Peripheral blood mononuclear cells as a source to detect markers of homeostatic alterations caused by the intake of diets with an unbalanced macronutrient composition
Diaz-Rua, R. ; Keijer, J. ; Caimari, A. ; Schothorst, E.M. van; Oliver, P. ; Palou, A. - \ 2015
Journal of Nutritional Biochemistry 26 (2015)4. - ISSN 0955-2863 - p. 398 - 407.
high-fat-diet - high-protein diet - gene-expression profiles - preadipocyte factor-i - beta-casein 1-28 - insulin-resistance - oxidative stress - adipose-tissue - antigen presentation - energy homeostasis
Peripheral blood mononuclear cells (PBMC) are accessible in humans and their gene expression pattern was shown to reflect overall physiological response of the body to a specific stimulus, such as diet. We aimed to study the impact of sustained intake (4 months) of diets with an unbalanced macronutrient proportion (rich in fat or protein) administered isocalorically to a balanced control diet, as physiological stressors on PBMC whole genome gene expression in rats, to better understand the effects of these diets on metabolism and health and to identify biomarkers of nutritional imbalance. Dietary macronutrient composition (mainly increased protein content) altered PBMC gene expression, with genes involved in immune response being the most affected. Intake of a high-fat (HF) diet decreased the expression of genes related to antigen recognition/presentation, whereas the high-protein (HP) diet increased the expression of these genes and of genes involved in cytokine signaling and immune system maturation/activation. Key energy homeostasis genes (mainly related to lipid metabolism) were also affected, reflecting an adaptive response to the diets. Moreover, HF diet feeding impaired expression of genes involved in redox balance regulation. Finally, we identified a common gene expression signature of 7 genes whose expression changed in the same direction in response to the intake of both diets. These genes, individually or together, constitute a potential risk marker of diet macronutrient imbalance. In conclusion, we newly show that gene expression analysis in PBMC allows detection of diet-induced physiological deviations that distinguish from a diet with a proper and equilibrated macronutrient composition.
A weekly alternating diet between caloric restriction and medium-fat protects the liver from fatty liver development in middle-aged C57BL/6J mice
Rusli, F. ; Boekschoten, M.V. ; Zubia, A.A. ; Lute, C. ; Müller, M.R. ; Steegenga, W.T. - \ 2015
Molecular Nutrition & Food Research 59 (2015)3. - ISSN 1613-4125 - p. 533 - 543.
metabolic syndrome - insulin-resistance - small-intestine - induced obesity - adipose-tissue - life-span - disease - prevalence - population - expression
Scope : We aimed to investigate whether a novel dietary intervention consisting of an every-other-week calorie restricted diet could prevent non-alcoholic fatty liver disease (NAFLD) development induced by a medium-fat diet. Methods and results : Nine week-old male C57BL/6J mice received either a 1) control (C), 2) 30E% calorie restricted (CR), 3) medium-fat (MF; 25E% fat) or 4) intermittent (INT) diet, a diet alternating weekly between 40E% CR and an ad libitum MF diet until sacrifice at the age of 12 months. The metabolic, morphological, and molecular features of NAFLD were examined. The INT diet resulted in healthy metabolic and morphological features as displayed by the continuous CR diet: glucose tolerant, low hepatic triglyceride content, low plasma alanine aminotransferase. In contrast, the C- and MF-exposed mice with high body weight developed signs of NAFLD. However, the gene expression profiles of INT-exposed mice differed to those of CR-exposed mice and showed to be more similar with those of C- and MF-exposed mice with a comparable body weight. Conclusions : Our study reveals that the INT diet maintains metabolic health and reverses the adverse effects of the MF diet, thus effectively prevent the development of NAFLD in 12-month-old male C57BL/6J mice.
Oxygen restriction as challenge test reveals early high-fat-diet-induced changes in glucose and lipid metabolism
Duivenvoorde, L.P.M. ; Schothorst, E.M. van; Derous, D. ; Stelt, I. van der; Masania, J. ; Rabbani, N. ; Thornalley, P.J. ; Keijer, J. - \ 2015
Pflugers Archiv-European Journal of Physiology 467 (2015)6. - ISSN 0031-6768 - p. 1179 - 1193.
adipose-tissue - gene-expression - intermittent hypoxia - energy-expenditure - insulin-resistance - transcriptional regulation - mass-spectrometry - mice - obesity - oxidation
Challenge tests stress homeostasis and may reveal deviations in health that remain masked under unchallenged conditions. Ideally, challenge tests are non-invasive and applicable in an early phase of an animal experiment. Oxygen restriction (OxR; based on ambient, mild normobaric hypoxia) is a non-invasive challenge test that measures the flexibility to adapt metabolism. Metabolic inflexibility is one of the hallmarks of the metabolic syndrome. To test whether OxR can be used to reveal early diet-induced health effects, we exposed mice to a low-fat (LF) or high-fat (HF) diet for only 5 days. The response to OxR was assessed by calorimetric measurements, followed by analysis of gene expression in liver and epididymal white adipose tissue (eWAT) and serum markers for e.g. protein glycation and oxidation. Although HF feeding increased body weight, HF and LF mice did not differ in indirect calorimetric values under normoxic conditions and in a fasting state. Exposure to OxR; however, increased oxygen consumption and lipid oxidation in HF mice versus LF mice. Furthermore, OxR induced gluconeogenesis and an antioxidant response in the liver of HF mice, whereas it induced de novo lipogenesis and an antioxidant response in eWAT of LF mice, indicating that HF and LF mice differed in their adaptation to OxR. OxR also increased serum markers of protein glycation and oxidation in HF mice, whereas these changes were absent in LF mice. Cumulatively, OxR is a promising new method to test food products on potential beneficial effects for human health.
Assessment of Metabolic Flexibility of Old and Adult Mice Using Three Noninvasive, Indirect Calorimetry-Based Treatments
Duivenvoorde, L.P.M. ; Schothorst, E.M. van; Swarts, J.J.M. ; Keijer, J. - \ 2015
Journals of Gerontology. Series A: Biological Sciences & Medical Sciences 70 (2015)3. - ISSN 1079-5006 - p. 282 - 293.
type-2 diabetes-mellitus - adipose-tissue - energy-metabolism - gene-expression - dietary restriction - hypobaric hypoxia - laboratory mice - down-regulation - weight-gain - food-intake
Indirect calorimetry (InCa) can potentially be used to noninvasively assess metabolic and age-related flexibility. To assess the use of InCa for this purpose, we tested the sensitivity and response stability over time of three InCa-based treatments in old versus adult mice. Diurnal patterns of respiratory exchange ratio were followed for 24 hours under standard conditions (Treatment 1), but the results were not stable between test periods. As a challenge, fasted mice received glucose to test switch-effectiveness from fat to glucose oxidation (Treatment 2). No differences between groups were observed, although old mice showed higher adiposity and lower white adipose tissue (WAT) mitochondrial density, indicative of age-impaired metabolic health. Lastly, adaptation to a challenge of oxygen restriction (OxR, 14.5% O2) was assessed as a novel approach (Treatment 3). This treatment stably detected significant differences: old mice did not maintain reduced oxygen consumption under OxR during both test periods, whereas adult mice did. Further biochemical and gene expression analyses showed that OxR affected glucose and lactate homeostasis in liver and WAT of adult mice, supporting the observed differences in oxygen consumption. In conclusion, InCa analysis of the response to OxR in mice is a sensitive and reproducible treatment to noninvasively measure age-impaired metabolic health.
Circulating angiopoietin-like 4 links proteinuria with hypertriglyceridemia in nephrotic syndrome
Clement, L.C. ; Mace, C. ; Avila-Casado, C. ; Joles, J.A. ; Kersten, A.H. ; Chugh, S.S. - \ 2014
Nature Medicine 20 (2014)1. - ISSN 1078-8956 - p. 37 - 46.
lipoprotein-lipase - fatty-acids - gene-expression - adipose-tissue - angptl4 - disease - serum - rats - mice - glomeruli
The molecular link between proteinuria and hyperlipidemia in nephrotic syndrome is not known. We show in the present study that plasma angiopoietin-like 4 (Angptl4) links proteinuria with hypertriglyceridemia through two negative feedback loops. In previous studies in a rat model that mimics human minimal change disease, we observed localized secretion by podocytes of hyposialylated Angptl4, a pro-proteinuric form of the protein. But in this study we noted high serum levels of Angptl4 (presumably normosialylated based on a neutral isoelectric point) in other glomerular diseases as well. Circulating Angptl4 was secreted by extrarenal organs in response to an elevated plasma ratio of free fatty acids (FFAs) to albumin when proteinuria reached nephrotic range. In a systemic feedback loop, these circulating pools of Angptl4 reduced proteinuria by interacting with glomerular endothelial alpha(v)beta(5) integrin. Blocking the Angptl4-beta(5) integrin interaction or global knockout of Angptl4 or beta(5) integrin delayed recovery from peak proteinuria in animal models. But at the same time, in a local feedback loop, the elevated extrarenal pools of Angptl4 reduced tissue FFA uptake in skeletal muscle, heart and adipose tissue, subsequently resulting in hypertriglyceridemia, by inhibiting lipoprotein lipase (LPL)-mediated hydrolysis of plasma triglycerides to FFAs. Injecting recombinant human ANGPTL4 modified at a key LPL interacting site into nephrotic Buffalo Mna and Zucker Diabetic Fatty rats reduced proteinuria through the systemic loop but, by bypassing the local loop, without increasing plasma triglyceride levels. These data show that increases in circulating Angptl4 in response to nephrotic-range proteinuria reduces the degree of this pathology, but at the cost of inducing hypertriglyceridemia, while also suggesting a possible therapy to treat these linked pathologies.
Biomarker patterns of inflammatory and metabolic pathways are associated with risk of colorectal cancer: results from the European Prospective Investigation into Cancer and Nutrition (EPIC)
Aleksandrova, K. ; Jenab, M. ; Bueno-de-Mesquita, H.B. ; Fedirko, V. ; Kaaks, R. ; Lukanova, A. ; Duijnhoven, F.J.B. van - \ 2014
European Journal of Epidemiology 29 (2014)4. - ISSN 0393-2990 - p. 261 - 275.
soluble leptin receptor - density-lipoprotein cholesterol - c-reactive protein - insulin-resistance - oxidative stress - hdl-cholesterol - multiple imputation - plasma adiponectin - binding-proteins - adipose-tissue
A number of biomarkers of inflammatory and metabolic pathways are individually related to higher risk of colorectal cancer (CRC); however, the association between biomarker patterns and CRC incidence has not been previously evaluated. Our study investigates the association of biomarker patterns with CRC in a prospective nested case-control study within the European Prospective Investigation into Cancer and Nutrition (EPIC). During median follow-up time of 7.0 (3.7-9.4) years, 1,260 incident CRC cases occurred and were matched to 1,260 controls using risk-set sampling. Pre-diagnostic measurements of C-peptide, glycated hemoglobin, triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), C-reactive protein (CRP), reactive oxygen metabolites (ROM), insulin-like growth factor 1, adiponectin, leptin and soluble leptin receptor (sOB-R) were used to derive biomarker patterns from principal component analysis (PCA). The relation with CRC incidence was assessed using conditional logistic regression models. We identified four biomarker patterns 'HDL-C/Adiponectin fractions', 'ROM/CRP', 'TG/C-peptide' and 'leptin/sOB-R' to explain 60 % of the overall biomarker variance. In multivariable-adjusted logistic regression, the 'HDL-C/Adiponectin fractions', 'ROM/CRP' and 'leptin/sOB-R' patterns were associated with CRC risk [for the highest quartile vs the lowest, incidence rate ratio (IRR) = 0.69, 95 % CI 0.51-0.93, P-trend = 0.01; IRR = 1.70, 95 % CI 1.30-2.23, P-trend = 0.002; and IRR = 0.79, 95 % CI 0.58-1.07; P-trend = 0.05, respectively]. In contrast, the 'TG/C-peptide' pattern was not associated with CRC risk (IRR = 0.75, 95 % CI 0.56-1.00, P-trend = 0.24). After cases within the first 2 follow-up years were excluded, the 'ROM/CRP' pattern was no longer associated with CRC risk, suggesting potential influence of preclinical disease on these associations. By application of PCA, the study identified 'HDL-C/Adiponectin fractions', 'ROM/CRP' and 'leptin/sOB-R' as biomarker patterns representing potentially important pathways for CRC development.
Nutritional aspects of metabolic inflammation in relation to health-insights from transcriptomic biomarkers in PBMC of fatty acids and polyphenols
Afman, L.A. ; Milenkovic, D. ; Roche, H. - \ 2014
Molecular Nutrition & Food Research 58 (2014)8. - ISSN 1613-4125 - p. 1708 - 1720.
blood mononuclear-cells - gene-expression profiles - fish-oil supplementation - improves insulin sensitivity - randomized controlled-trial - coronary-artery-disease - adipose-tissue - postmenopausal women - nlrp3 inflammasome - in-vivo
Recent research has highlighted potential important interaction between metabolism and inflammation, within the context of metabolic health and nutrition, with a view to preventing diet-related disease. In addition to this, there is a paucity of evidence in relation to accurate biomarkers that are capable of reflecting this important biological interplay or relationship between metabolism and inflammation, particularly in relation to diet and health. Therefore the objective of this review is to highlight the potential role of transcriptomic approaches as a tool to capture the mechanistic basis of metabolic inflammation. Within this context, this review has focused on the potential of peripheral blood mononuclear cells transcriptomic biomarkers, because they are an accessible tissue that may reflect metabolism and subacute chronic inflammation. Also these pathways are often dysregulated in the common diet-related diseases obesity, type 2 diabetes, and cardiovascular disease, thus may be used as markers of systemic health. The review focuses on fatty acids and polyphenols, two classes of nutrients/nonnutrient food components that modulate metabolism/inflammation, which we have used as an example of a proof-of-concept with a view to understanding the extent to which transcriptomic biomarkers are related to nutritional status and/or sensitive to dietary interventions. We show that both nutritional components modulate inflammatory markers at the transcriptomic level with the capability of profiling pro- and anti-inflammatory mechanisms in a bidirectional fashion; to this end transcriptomic biomarkers may have potential within the context of metabolic inflammation. This transcriptomic biomarker approach may be a sensitive indicator of nutritional status and metabolic health.
Adipocyte spliced form of X-box-binding protein 1 promotes adiponectin multimerization and systemic glucose homeostasis
Sha, H. ; Yang, L. ; Liu, M. ; Xia, S. ; Liu, Y. ; Liu, F. ; Kersten, A.H. ; Qi, L. - \ 2014
Diabetes 63 (2014)3. - ISSN 0012-1797 - p. 867 - 879.
endoplasmic-reticulum stress - plasma-cell differentiation - transcription factor xbp-1 - links er stress - down-regulation - messenger-rna - dsba-l - insulin sensitivity - adipose-tissue - obesity
The physiological role of the spliced form of X-box–binding protein 1 (XBP1s), a key transcription factor of the endoplasmic reticulum (ER) stress response, in adipose tissue remains largely unknown. In this study, we show that overexpression of XBP1s promotes adiponectin multimerization in adipocytes, thereby regulating systemic glucose homeostasis. Ectopic expression of XBP1s in adipocytes improves glucose tolerance and insulin sensitivity in both lean and obese (ob/ob) mice. The beneficial effect of adipocyte XBP1s on glucose homeostasis is associated with elevated serum levels of high-molecular-weight adiponectin and, indeed, is adiponectin-dependent. Mechanistically, XBP1s promotes adiponectin multimerization rather than activating its transcription, likely through a direct regulation of the expression of several ER chaperones involved in adiponectin maturation, including glucose-regulated protein 78 kDa, protein disulfide isomerase family A, member 6, ER protein 44, and disulfide bond oxidoreductase A–like protein. Thus, we conclude that XBP1s is an important regulator of adiponectin multimerization, which may lead to a new therapeutic approach for the treatment of type 2 diabetes and hypoadiponectinemia.
Nutrigenomics of Body Weight Regulation: A Rationale for Careful Dissection of Individual Contributors
Keijer, J. ; Hoevenaars, F.P.M. ; Nieuwenhuizen, A.G. ; Schothorst, E.M. van - \ 2014
Nutrients 6 (2014)10. - ISSN 2072-6643 - p. 4531 - 4551.
blood mononuclear-cells - diet-induced obesity - high-fat diet - adipose-tissue - metabolic-rate - adaptive thermogenesis - food-intake - nutrition transition - energy-requirements - mass-spectrometry
Body weight stability may imply active regulation towards a certain physiological condition, a body weight setpoint. This interpretation is ill at odds with the world-wide increase in overweight and obesity. Until now, a body weight setpoint has remained elusive and the setpoint theory did not provide practical clues for body weight reduction interventions. For this an alternative theoretical model is necessary, which is available as the settling point model. The settling point model postulates that there is little active regulation towards a predefined body weight, but that body weight settles based on the resultant of a number of contributors, represented by the individual’s genetic predisposition, in interaction with environmental and socioeconomic factors, such as diet and lifestyle. This review refines the settling point model and argues that by taking body weight regulation from a settling point perspective, the road will be opened to careful dissection of the various contributors to establishment of body weight and its regulation. This is both necessary and useful. Nutrigenomic technologies may help to delineate contributors to body weight settling. Understanding how and to which extent the different contributors influence body weight will allow the design of weight loss and weight maintenance interventions, which hopefully are more successful than those that are currently available.
The COLON study: Colorectal cancer: Longitudinal, Observational study on Nutritional and lifestyle factors that may influence colorectal tumour recurrence, survival and quality of life
Winkels, R.M. ; Heine-Bröring, R.C. ; Zutphen, M. van; Harten-Gerritsen, A.S. van; Kok, D.E.G. ; Duijnhoven, F.J.B. van; Kampman, E. - \ 2014
BMC Cancer 14 (2014). - ISSN 1471-2407 - 8 p.
body-mass index - enhancing physical-activity - treatment-related toxicity - visceral obesity - short questionnaire - skeletal-muscle - clinical-trials - adipose-tissue - rectal-cancer - health
Background There is clear evidence that nutrition and lifestyle can modify colorectal cancer risk. However, it is not clear if those factors can affect colorectal cancer treatment, recurrence, survival and quality of life. This paper describes the background and design of the “COlorectal cancer: Longitudinal, Observational study on Nutritional and lifestyle factors that may influence colorectal tumour recurrence, survival and quality of life” – COLON – study. The main aim of this study is to assess associations of diet and other lifestyle factors, with colorectal cancer recurrence, survival and quality of life. We extensively investigate diet and lifestyle of colorectal cancer patients at diagnosis and during the following years; this design paper focusses on the initial exposures of interest: diet and dietary supplement use, body composition, nutrient status (e.g. vitamin D), and composition of the gut microbiota. Methods/Design The COLON study is a multi-centre prospective cohort study among at least 1,000 incident colorectal cancer patients recruited from 11 hospitals in the Netherlands. Patients with colorectal cancer are invited upon diagnosis. Upon recruitment, after 6 months, 2 years and 5 years, patients fill out food-frequency questionnaires; questionnaires about dietary supplement use, physical activity, weight, height, and quality of life; and donate blood samples. Diagnostic CT-scans are collected to assess cross-sectional areas of skeletal muscle, subcutaneous fat, visceral fat and intermuscular fat, and to assess muscle attenuation. Blood samples are biobanked to facilitate future analyse of biomarkers, nutrients, DNA etc. Analysis of serum 25-hydroxy vitamin D levels, and analysis of metabolomic profiles are scheduled. A subgroup of patients with colon cancer is asked to provide faecal samples before and at several time points after colon resection to study changes in gut microbiota during treatment. For all patients, information on vital status is retrieved by linkage with national registries. Information on clinical characteristics is gathered from linkage with the Netherlands Cancer Registry and with hospital databases. Hazards ratios will be calculated for dietary and lifestyle factors at diagnosis in relation to recurrence and survival. Repeated measures analyses will be performed to assess changes over time in dietary and other factors in relation to recurrence and survival.
Fatty acid-inducible ANGPTL4 governs lipid metabolic response to exercise
Catoire, M. ; Alex, S. ; Paraskevopulos, N. ; Mattijssen, F.B.J. ; Mensink, M.R. ; Kersten, A.H. - \ 2014
Proceedings of the National Academy of Sciences of the United States of America 111 (2014)11. - ISSN 0027-8424 - p. E1043 - E1052.
human skeletal-muscle - angiopoietin-like protein-4 - lipoprotein-lipase - adipose-tissue - insulin-resistance - postprandial triacylglycerol - endurance exercise - in-vivo - expression - receptor
Physical activity increases energy metabolism in exercising muscle. Whether acute exercise elicits metabolic changes in nonexercising muscles remains unclear. We show that one of the few genes that is more highly induced in nonexercising muscle than in exercising human muscle during acute exercise encodes angiopoietin-like 4 (ANGPTL4), an inhibitor of lipoprotein lipase-mediated plasma triglyceride clearance. Using a combination of human, animal, and in vitro data, we show that induction of ANGPTL4 in nonexercising muscle is mediated by elevated plasma free fatty acids via peroxisome proliferator-activated receptor-d, presumably leading to reduced local uptake of plasma triglyceride-derived fatty acids and their sparing for use by exercising muscle. In contrast, the induction of ANGPTL4 in exercising muscle likely is counteracted via AMP-activated protein kinase (AMPK)-mediated down-regulation, promoting the use of plasma triglycerides as fuel for active muscles. Our data suggest that nonexercising muscle and the local regulation of ANGPTL4 via AMPK and free fatty acids have key roles in governing lipid homeostasis during exercise.
Increasing protein intake modulates lipid metabolism in healthy young men and women consuming a high-fat hypercaloric diet 1-3
Rietman, A. ; Schwarz, J. ; Blokker, B.A. ; Siebelink, E. ; Kok, F.J. ; Afman, L.A. ; Tome, D. ; Mensink, M.R. - \ 2014
The Journal of Nutrition 144 (2014)8. - ISSN 0022-3166 - p. 1174 - 1180.
energy-expenditure - hepatic steatosis - adipose-tissue - liver fat - quantification - disease - rats - homeostasis - accurate - insulin
The objective of this study was to evaluate the effect of increasing protein intake, at the expense of carbohydrates, on intrahepatic lipids (IHLs), circulating triglycerides (TGs), and body composition in healthy humans consuming a high-fat, hypercaloric diet. A crossover randomized trial with a parallel control group was performed. After a 2-wk run-in period, participants were assigned to either the control diet [n = 10; 27.8 energy percent (en%) fat, 16.9 en% protein, 55.3 en% carbohydrates] for 4 wk or a high-fat, hypercaloric diet (n = 17; >2 MJ/d) crossover trial with 2 periods of 2 wk, with either high-protein (HP) (37.7 en% fat, 25.7 en% protein, 36.6 en% carbohydrates) or normal-protein (NP) (39.4 en% fat, 15.4 en% protein, 45.2 en% carbohydrates) content. Measurements were performed after 2 wk of run-in (baseline), 2 wk of intervention (period 1), and 4 wk of intervention (period 2). A trend toward lower IHL and plasma TG concentrations during the HP condition compared with the NP condition was observed (IHL: 0.35 ± 0.04% vs. 0.51 ± 0.08%, P = 0.08; TG: 0.65 ± 0.03 vs. 0.77 ± 0.05 mmol/L, P = 0.07, for HP and NP, respectively). Fat mass was significantly lower (10.6 ± 1.72 vs. 10.9 ± 1.73 kg; P = 0.02) with the HP diet than with the NP diet, whereas fat-free mass was higher (55.7 ± 2.79 vs. 55.2 ± 2.80 kg; P = 0.003). This study indicated that an HP, high-fat, hypercaloric diet affects lipid metabolism. It tends to lower the IHL and circulating TG concentrations and significantly lowers fat mass and increases fat-free mass compared with an NP, high-fat, hypercaloric diet. This trail was registered at www.clinicaltrails.gov as NCT01354626.
Direct comparison of health effects by dietary polyphenols at equimolar doses in wildtype moderate high-fat fed C57BL/6JOlaHsd mice
Schothorst, E.M. van; Bunschoten, A. ; Hoevenaars, F.P.M. ; Stelt, I. van der; Janovska, P. ; Venema, D.P. ; Kopecky, J. ; Hollman, P.C.H. ; Keijer, J. - \ 2014
Food Research International 65 (2014)Part A. - ISSN 0963-9969 - p. 95 - 102.
adipose-tissue - body-weight - induced obesity - disease risk - quercetin - bioavailability - expression - flavonoids - leptin - (-)-epigallocatechin-3-gallate
Polyphenols generally show beneficial health effects upon supplementation in diet-induced obese rodent models, including reduced body weight gain and reduced levels of markers for cardiovascular diseases (CVD). However, there appear to be large differences between studies, which might be due to differences in models, strains, dietary background, or even concentration of polyphenol that is used. Therefore, we performed a systematic phenotypic evaluation of the effects of selected polyphenols in wildtype C57BL/6JOlaHsd mice. Epigallocatechin-gallate, quercetin, and resveratrol, representing three different phenolic classes, were each added in equimolar amounts (0.50% (w/w), 0.33%, and 0.25%, respectively) to a purified moderate high fat (30energy%) diet for 12 weeks. We studied the polyphenol-induced physiological and molecular effects between them and relative to the nonsupplemented control group during and at the end of the nutritional intervention. Results showed that these polyphenols were present in circulation, but did not induce beneficial health effects as analysed by oral glucose tolerance testing or serum adipokines and CVD-markers such as vascular adhesion molecules. Remarkably, transcriptomics of white adipose tissue showed overlapping sets of significantly differential transcript levels between these polyphenols; AMPK and Notch signalling were affected by these polyphenols. However, mitochondrial processes and mitochondrial density in this tissue did not differ between the polyphenols, which suggested that there was no direct effect on adipose tissue.
Thermoneutrality results in prominent diet-induced body weight differences in C57BL/6J mice, not paralleled by diet-induced metabolic differences
Hoevenaars, F.P.M. ; Bekkenkamp-Grovenstein, M. ; Janssen, R.J.R.J. ; Heil, S.G. ; Bunschoten, A. ; Hoek-van den Hil, E.F. ; Snaas-Alders, S.H. ; Teerds, K.J. ; Schothorst, E.M. van; Keijer, J. - \ 2014
Molecular Nutrition & Food Research 58 (2014)4. - ISSN 1613-4125 - p. 799 - 807.
adipose-tissue - mitochondrial-function - obesity - fat - thermogenesis - models - health - gene - induction - disease
Scope Mice are usually housed at 20–24°C. At thermoneutrality (28°C) larger diet-induced differences in obesity are seen. We tested whether this leads to large differences in metabolic health parameters. Methods and results We performed a 14-wk dietary intervention in C57BL/6J mice at 28°C and assessed adiposity and metabolic health parameters for a semipurified low fat (10 energy%) diet and a moderate high fat (30 energy%) diet. A large and significant diet-induced differential increase in body weight, adipose tissue mass, adipocyte size, serum leptin level, and, to some extent, cholesterol level was observed. No adipose tissue inflammation was seen. No differential effect of the diets on serum glucose, free fatty acids, triacylglycerides, insulin, adiponectin, resistin, PAI-1, MMP-9, sVCAM-1, sICAM-1, sE-selectin, IL-6, ApoE, fibrinogen levels, or HOMA index was observed. Also in muscle no differential effect on mitochondrial density, mitochondrial respiratory control ratio, or mRNA expression of metabolic genes was found. Finally, in liver no differential effect on weight, triacylglycerides level, aconitase/citrate synthase activity ratio was seen. Conclusion Low fat diet and moderate high fat diet induce prominent body weight differences at thermoneutrality, which is not paralleled by metabolic differences. Our data rather suggest that thermoneutrality alters metabolic homeostasis.
Omega-3 phospholipids from fish suppress hepatic steatosis by integrated inhibition of biosynthetic pathways in dietary obese mice
Rossmeisl, M. ; Medrikova, D. ; Schothorst, E.M. van; Pavlisova, J. ; Kuda, O. ; Hensler, M. ; Bardova, K. ; Flachs, P. ; Stankova, B. ; Vecka, M. ; Tvrizicka, E. ; Zak, A. ; Keijer, J. ; Kopecky, J. - \ 2014
Biochimica et Biophysica Acta. Molecular and Cell Biology of Lipids 1841 (2014)2. - ISSN 1388-1981 - p. 267 - 278.
polyunsaturated fatty-acids - long-chain omega-3-fatty-acids - krill oil supplementation - adipose-tissue - liver-disease - glucose-intolerance - insulin sensitivity - hepatobiliary axis - soybean lecithin - lipid-metabolism
Non-alcoholic fatty liver disease (NAFLD) accompanies obesity and insulin resistance. Recent meta-analysis suggested omega-3 polyunsaturated fatty acids DHA and EPA to decrease liver fat in NAFLD patients. Anti-inflammatory, hypolipidemic, and insulin-sensitizing effects of DHA/EPA depend on their lipid form, with marine phospholipids showing better efficacy than fish oils. We characterized the mechanisms underlying beneficial effects of DHA/EPA phospholipids, alone or combined with an antidiabetic drug, on hepatosteatosis. C57BL/6N mice were fed for 7 weeks an obesogenic high-fat diet (cHF) or cHF-based interventions: (i) cHF supplemented with phosphatidylcholine-rich concentrate from herring (replacing 10% of dietary lipids; PC), (ii) cHF containing rosiglitazone (10 mg/kg diet; R), or (iii) PC + R. Metabolic analyses, hepatic gene expression and lipidome profiling were performed. Results showed that PC and PC + R prevented cHF-induced weight gain and glucose intolerance, while all interventions reduced abdominal fat and plasma triacylglycerols. PC and PC + R also lowered hepatic and plasma cholesterol and reduced hepatosteatosis. Microarray analysis revealed integrated down-regulation of hepatic lipogenic and cholesterol biosynthesis pathways by PC, while R-induced lipogenesis was fully counteracted in PC + R. Gene expression changes in PC and PC + R were associated with preferential enrichment of hepatic phosphatidylcholine and phosphatidylethanolamine fractions by DHA/EPA. The complex down-regulation of hepatic lipogenic and cholesterol biosynthesis genes and the antisteatotic effects were unique to DHA/EPA-containing phospholipids, since they were absent in mice fed soy-derived phosphatidylcholine. Thus, inhibition of lipid and cholesterol biosynthesis associated with potent antisteatotic effects in the liver in response to DHA/EPA-containing phospholipids support their use in NAFLD prevention and treatment.
Tipping the inflammatory balance: inflammasome activation distinguishes metabolically unhealthy from healthy obesity
Stienstra, R. ; Stefan, N. - \ 2013
Diabetologia 56 (2013)11. - ISSN 0012-186X - p. 2343 - 2346.
induced insulin-resistance - adipose-tissue - fetuin-a - cells - accumulation - disease - humans - benign - tlr4
Expansion of adipose tissue mass, predominantly in the visceral depot, strongly associates with the development of metabolic complications that are often observed in obesity. In addition, in obesity, an increased prevalence of nonalcoholic fatty liver disease and reduced cardiorespiratory fitness are observed. However, not all obese individuals develop metabolic abnormalities. To better understand the molecular mechanisms that predispose obese humans to the development of metabolic diseases, comparing the metabolically healthy obese (MHO) vs an unhealthy obese phenotype (MUO) may be of great value. A new study by Esser et al (DOI:10.1007/s00125-013-3023-9) now provides important evidence that the MHO phenotype is associated with a lower activation of the NOD-like receptor family pyrin domain containing-3 (NLPR3) inflammasome in macrophages of visceral adipose tissue and a more favourable inflammatory profile as compared with the MUO phenotype. This finding could promote novel studies in humans to decipher stimuli and mechanisms leading to increased inflammasome activity, not only in adipose tissue, but also in other organs that are involved in the regulation of metabolism.
Caspase-1 deficiency in mice reduces intestinal triglyceride absorption and hepatic triglyceride secretion
Diepen, J.A. van; Stienstra, R. ; Vroegrijk, I.O.C.M. ; Berg, S.A.A. van den; Salvatori, D. ; Hooiveld, G.J.E.J. ; Kersten, A.H. ; Tack, C.J. ; Netea, M.G. ; Smit, J.W.A. ; Joosten, L.A.B. ; Havekes, L.M. ; Dijk, K.W. van; Rensen, P.C.N. - \ 2013
Journal of Lipid Research 54 (2013)2. - ISSN 0022-2275 - p. 448 - 456.
lipid-metabolism - adipose-tissue - fatty-acids - lipoprotein metabolism - insulin-resistance - immune-responses - inflammation - liver - inflammasomes - interleukin-1
Caspase-1 is known to activate the proinflammatory cytokines IL-1 beta and IL-18. Additionally, it can cleave other substrates, including proteins involved in metabolism. Recently, we showed that caspase-1 deficiency in mice strongly reduces high-fat diet-induced weight gain, at least partly caused by an increased energy production. Increased feces secretion by caspase-1-deficient mice suggests that lipid malabsorption possibly further reduces adipose tissue mass. In this study we investigated whether caspase-1 plays a role in triglyceride-(TG)-rich lipoprotein metabolism using caspase-1-deficient and wild-type mice. Caspase-1 deficiency reduced the postprandial TG response to an oral lipid load, whereas TG-derived fatty acid (FA) uptake by peripheral tissues was not affected, demonstrated by unaltered kinetics of [H-3] TG-labeled very low-density lipoprotein (VLDL)-like emulsion particles. An oral gavage of [H-3] TG-containing olive oil revealed that caspase-1 deficiency reduced TG absorption and subsequent uptake of TG-derived FA in liver, muscle, and adipose tissue. Similarly, despite an elevated hepatic TG content, caspase-1 deficiency reduced hepatic VLDL-TG production. Intestinal and hepatic gene expression analysis revealed that caspase-1 deficiency did not affect FA oxidation or FA uptake but rather reduced intracellular FA transport, thereby limiting lipid availability for the assembly and secretion of TG-rich lipoproteins.(jlr) The current study reveals a novel function for caspase-1, or caspase-1-cleaved substrates, in controlling intestinal TG absorption and hepatic TG secretion.-van Diepen, J. A., R. Stienstra, I. O. C. M. Vroegrijk, S. A. A. van den Berg, D. Salvatori, G. J. Hooiveld, S. Kersten, C. J. Tack, M. G. Netea, J. W. A. Smit, L. A. B. Joosten, L. M. Havekes, K. W. van Dijk, and P. C. N. Rensen. Caspase-1 deficiency in mice reduces intestinal triglyceride absorption and hepatic triglyceride secretion. J. Lipid Res. 2013. 54: 448-456.
Organ specificity of beta-carotene induced lung gene-expression changes in Bcmo 1-/- mice
Helden, Y.G.J. ; Godschalk, R.W.L. ; Schooten, F.J. van; Keijer, J. - \ 2013
Molecular Nutrition & Food Research 57 (2013)2. - ISSN 1613-4125 - p. 307 - 319.
nonsteroidal antiinflammatory drugs - triple-helix repeat - retinol efficacy trial - planar cell polarity - vitamin-a - cardiovascular-disease - adipose-tissue - epidemiologic evidence - cancer prevention - epithelial-cells
Scope - Whole genome transcriptome analysis of male and female beta-carotene 15,15'-monooxygenase knockout (Bcmo1-/-) and Bcmo1+/+ (wild-type) mice with or without 14 wk of BC supplementation was done. We previously showed that only 1.8% of the genes regulated by BC in lung were also regulated in liver and inguinal white adipose tissue (iWAT), suggesting lung specific responses. Here, we explicitly questioned the lung specificity. Methods and results - We show that BC supplementation resulted in an opposite direction of gene-regulation in male compared to female Bcmo1-/- mice in lung, liver, and iWAT. This supports a systemic effect of BC on steroid hormone metabolism mediated responses. Lung, liver, and iWAT of female Bcmo1-/- mice showed an increased inflammatory response, which was counteracted by supplementation of BC. This supports a genotype dependent increased sensitivity of female mice for vitamin A deficiency. Finally, the effect of BC on Wnt signaling in male Bcmo1-/- mice was examined. Frizzled homolog 6 (Fzd6) downregulation was seen in all three tissues. Collagen triple helix containing 1 (Cthrc1) downregulation was seen in lung tissue only, suggesting specificity. Upregulation of genes involved in oxygen sensing was seen in lung and iWAT, while protocadherin upregulation was only seen in lung. Conclusion - Our results demonstrate that effects of BC are strongly sex dependent. While effects of BC on hormone metabolism mediated responses and inflammation are systemic, effects on Wnt signaling may be lung specific.
Correlation between activation of PPAR¿ and resistin downregulation in a mouse adipocyte cell line by a series of thiazolidinediones.
Sotiriou, A. ; Blaauw, R.H. ; Meijer, C. ; Gijsbers, L.H. ; Burg, B. van der; Vervoort, J. ; Rietjens, I.M.C.M. - \ 2013
Toxicology in Vitro 27 (2013)5. - ISSN 0887-2333 - p. 1425 - 1432.
insulin-resistance - adipose-tissue - antihyperglycemic agents - expression - receptor - glucose - metabolism - mechanisms - biology - obesity
The present study shows significant correlations between the EC50 for PPAR¿ activation in a reporter gene cell line and resistin downregulation in mouse adipocytes, and between the IC50 for resistin downregulation and the already published minimum effective dose for antihyperglycemic activity in a mouse model. These correlations indicate that PPAR¿ mediated downregulation of resistin might promote insulin sensitivity and that downregulation of resistin in mouse adipocytes provides an adequate and possibly more direct bioassay for screening of newly developed antihyperglycemic compounds. Because of the higher throughput of the PPAR¿ the resistin downregulation assays seems most suitable to be used as a second tier in a tiered screening strategy.
Activated complement factor 3 is associated with liver fat and liver enzymes: the CODAM study
Wlazlo, N. ; Greevenbroek, M.M.J. van; Ferreira, I. ; Jansen, E.H.J.M. ; Feskens, E.J.M. ; Kallen, C.J.H. van der; Schalkwijk, C.G. ; Bravenboer, B. ; Stehouwer, C.D.A. - \ 2013
European Journal of Clinical Investigation 43 (2013)7. - ISSN 0014-2972 - p. 679 - 688.
insulin-resistance - nonalcoholic steatohepatitis - alanine aminotransferase - medical progress - adipose-tissue - disease - c3 - mice - cytokines - protein
Background The complement system may be involved in the pathogenesis of alcoholic and nonalcoholic liver disease, although studies in humans are scarce. For this reason, we investigated whether circulating levels of activated complement factor 3 (C3a) were associated with hepatic steatosis and hepatocellular damage. Materials and methods Plasma C3a, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT) were determined in 523 individuals (61% men, age 59 ± 7 years). Liver enzymes (LEs) were standardized and compiled into a LE score. Liver fat content was estimated using a predictive equation that has recently been validated with magnetic resonance spectrometry. Cross-sectional associations between C3a and liver fat or LE s were investigated with multiple linear regression analyses, stratified in no-to-moderate vs. heavy alcohol consumers (men: > 30 g/day; women: > 20 g/day). Results C3a was associated with liver fat percentage both in the no-to-moderate (ß = 0·223; 95%CI 0·036; 0·409) and in the heavy alcohol consumers (ß = 0·632; 95%CI 0·259–1·004; P-interaction = 0·047). C3a was also associated with the LE score in heavy alcohol consumers (ß = 0·917; 95%CI 0·443–1·392), but not in no-to-moderate alcohol consumers (ß = 0·042; 95%CI -0·198 to 0·281; P-interaction = 0·001). Conclusions C3a levels, as a marker of complement activation, were associated with liver fat content and hepatocellular injury, at least in subjects who consume considerable amounts of alcohol daily.