Jonatan Miranda

Salt-inducible kinase 2 links transcriptional coactivator p300 phosphorylation to the prevention of ChREBP-dependent hepatic steatosis in mice

Obesity and type 2 diabetes are associated with increased lipogenesis in the liver. This results in fat accumulation in hepatocytes, a condition known as hepatic steatosis, which is a form of nonalcoholic fatty liver disease (NAFLD), the most common cause of liver dysfunction in the United States. Carbohydrate-responsive element-binding protein (ChREBP), a transcriptional activator of glycolytic and lipogenic genes, has emerged as a major player in the development of hepatic steatosis in mice. However, the molecular mechanisms enhancing its transcriptional activity remain largely unknown. In this study, we have identified the histone acetyltransferase (HAT) coactivator p300 and serine/threonine kinase salt-inducible kinase 2 (SIK2) as key upstream regulators of ChREBP activity. In cultured mouse hepatocytes, we showed that glucose-activated p300 acetylated ChREBP on Lys672 and increased its transcriptional activity by enhancing its recruitment to its target gene promoters. SIK2 inhibited p300 HAT activity by direct phosphorylation on Ser89, which in turn decreased ChREBP-mediated lipogenesis in hepatocytes and mice overexpressing SIK2. Moreover, both liver-specific SIK2 knockdown and p300 overexpression resulted in hepatic steatosis, insulin resistance, and inflammation, phenotypes reversed by SIK2/p300 co-overexpression. Finally, in mouse models of type 2 diabetes and obesity, low SIK2 activity was associated with increased p300 HAT activity, ChREBP hyperacetylation, and hepatic steatosis. Our findings suggest that inhibition of hepatic p300 activity may be beneficial for treating hepatic steatosis in obesity and type 2 diabetes and identify SIK2 activators and specific p300 inhibitors as potential targets for pharmaceutical intervention.

Nutritional Differences Between a Gluten-free Diet and a Diet Containing Equivalent Products with Gluten

The gluten-free (GF) products market represents one of the most prosperous markets in the field of food and beverages in the immediate future. Historically, counselling for celiac disease has focused on the absence of gluten in foods, however the nutritional quality of GF foodstuffs is an important aspect to consider. The aim of the present work was to compare the nutritional composition of the 206 GF rendered products most consumed in Spain, against the composition of 289 equivalent foods with gluten, and to make a comparison between the diet including GF products and the same diet with equivalent products with gluten in a 58 adult celiac population. The results of the present collaborative study pointed out differences in calorie, macronutrient, fiber, sodium, salt and cholesterol content between GF rendered and gluten-containing foodstuffs. Thus, calorie and nutrient intake in a GF diet is different when compared to its equivalent diet with gluten. Following a diet based on GF products could suppose a nutritional imbalance for celiac patients as well as for non-celiacs who follow a diet that includes many GF rendered foodstuffs.

Hepatic lipid metabolic pathways modified by resveratrol in rats fed an obesogenic diet.

OBJECTIVE The scientific community is on the look-out for safe biomolecules useful in the prevention of obesity and related aberrations such as fatty liver. This study analyzed the influence of resveratrol on hepatic triacylglycerol metabolism. METHODS Male Sprague-Dawley rats were divided into control and resveratrol-treated groups (30 mg/kg of body weight per day) and fed a commercial obesogenic diet for 6 wk. Liver triacylglycerol content and the activity of carnitine palmitoyl transferase-Ia (CPT-Ia), acyl-coenzyme A oxydase (ACO), fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme (ME), acetyl-coenzyme A carboxylase (ACC), adenosine monophosphate-activated protein kinase (AMPK), and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) activation were measured. Mitochondrial protein cytochrome C oxidase subunit 2 (COXII), mitochondrial transcription factor A (TFAM), sterol regulatory element-binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-α (PPAR-α), sirtuin-1 (SIRT1), hepatocyte nuclear factor receptor-4α (HNF-4α), and PGC-1α mRNA levels were also analyzed. Serum insulin was quantified. RESULTS Resveratrol decreased liver fat accumulation, increased CPT-Ia and ACO, and decreased ACC activities. Other lipogenic enzymes, FAS, ME, and G6PDH were not modified. The polyphenol activated AMPK and PGC-1α. The expression of SRBP-1c, PPAR-α, SIRT1, PGC-1α, HNF-4α, TFAM, and COXII was not modified. No changes in serum insulin levels were observed. CONCLUSION Resveratrol partly prevents the increase in liver fat accumulation induced by high-fat high-sucrose feeding by increasing fatty acid oxidation and decreasing lipogenesis. These effects are mediated by the activation of the AMPK/SIRT1 axis.

Thermogenesis is involved in the body-fat lowering effects of resveratrol in rats.

The effect of resveratrol on thermogenesis in skeletal muscle and interscapular brown adipose tissue (IBAT) was investigated. Rats were fed an obesogenic diet supplemented with resveratrol (30mg/kg/day) or not supplemented for 6weeks. Resveratrol intake led to increased gene expression of mitochondrial-transcription-factor-A (TFAM), mitochondrial-protein-cytochrome-C-oxidase subunit-2 (COX2), sirtuin-1 (SIRT1), peroxisome-proliferator-activated-receptor-β/δ (PPARβ/δ) and proliferator-activated-receptor-gamma-coactivator1-α (PGC-1α) in IBAT and increased UCP1protein expression; however, peroxisome-proliferator-activated-receptor-α (PPARα) expression remained unchanged. In gastrocnemius muscle, resveratrol increased the gene expression of TFAM and COX2; however, no changes were observed in levels of SIRT1, PGC-1α and PPARβ/δ. Acetylated-PGC-1α was decreased in the resveratrol-treated group, indicating a higher level of activation, and a significant increase of UCP3 protein expression was observed in this group. The increases in UCP protein expression in two important thermogenic tissues after resveratrol treatment may contribute to increased whole-body energy dissipation, which may help to better understand the body-fat lowering effect of this polyphenol.

High-throughput sequencing of microRNAs in peripheral blood mononuclear cells: identification of potential weight loss biomarkers.

Introduction MicroRNAs (miRNAs) are being increasingly studied in relation to energy metabolism and body composition homeostasis. Indeed, the quantitative analysis of miRNAs expression in different adiposity conditions may contribute to understand the intimate mechanisms participating in body weight control and to find new biomarkers with diagnostic or prognostic value in obesity management. Objective The aim of this study was the search for miRNAs in blood cells whose expression could be used as prognostic biomarkers of weight loss. Methods Ten Caucasian obese women were selected among the participants in a weight-loss trial that consisted in following an energy-restricted treatment. Weight loss was considered unsuccessful when <5% of initial body weight (non-responders) and successful when >5% (responders). At baseline, total miRNA isolated from peripheral blood mononuclear cells (PBMC) was sequenced with SOLiD v4. The miRNA sequencing data were validated by RT-PCR. Results Differential baseline expression of several miRNAs was found between responders and non-responders. Two miRNAs were up-regulated in the non-responder group (mir-935 and mir-4772) and three others were down-regulated (mir-223, mir-224 and mir-376b). Both mir-935 and mir-4772 showed relevant associations with the magnitude of weight loss, although the expression of other transcripts (mir-874, mir-199b, mir-766, mir-589 and mir-148b) also correlated with weight loss. Conclusions This research addresses the use of high-throughput sequencing technologies in the search for miRNA expression biomarkers in obesity, by determining the miRNA transcriptome of PBMC. Basal expression of different miRNAs, particularly mir-935 and mir-4772, could be prognostic biomarkers and may forecast the response to a hypocaloric diet.

Association between dietary phylloquinone intake and peripheral metabolic risk markers related to insulin resistance and diabetes in elderly subjects at high cardiovascular risk

BackgroundVitamin K has been related to glucose metabolism, insulin sensitivity and diabetes. Because inflammation underlies all these metabolic conditions, it is plausible that the potential role of vitamin K in glucose metabolism occurs through the modulation of cytokines and related molecules. The purpose of the study was to assess the associations between dietary intake of vitamin K and peripheral adipokines and other metabolic risk markers related to insulin resistance and type 2 diabetes mellitus.MethodsCross-sectional and longitudinal assessments of these associations in 510 elderly participants recruited in the PREDIMED centers of Reus and Barcelona (Spain). We determined 1-year changes in dietary phylloquinone intake estimated by food frequency questionnaires, serum inflammatory cytokines and other metabolic risk markers.ResultsIn the cross-sectional analysis at baseline no significant associations were found between dietary phylloquinone intake and the rest of metabolic risk markers evaluated, with exception of a negative association with plasminogen activator inhibitor-1. After 1-year of follow-up, subjects in the upper tertile of changes in dietary phylloquinone intake showed a greater reduction in ghrelin (−15.0%), glucose-dependent insulinotropic peptide (−12.9%), glucagon-like peptide-1 (−17.6%), IL-6 (−27.9%), leptin (−10.3%), TNF (−26.9%) and visfatin (−24.9%) plasma concentrations than those in the lowest tertile (all p<0.05).ConclusionThese results show that dietary phylloquinone intake is associated with an improvement of cytokines and other markers related to insulin resistance and diabetes, thus extending the potential protection by dietary phylloquinone on chronic inflammatory diseases.Trial registrationhttp://www.controlled-trials.com as ISRCTN35739639

A comparison between CLNA and CLA effects on body fat, serum parameters and liver composition

The potential of conjugated linoleic acid (CLA) as an anti-obesity molecule for humans is still a matter for debate. Thus, a great deal of scientific work is focussed on the research of new effective molecules without deleterious effects on health. The aim of the present work was to analyse the effects of jacaranda seed oil, rich in a conjugated linolenic acid (CLNA), jacaric acid (cis-8,trans-10,cis-12), on body fat, serum parameters and liver composition in rats, and to compare these effects with those oftrans-10,cis-12 CLA. Twenty-six male Wistar rats were divided into three groups fed with high-fat diets, supplemented or not (control group) with 0.5%trans-10,cis-12 CLA (CLA group) or 0.5% jacaric acid (CLNA group) for 7 weeks. No statistical differences in food intake or in final body weight were found. Whereas CLA reduced adipose tissue size, CLNA did not. Both CLA and CLNA significantly reduced non-HDL-cholesterol. In spite of a lack of significant changes in glucose and insulin levels, HOMA-IR index was significantly increased, as well as did non-esterified fatty acid levels in CLNA-fed rats. No changes in liver composition were observed. In conclusion, under our experimental conditions, jacaric acid, unlike CLA, does not show a body-fat lowering effect. Even though it leads to a healthy lipoprotein profile, it impairs insulin function. Consequently, it cannot be proposed as an anti-obesity molecule.ResumenEl potencial del ácido linoleico conjugado (CLA) como molécula anti-obesidad para seres humanos sigue siendo una cuestión en debate. Por ello, gran cantidad de trabajos científicos se centra en la investigación de nuevas moléculas eficaces y sin efectos nocivos sobre la salud. El objectivo del presente trabajo fue estudiar, en rata, los efectors del aceite de semillas de jacaranda, rico en un ácido linolénico conjugado (CLNA), el ácido jacárico (cis-8,trans-10,cis-12), sobre la grasa corporal, parámetros séricos y la composición del hígado, y comparar estos efectos con los deltrans-10,cis-12, CLA. Se utilizaron 26 ratas Wistar macho divididas en tres grupos que fueron alimentados durante 7 semanas con dietas hipergrasas, suplementadas o no (grupo control) al 0,5% con eltrans-10,cis-12 CLA (grupo CLA) o al 0,5% con el ácido jacárico (grupo CLNA). No se encontraron diferencias significativas en la ingesta de dieta, ni en el peso corporal final, ni en la composición del hígado. El CLA redujo la masa adiposa, pero no lo hizo el CLNA. Ambos disminuyeron significativamente el colesterol no-HDL. A pesar de la ausencia de cambios significativos en la glucemia e insulinemia, el índice HOMA-IR y los niveles séricos de AGL aumentaron significativamente en las ratas alimentadas con CLNA. En conclusión, en nuestras condiciones experimentales, el ácido jacárico, a diferencia del CLA, no muestra un efecto reductor de la grasa corporal. A pesar de que mejora el perfil de lipoproteínas, altera la función insulínica. Por lo tanto, este CLNA no puede ser propuesto como una molécula antiobesidad.

Doses of Quercetin in the Range of Serum Concentrations Exert Delipidating Effects in 3T3-L1 Preadipocytes by Acting on Different Stages of Adipogenesis, but Not in Mature Adipocytes

Scope. To determine whether doses of quercetin in the range of serum concentrations exert any effect on triacylglycerol accumulation in maturing preadipocytes and mature adipocytes. The influence on the expression of adipogenic markers as well as on gene expression and activity of enzymes involved in triacylglycerol metabolism were assessed. Methods and Results. 3T3-L1 preadipocytes were treated during differentiation and mature adipocytes for 24 hours with low doses (0.1–10 µM) of quercetin. Triacylglycerol content in both cell types and free fatty acid and glycerol in the incubation medium of mature adipocytes were measured spectrophotometrically. Gene and protein expression were assessed by RT-PCR and Western blot. LPL and FAS activities were quantified. During differentiation quercetin reduced triacylglycerol content at doses from 0.5 to 10 µM. 1 µM of quercetin reduced C/EBPβ gene expression, SREBP1 mature protein levels, and PPARγ gene expression. 10 µM of quercetin reduced LPL gene expression and PPARγ and SREBP1c expression. In mature adipocytes, only 10 µM of quercetin reduced triacylglycerol content. Lipogenic FAS expression and activity were reduced at this dose. Conclusion. Quercetin, in the range of serum concentrations, is able to inhibit adipogenesis, but higher doses, at least 10 µM, are needed to reduce fat accumulation in mature adipocytes.

Effects of trans-10,cis-12 conjugated linoleic acid on body fat and serum lipids in young and adult hamsters.

The aim of the present work was to determine whether t-10, c-12 conjugated linoleic acid (CLA) feeding was able to reduce body fat accumulation and improve the serum lipid profile in adult hamsters fed an atherogenic diet, in order to compare these effects with those observed in young growing hamsters. Young and adult hamsters were fed semi-purified atherogenic diets supplemented with 0.5% linoleic acid or 0.5% t-10, c-12 CLA for 6 weeks. Body weight and food intake were measured every two days. Adipose tissue from different anatomical locations, liver and gastrocnemious muscle were dissected and weighed. Cholesterol, triacylglycerols, non-esterified fatty acids and proteins were determined spectrophotometrically and water content by gravimetry. In young hamsters, no significant differences were found in food intake, final body weight and gastrocnemious muscle weight. White adipose tissue weights were reduced, liver weight was increased and cholesterol and triacylglycerols in both serum and liver were reduced. In adult hamsters, CLA feeding decreased food intake and adipose tissue weights. No changes were observed in other parameters. The present study demonstrates that age has an influence in hamster responsiveness to t-10, c-12 CLA because, although when this isomer is added to an atherogenic diet it reduces body fat accumulation in both young and adults hamsters, the lessening of the effects on serum lipids brought about by atherogenic feeding is only observed in young animals. Moreover, it is clear that liver is a target for CLA in young but not in adult hamsters.ResumenEl objetivo del presente estudio fue determinar si el isómero t-10, c-12 del ácido linoleico conjugado (ALC) era capaz de reducir la acumulación de grasa corporal y de mejorar el perfil lipídico en hámsteres adultos alimentados con una dieta aterogénica, con el fin de compararlos con los observados en hámsteres jóvenes en crecimiento. Los animales se alimentaron con dietas aterogénicas suplementadas con 0,5% de ácido linoleico ó 0,5% de ALC t-10, c-12 durante 6 semanas. Se midió cada dos días la ingesta de alimento y el peso corporal. Se diseccionaron y pesaron tejidos adiposos de diferentes localizaciones anatómicas, el hígado y los dos músculos gastrocnemios. El colesterol, los triglicéridos, los ácidos grasos libres y las proteínas se valoraron espectrofotométricamente ricamente y el agua por gravimetría. En los animales jóvenes no se observaron diferencias significativas en la ingesta, el peso corporal final y el peso de los músculos gastrocnemios. Los pesos de los tejidos adiposos blancos se redujeron, el peso de hígado aumentó y el colesterol y los triglicéridos disminuyeron, tanto en suero como en higado. En hámsteres adultos, el ALC disminuyó la ingesta y los pesos de los tejidos adiposos, pero no se observaron cambios en los demás parámetros. El presente estudio demuestra que la edad influye en la respuesta del hámster al ALC t-10, c-12 porque, aunque al ser anadido a una dieta aterogénica reduce la grasa corporal tanto en animales jóvenes como adultos, la atenuación de los efectos de esta dieta sobre los lípidos séricos sólo se pone de manifiesto en los jóvenes. Además, sólo en estos últimos, el hígado es claramente una diana para el ALC.

A combination of resveratrol and quercetin induces browning in white adipose tissue of rats fed an obesogenic diet

To analyze whether a combination of quercetin (Q) and resveratrol (RSV) would induce a white adipose tissue (WAT) browning effect.