat. Proteins, peptides and bacteria may also reduce plasma cholesterol. Lactose, citrate, proteins and peptides improve weight control, blood pressure and plasma lipids indirectly, by improving calcium bioavailability. Furthermore, dairy consumption improves the bioavailability of folate and other secondary plant components.

Determinants and impact of sleep duration in children and adolescents: data of the Kiel Obesity Prevention Study.

BACKGROUND/OBJECTIVES: This study investigates determinants of sleep duration and its impact on nutritional status, resting energy expenditure (REE), cardiometabolic risk factors and hormones in children/adolescents. SUBJECTS/METHODS: In 207 girls and 207 boys (13.0+/-3.4 (6.1-19.9) years) body mass index standard deviation score (BMI SDS), waist circumference (WC) z-score, body composition (air-displacement plethysmography), REE (ventilated hood system; n=312) and cardiometabolic risk factors/hormones (n=250) were assessed. Greater than 90th percentile of BMI/WC references was defined as overweight/overwaist. Sleep duration, media consumption (TV watching/computer use), physical activity, dietary habits, parental BMI, socio-economic status and early infancy were assessed by questionnaire. Short sleep was defined as <10 h per day for children <10 years and otherwise <9 h per day. RESULTS: Total 15.9% participants were overweight, mean sleep duration was 8.9+/-1.3 h per day. Age explained most variance in sleep (girls: 57.0%; boys: 41.2%) besides a high nutrition quality score (girls: 0.9%) and a low media consumption (boys: 1.3%). Sleep was inversely associated with BMI SDS/WC z-score (girls: r=-0.17/-0.19, P<0.05; boys: r=-0.21/-0.20, P<0.01), which was strengthened after adjusting for confounders. Short vs long sleep was associated with 5.5-/2.3-fold higher risks for obesity/overwaist (girls). After adjusting for age, REE (adjusted for fat-free mass) was positively associated with sleep in boys (r=0.16, P<0.05). Independently of age and WC z-score, short sleep was associated with lower adiponectin levels in boys (11.7 vs 14.4 microg/ml, P<0.05); leptin levels were inversely related to sleep in girls (r=-0.23, P<0.05). Homoeostasis model assessment-insulin resistance (r=-0.20, P<0.05) and insulin levels (r=-0.20, P<0.05) were associated with sleep (girls), which depended on WC z-score. CONCLUSIONS: Age mostly determined sleep. Short sleep was related to a higher BMI SDS/WC z-score (girls/boys), a lower REE (boys), higher leptin (girls) and lower adiponectin levels (boys).

Common familial influences on clustering of metabolic syndrome traits with central obesity and insulin resistance: the Kiel obesity prevention study.

OBJECTIVE: The phenotypic heterogeneity of metabolic syndrome (MSX) suggests heterogeneity of the underlying genotype. The aim of the present study was to examine the common genetic background that contributes to the clustering between the two main features (insulin resistance, central obesity) and different MSX component traits. METHODS: In all, 492 individuals from 90 families were investigated in a three-generation family path study as part of the Kiel Obesity Prevention Study (KOPS, 162 grandparents, 66.1+/-6.7 years, 173 parents, 41.3+/-5.4 years and 157 children, 10.8+/-3.4 years). Overall heritability was estimated and common familial (genetic and environmental) influences on insulin resistance (HOMA-IR) or central obesity (elevated waist circumference, WC), respectively, and different MSX traits were compared in a bivariate cross-trait correlation model. RESULTS: Prevalence of MSX (according to NCEP criteria) was 27.2% (f) and 27.8% (m) in adults and 3.5% (f) and 8.5% (m) in children and adolescents, respectively. MSX phenotype was found to be highly variable, comprising 16 subtypes of component trait combinations. Within-trait heritability was 38.5% for HOMA-IR and 53.5% for WC, cross-trait heritability was 53.4%. As much as 6-18% and 3-10% of the shared variance between different MSX component traits (lipid profile, blood pressure) and WC or HOMA-IR, respectively, may be genetic. With the exception of HDL-C, the shared genetic variance between MSX component traits and WC was higher than the genetic variance shared with HOMA-IR. CONCLUSION: A common genetic background contributes to the clustering of different MSX component traits and central obesity or insulin resistance. Common genetic influences favour central obesity as a major characteristic linking these traits.

Bioactive substances in milk with properties decreasing risk of cardiovascular diseases.

Milk is often seen as a potential promotor of atherosclerosis and coronary heart disease because it is a source of cholesterol and saturated fatty acids. But there are several studies indicating that milk and milk products may not affect adversely blood lipids as would be predicted from its fat content and fat composition. There are even factors in milk and milk products which may actively protect from this condition by improving several risk factors. Calcium, bioactive peptides and as yet unidentified components in whole milk may protect from hypertension, and folic acid, vitamin B6 (pyridoxine) and B12 (cyanocobalamin) or other unidentified components of skim milk may contribute to low homocysteine levels. Conjugated linoleic acid may have hypolipidaemic and antioxidative and thus antiatherosclerotic properties. Epidemiological studies suggest that milk and milk products fit well into a healthy eating pattern emphasizing cereals and vegetables.

Dietary protein and atherogenesis.

Different dietary proteins determine different serum cholesterol levels if fed in a semisynthetic diet to some, but not all, animal species. In one species, the rabbit, this metabolic response is elicited without adding high sucrose or cholesterol supplements that have to be added to rat or pig diets in order to cause a similar response. Eleven out of 13 studies show that casein and soy protein do not induce different serum cholesterol levels in normal man. More important, protein-induced differences of serum cholesterol concentrations have not been reported when appropriate nutritional methodology has been applied. We conclude that no protein-induced hypercholesterolemia is observed in primates, particularly not in the human species. Dietary recommendations urging the general public to reduce consumption of animal protein because of a higher atherogenicity are not supported by the present data. The biochemical basis of the metabolic responses has been studied by many investigators, but no convincing unifying concept has yet been identified. The recent observation of higher serum thyroxine concentrations following soy protein consumption (and vegetable protein in general) when compared to casein shed new light on this problem. This endocrine response explains a wide array of metabolic features of soy-fed rodents: the lower hepatic VLDL secretion, the higher hepatic HMG-CoA reductase activity, the higher hepatic apo B, E receptor activity, the higher fecal bile acid excretion, and finally the lower serum cholesterol concentrations.

Labeling with 15N as compared with homoarginine suggests a lower prececal digestibility of casein in pigs.

Calculations of prececal protein digestibility based on the stable isotope 15N and the chemical label homoarginine were compared, using casein doubly-labeled with both markers. After food was withheld overnight 24 miniature pigs were given a meal containing 15 g/100 g casein, including 4 g/100 g doubly-labeled protein, and chromic oxide as an indigestible marker. The intestine of eight animals each was removed 3, 6 or 12 h later, divided into 3 sections of equal length, and chyme was collected. Kjeldahl-N, 15N and homoarginine were determined in diet and chyme. Digestibility of casein in the distal third of the small intestine was 93.5 +/- 0.5% and 97.6 +/- 0.3% (P < 0.05) according to 15N and homoarginine label, respectively. Potential causes for this systematic difference were assessed. The data suggest that incorporation of 15N into endogenous proteins and re-entry into the intestinal lumen via secreta and desquamations is the major cause for the 4.2 +/- 0.4% lower digestibility based on the 15N as compared with the homoarginine labeling technique. A preferential occurrence of homoarginine in more easily digestible sections of the protein, faster release during the digestive process and absorption of homoarginine, or incorporation of 15N into proteins of intestinal bacteria are less likely to cause this difference.

Modulation of very low-density lipoprotein secretion by dietary protein is age-dependent in rats.

The Triton WR-1339 technique was used in order to study age-dependent changes of the rate of very low-density lipoprotein (VLDL) lipid secretion in rats consuming either casein or soy protein isolate. There was a significantly higher influx of lipoprotein cholesterol and triglycerides into the plasma compartment (of fasted animals) after a casein than after a soy diet, both in 10-week-old animals (cholesterol: 0.78 +/- 0.06 vs. 0.52 +/- 0.03 mmol X 1(-1) X h-1) and 25-week-old animals (cholesterol: 0.50 +/- 0.07 vs. 0.32 +/- 0.05 mmol X 1(-1) X h-1), but secretion rates diminished with age. The higher secretion rates following casein were paralleled by higher serum cholesterol levels in the fasted younger animals and also, though to a lesser degree, in the fed animals. These data contribute further evidence to our earlier proposal that dietary protein may induce different serum lipid levels by the modulating rate of lipid influx into the plasma compartment.

Dietary sucrose but not starch promotes protein-induced differences in rates of VLDL secretion and plasma lipid concentrations in rats.

Casein instead of soybean protein in a low fat, cholesterol-free diet containing sucrose increases both lipid secretion by rat liver and plasma cholesterol concentration. Male rats were studied to determine whether dietary sucrose is a prerequisite for these protein-induced differences of lipid metabolism in vivo. After 6 wk of consuming either casein plus sucrose, soybean protein isolate plus sucrose, casein plus starch or soybean protein isolate plus starch (carbohydrate, 69 g/100 g diet; protein, 20 g/100 g diet) plasma lipids were measured. VLDL degradation was blocked by Triton WR-1339 and rates of lipid secretion were calculated from the observed linear increase in plasma levels with time. Plasma cholesterol concentrations and rates of secretion of VLDL cholesterol and triglycerides were 27, 47 and 34% higher, respectively, in animals fed casein plus sucrose compared with soybean protein plus sucrose. These differences were clearly diminished or absent when starch was the dietary carbohydrate. The data substantiate the finding that dietary sucrose but not dietary starch promotes a casein-induced increase in hepatic cholesterol secretion and thus contributes to casein-induced hypercholesterolemia in rats. On the contrary, differences of triglyceride secretion were not reflected in different plasma lipid concentrations.

The hypocholesterolemic effect of guar gum depends on dietary sucrose--studies in minipigs.

We examined the effect of guar gum on serum lipids if fed together with either 50.3% starch or 50.3% sucrose in a balanced diet to pigs. For this purpose, five adult hypercholesterolemic minipigs (total serum cholesterol 9.0 mmol/l) underwent three consecutive 8-week crossover (control or guar supplementation) feeding experiments (experiment I = cornstarch plus 15 g guar, experiment II = cornstarch plus 30 g guar, experiment III = sucrose plus 30 g guar per day). With the cornstarch-based diet neither 15 g nor 30 g guar gum had an influence on serum total cholesterol or triglyceride levels. Also, the cholesterol concentrations in the lipoprotein fractions did not change significantly during experiments I and II, yet total serum cholesterol concentration was about 20% lower (p less than 0.01) when guar gum was added to the sucrose diet in experiment III. In the presence of sucrose the supplementation of 30 g guar led to a significant decrease (p less than 0.05) of the cholesterol concentrations in the very low-density lipoproteins (VLDL) and high-density lipoproteins (HDL). There was also a tendency for decreased cholesterol levels in the low-density lipoproteins (LDL) after adding 30 g guar to the sucrose diet. Thus, the study demonstrates that guar gum exerts a hypocholesterolemic effect in the presence of sucrose in the diet, but not in the case of starch consumption.

The effect of a free mackerel-supplemented diet on plasma and lipoprotein lipid concentrations in normolipidemic subjects.

The effect of mackerel consumption on plasma and lipoprotein lipid concentrations was studied in a seven-day experiment in eight healthy, normolipidemic subjects. Participants ate about 100 g mackerel (corresponding to about 2.5-3 g omega 3 fatty acids daily. The mean triglyceride concentrations in total plasma, VLDL, and LDL were significantly reduced by 40, 46.7, and 38.5% respectively after fish consumption. There was also a small (non-significant) reduction of plasma cholesterol and a (significant) increase of the ratio of HDL/total plasma cholesterol. These data show that a moderately increased intake of omega-3 fatty acids by fish food can change lipid characteristics in healthy normolipidemic individuals within a short-time period, even on a free diet.

Influence of different dietary proteins on plasma growth hormone in rats.

Dietary casein, compared to vegetable protein, causes hypercholesterolemia in some animal species. This may be associated with a change of hormonal status. Among others, GH has an important impact on cholesterol metabolism; GH deficiency results in hypercholesterolemia. This paper shows that the rhythmic variation of GH levels in rats is differently affected by different dietary proteins. Within a 4-h observation period overall mean values and integrated areas under the GH levels plotted against time are higher with casein as compared to soy protein. Secretory GH peak values are lower than reported before for chow-fed rats. These observations support the idea that different dietary proteins cause a different endocrine response. As GH levels are higher with casein, while lower levels would be expected to be associated with hypercholesterolemia, the observed differences are obviously of less relevance for the expression of casein-induced hypercholesterolemia.

Influence of casein versus soy protein isolate on lipid metabolism of minipigs.

This report describes whether a complete exchange of soy protein isolate for casein affects serum lipid or lipoprotein levels. For this purpose, 10 adult minipigs underwent two 6-week crossover experiments, one with 11 and one with 22 weight % protein in a western-style diet. Cholesterol levels were 2.02 +/- 0.06 and 1.98 +/- 0.04 mmol/l with casein and soy, respectively (22 weight %) and 2.08 +/- 0.06 and 2.04 +/- 0.05 mmol/l, respectively (11 weight %). There was a significant rise of high-density lipoprotein cholesterol by the lower protein intake, both with casein and soy. In all experiments nitrogen balance was positive. Soy feeding was accompanied by a significant rise of fecal bacterial protein excretion, as calculated from diaminopimelic acid. In summary, no statistically significant difference of plasma or low-density lipoprotein cholesterol levels were observed, neither due to the kind nor to the amount of dietary protein.

Effects of oral and intracecal pectin administration on blood lipids in minipigs.

The effects on blood lipids of oral and intracecal pectin administration for 4 wk were examined in six male adult hypercholesterolemic minipigs (total serum cholesterol = 8.4 mmol/L. The pigs were fitted with a cecal cannula and were assigned to three groups of two pigs each; the groups underwent an experiment in 3 X 3 Latin square design. The control period was designated as that during which water was infused into the cecum. Total serum cholesterol concentration was about 50% lower after feeding 75 g pectin per day than during the control period. The intracecal infusion of the same amount of pectin had no effect on total serum cholesterol concentration compared to the control period. The cholesterol level in the low density lipoproteins (LDL), in the very low density lipoproteins (VLDL) and in the high density lipoproteins (HDL) was lower after feeding pectin than in the control period. The HDL cholesterol, but not LDL or VLDL cholesterol, was at a lower level after intracecal infusion of pectin than during the control period. Thus, the study clearly demonstrates that the passage of pectin through the small intestine is necessary for its hypocholesterolemic action.

Transport of HDL cholesterol esters to the liver is not diminished by probucol treatment in rats.

This study examined the relation of decreased high density lipoprotein (HDL) levels in probucol-fed rats and the transport of HDL cholesterol esters (CEs) to the liver. HDLs from both control rats and rats fed 1% probucol for 3 weeks were doubly labeled in their CE and apolipoprotein A-I moieties with intracellularly trapped tracers and then intravenously injected into probucol-fed or control rats for determination of plasma decay kinetics and sites of tracer uptake. Results for HDL from control and probucol-fed rats were not different. The fractional catabolic rate (FCR) of plasma HDL CE was significantly increased by probucol feeding (23%) so that mass transport of HDL CE through the plasma compartment was not significantly different from that in control rats. The plasma FCR for apolipoprotein A-I did not change. Similarly, the FCR for uptake of HDL CE by the liver increased on probucol feeding (20%), resulting in a near-normal rate of HDL CE mass uptake, whereas the FCR for HDL particle uptake (measured by apolipoprotein A-I uptake) did not change. Thus, the maintenance of near-normal HDL CE uptake by the liver was exclusively due to increased selective uptake (32%). To the extent that hepatic uptake of HDL CE mediates reverse cholesterol transport, that process was not significantly compromised in rats fed 1% probucol.

Probucol increases the selective uptake of HDL cholesterol esters by Hep G2 human hepatoma cells.

A previous study in rats showed that even though probucol substantially lowers high density lipoprotein (HDL) levels, near-normal mass transport of HDL cholesterol esters (CE) to the liver is maintained by the induction of "selective" (direct) uptake of HDL CE. The present study describes a parallel result in cultured Hep G2 human hepatoma cells. Cells were preincubated in the presence or absence of probucol before measuring the uptake of doubly labeled HDL3 in the absence of probucol. Preincubation with probucol decreased the uptake of HDL3 particles (iodine-125-labeled N-methyltyramine cellobiose-apolipoprotein [125I-NMTC-apo] A-I uptake) but increased the uptake of [3H]cholesteryl oleyl ether in excess of 125I-NMTC-apo A-I (i.e., selective uptake) in a dose-dependent fashion. The reversibly cell-associated pool of CE tracer, a precursor for selective uptake, enlarged on probucol treatment, but the increase was not in proportion to the increase in selective uptake. HDL3 particle uptake decreased on probucol treatment. The decrease was evident after less than 20 minutes of probucol exposure and was maximal after 6 hours; in contrast, HDL3 CE selective uptake increased only after greater than 13 hours and had not reached a plateau after 20 hours. Thus, effects on particle uptake and selective uptake were dissociated in time.

Influence of dietary casein or soy protein on serum lipids and lipoproteins of monkeys (Macaca fascicularis).

In order to learn whether a complete exchange of dietary soy protein for casein causes a change of serum lipid levels in primates, 8 Macaca fascicularis monkeys were fed a semi-purified ration; its basic composition was similar to diets consumed in Western Europe. In a crossover experiment, casein was exchanged for a soy protein isolate and vice versa. Plasma and lipoprotein cholesterol and triglyceride levels were determined following a 3-week dietary regimen. There was no significant difference of total serum cholesterol levels between the soy protein and casein periods (4.6 vs. 4.7 mmol/l). However, high-density lipoprotein (HDL) cholesterol levels were significantly higher (1.6 vs. 1.2 mmol/l, p less than 0.01) and very-low-density lipoprotein (VLDL) cholesterol levels were significantly lower (0.39 vs. 0.55 mmol/l, p less than 0.01) following the soy protein diet. There was no distinct change in total or lipoprotein triglyceride levels following exchange of the dietary protein. It is concluded that in nonhuman primates a complete exchange of soy protein for casein produces changes of HDL, but not of total or low-density lipoprotein (LDL) cholesterol.