Diet-induced maternal obesity alters insulin signalling in male mice offspring rechallenged with a high-fat diet in adulthood

Modern lifestyle has resulted in an increase in the prevalence of obesity and its comorbidities in pregnant women and the young population. It has been well established that the consumption of a high-fat diet (HFD) has many direct effects on glucose metab118122FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2011/22156-7; 2013/12003-

Prolonged fasting elicits increased hepatic triglyceride accumulation in rats born to dexamethasone-treated mothers

We investigated the effect of dexamethasone during the last week of pregnancy on glucose and lipid metabolism in male offspring. Twelve-week old offspring were evaluated after fasting for 12-hours (physiological) and 60-hours (prolonged). Physiological fasting resulted in glucose intolerance, decreased glucose clearance after pyruvate load and increased PEPCK expression in rats born to dexamethasone-treated mothers (DEX). Prolonged fasting resulted in increased glucose tolerance and increased glucose clearance after pyruvate load in DEX. These modulations were accompanied by accumulation of hepatic triglycerides (TG). Sixty-hour fasted DEX also showed increased citrate synthase (CS) activity, ATP citrate lyase (ACLY) content, and pyruvate kinase 2 (pkm2), glucose transporter 1 (slc2a1) and lactate dehydrogenase-a (ldha) expressions. Hepatic AKT2 was increased in 60-hour fasted DEX, in parallel with reduced miRNAs targeting the AKT2 gene. Altogether, we show that metabolic programming by prenatal dexamethasone is characterized by an unexpected hepatic TG accumulation during prolonged fasting. The underlying mechanism may depend on increased hepatic glycolytic flux due to increased pkm2 expression and consequent conversion of pyruvate to non-esterified fatty acid synthesis due to increased CS activity and ACLY levels. Upregulation of AKT2 due to reduced miRNAs may serve as a permanent mechanism leading to increased pkm2 expression.Sao Paulo Research Foundation (FAPESP)National Counsel of Technological and Scientific Development (CNPq)Coordination for the Improvement of Higher Level or Education Personnel (CAPES)Univ Sao Paulo, Inst Biomed Sci, Dept Physiol & Biophys, Sao Paulo, BrazilUniv Estadual Campinas, Fac Med Sci, Dept Pharmacol, Campinas, SP, BrazilUniv Fed Sao Paulo, Inst Environm Chem & Pharmaceut Sci, Diadema, BrazilUniv Fed Sao Paulo, Inst Environm Chem & Pharmaceut Sci, Diadema, BrazilWeb of Scienc

Changes in PGC-1α-Dependent Mitochondrial Biogenesis Are Associated with Inflexible Hepatic Energy Metabolism in the Offspring Born to Dexamethasone-Treated Mothers

In the present study we investigated the participation of hepatic peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α) in the metabolic programming of newborn rats exposed in utero to dexamethasone (DEX). On the 21st day of life, fasted offspring born to DEX-treated mothers displayed increased conversion of pyruvate into glucose with simultaneous upregulation of PEPCK (phosphoenolpyruvate carboxykinase) and G6Pase (glucose-6-phosphatase). Increased oxidative phosphorylation, higher ATP/ADP ratio and mitochondrial biogenesis and lower pyruvate levels were also found in the progeny of DEX-treated mothers. On the other hand, the 21-day-old progeny of DEX-treated mothers had increased hepatic triglycerides (TAG) and lower CPT-1 activity when subjected to short-term fasting. At the mechanistic level, rats exposed in utero to DEX exhibited increased hepatic PGC-1α protein content with lower miR-29a-c expression. Increased PGC-1α content was concurrent with increased association to HNF-4α and NRF1 and reduced PPARα expression. The data presented herein reveal that changes in the transcription machinery in neonatal liver of rats born to DEX-treated mothers leads to an inflexible metabolic response to fasting. Such programming is hallmarked by increased oxidative phosphorylation of pyruvate with impaired FFA oxidation and hepatic TAG accumulation.</jats:p

Prenatal exposure to dexamethasone exacerbates the effects of fructose on hepatic lipid metabolism.

A reprogramação fenotípica causada por alterações ambientais distintas associada ao alto consumo de frutose na idade adulta poderia contribuir para o agravamento de distúrbios metabólicos. Para avaliar se a exposição pré-natal à dexametasona (DEX) modula os efeitos do consumo de frutose sobre o metabolismo hepático, investigamos a prole masculina nascida de ratas Wistar tratadas ou não com DEX (0,2mg/kg massa corpórea/dia) no 3°período gestacional. As proles de mãe controle (sem tratamento) e mães tratadas com DEX foram mantidas sem frutose (CTL e DEX) ou foram suplementadas com solução de frutose a 10% durante 8 semanas (frutose e DEX+frutose). Tanto os grupos frutose quanto DEX+frutose apresentaram consumo de frutose semelhante, intolerância à glicose e aumento da atividade máxima da enzima glicolítica PFK. Apenas DEX+frutose apresentou aumento dos triglicérides hepáticos e acúmulo de lipídeos intra-hepáticos. Além disso, o grupo DEX+frutose não mostrou expressão aumentada dos genes sec22, mttp e apoB, envolvidos na síntese, montagem e secreção de VLDL; paralelamente, observamos diminuição na concentração de triglicérides em 6 horas após injeção de Tyloxapol comparado ao DEX e frutose. Ainda, apresentou diminuição da expressão de BECLIN1, da expressão gênica de HSP90, e alteração de marcadores moleculares relacionados ao HCC, como diminuição de TP53, P21 e TIGAR, e aumento da expressão de mRNA afp e hsp70. Em conjunto, nossos dados indicam que a exposição à DEX in útero leva a uma resposta metabólica única frente a alta ingestão de frutose na vida adulta. O aumento da gliconeogênese e intolerância à glicose, a redução na produção e secreção de VLDL e o prejuízo no fluxo autofágico estimulam o acúmulo de gotículas lipídicas no fígado que pode favorecer o desenvolvimento de HCC em resposta à alta ingestão de frutose.Phenotypic programming caused by distinct environmental changes and associated with high fructose consumption in adulthood could contribute to the worsening of metabolic disorders. To evaluate whether prenatal exposure to Dexamethasone (DEX) modulates the effects of fructose consumption on hepatic metabolism, we investigated male offspring born to Wistar rats treated with or without DEX (0.2 mg / kg body weight / day) at 3rd gestational period. The offspring of control (untreated) and DEX treated mothers were maintained without fructose (CTL and DEX) or supplemented with 10% fructose solution for 8 weeks (fructose and DEX+fructose). Both the fructose and DEX+fructose groups showed similar fructose consumption, glucose intolerance and increased maximal activity of the glycolytic enzyme PFK. Only DEX+fructose presented increase in hepatic triglycerides and accumulation of intrahepatic lipids. In addition, the DEX + fructose group did not show increased expression of sec22, mttp and apoB genes involved in the synthesis, assembly and secretion of VLDL; in parallel, we observed a decrease in TG concentration in 6 hours after injection of Tyloxapol, compared to DEX and fructose. In addition, there was a decrease in BECLIN1 expression, HSP90 gene expression, and alteration of HCC-related molecular markers such as TP53, P21 and TIGAR, and increased expression of afp and hsp70 mRNA. Taken together, our data indicate that exposure to DEX in utero leads to a unique metabolic response to high fructose intake in adult life. Increased gluconeogenesis and glucose intolerance, reduction in VLDL production and secretion, and loss of autophagic flow stimulate the accumulation of lipid droplets in the liver, that may favor the development of HCC in response to high fructose intake

Modulation of cholinergic anti-inflammatory pathway in the liver and white adipose tissue of the offspring by maternal high fat diet consumption

Orientadores: Márcio Alberto Torsoni, Marciane MilanskiDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências AplicadasResumo: A obesidade materna está relacionada ao desenvolvimento de distúrbios metabólicos na prole. O processo inflamatório desencadeado pelo consumo materno de dieta rica em gordura parece envolver a ativação da via TLR4/NFkB. O reflexo anti-inflamatório colinérgico é ativado por meio do nervo vago e através da subunidade ?7 dos receptores nicotínicos de acetilcolina (?7nAChR) que reduz a ativação do NF-kB e diminui a produção de citocinas inflamórias e a intensidade da resposta imune. O bloqueio destes receptores ou a denervação da via eferente provoca intensa produção de mediadores inflamatórios em resposta ao lipopolissacarídeo (LPS). Neste sentido, estudos com modelos experimentais têm mostrado que a prole de mães obesas apresenta a resposta inflamatória exacerbada após um desafio com (LPS) ou dieta hiperlipídica (HFD). Assim, é possível que estes animais apresentem o controle do ajuste fino da resposta inflamatória afetado pelo consumo materno de HFD. O objetivo central foi avaliar a funcionalidade da via anti-inflamatória colinérgica (AIC) na prole de mães obesas. Para isso prole de camundongos swiss machos com 28 dias de vida (d28) obtidos de mães alimentadas com dieta HFD (HFD-O) ou dieta controle (SC-O) durante a gestação e lactação foram empregados no estudo. O LPS e a nicotina foram administrado via intraperitoneal (IP) e intracerebroventricular (i.c.v), respectivamente, e a atividade da via foi avaliada. Os resultados mostram repressão nas proteínas chaves envolvidas na via AIC no fígado e tecido adiposo branco (WAT), elevado nível sérico de TNF-? e redução dos marcadores para macrófago do tipo M2 (Chil 3, Arg1 e IL10) no grupo HFD-O. O ?7nAChR foi detectado nas células do sistema imunitário (colocalizado com F4/80) e adipócitos de camundongos SC-O, não sendo obeservada a expressão do receptor nos adipócitos do grupo HFD-O. Além disso, foi obeservado reduzida associação entre STAT3 e NF-kB no grupo HFD-O em ambos os tecidos após estímulo de nicotina, maior atividade da acetilcolinesterase basal no grupo HFD-O e aumento da expressão de TNF-? e IL-1? no fígado e WAT de camundongos após estímulo com LPS, sendo este efeito ainda maior no grupo HFD-O. Assim podemos concluir que a obesidade materna e o consumo de HFD durante a gestação e lactação promove adaptações moleculares na via AIC e estas podem levar aos distúrbios metabólicos associados a supernutrição no período perinatalAbstract: Maternal obesity is related to the development of metabolic disorders in offspring. The inflammatory process triggered by maternal consumption of high-fat diet appears to be triggered by activation of the TLR4/NFkB. The cholinergic anti-inflammatory reflex is activated by the vagus nerve and by the ?7 subunit of the nicotinic acetylcholine receptors (?7nAChR) reduces the activation of NF-kB and consequently decreases the production of TNF? and intensity of the immune response. Blockade of these receptors or efferent causes denervation of intense production of cytokines in response to lipopolysaccharide (LPS). In this regard, studies on experimental models have shown that obese offspring of mothers shows the increased inflammatory response following a challenge with LPS or (HFD). Thus, it is possible that these animals have control of fine-tuning the inflammatory response affected by HFD maternal consumption. The main objective was to evaluate the cholinergic anti-inflammatory pathway functionality (AIC) in the offspring of obese mothers. For this offspring of swiss males with 28 days old mice (d28) obtained from mothers fed with HFD diet (HFD-O) or control diet (SC-O) during pregnancy and lactation were used in the study. The LPS and nicotine were administered intraperitoneally (IP) and intracerebroventricular (i.c.v), respectively, and the track activity was evaluated. The results show repression in key protein involved in the cholinergic anti-inflammatory pathway in the liver and white adipose tissue (WAT), high serum levels of TNF-? and reducing markers M2 type macrophage (Chil 3, Arg1 e IL10) in the group HFD-O. The ?7nAChR was detected in immune system cells (co-located with F4/80) and adipocytes of group SC-O, not being obeserved receptor expression in adipocytes of HFD-O group. Furthermore, it was observed reduced association between STAT3 and NFkB in group HFD-O in both tissues after nicotine stimulation, basal activity greater acetylcholinesterase in the HFD-O and increased expression of TNF-? and IL-1? in liver and WAT of mice after stimulation with LPS, being this effect even greater in HFD-O. So we can conclude that maternal obesity and the consumption of HFD during pregnancy and lactation promotes molecular adaptations towards cholinergic pathway and these can lead to metabolic disorders associated with overnutrition in the perinatal periodMestradoMetabolismo e Biologia MolecularMestra em Ciências da Nutrição e do Esporte e Metabolism

High-fat Diet During Pregnancy And Lactation Impairs The Cholinergic Anti-inflammatory Pathway In The Liver And White Adipose Tissue Of Mouse Offspring

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Cholinergic anti-inflammatory pathway (CAP) prevents inflammatory cytokines production. The main was to evaluate the effect of maternal obesity on cholinergic pathway in the offspring. Female mice were subjected to either standard chow (SC) or high-fat diet (HFD) during pregnancy and the lactation period. After weaning, only male offspring from HFD dams (HFD-O) and from SC dams (SC-O) were fed the SC diet. Key proteins of the CAP were downregulated and serum TNF-alpha was elevated in the HFD-O mice. STAT3 and NF-kappa B activation in HFD-O mice ICV injected with nicotine (agonist) were lower than SC-O mice. Basal cholinesterase activity was upregulated in HFD-O mice in both investigated tissues. Lipopolysaccharide increased TNF-alpha and IL-1 beta expression in the liver and WAT of SC-O mice, but this effect was greater in HFD-O mice. In conclusion these changes exacerbated cytokine production in response to LPS and contributed to the reduced sensitivity of the CAP. (C) 2015 Published by Elsevier Ireland Ltd.422C192202Sao Paulo Research Foundation (FAPESP) [14/18165-9, 13/10706-8]CNPq [476643/2012-0]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

High-fat diet during pregnancy and lactation impairs the cholinergic anti-inflammatory pathway in the liver and white adipose tissue of mouse offspring

14/18165-9; 13/10706-8476643/2012-0Cholinergic anti-inflammatory pathway (CAP) prevents inflammatory cytokines production. The main was to evaluate the effect of maternal obesity on cholinergic pathway in the offspring. Female mice were subjected to either standard chow (SC) or high-fat diet (HFD) during pregnancy and the lactation period. After weaning, only male offspring from HFD dams (HFD-O) and from SC dams (SC-O) were fed the SC diet. Key proteins of the CAP were downregulated and serum TNF-α was elevated in the HFD-O mice. STAT3 and NF-κB activation in HFD-O mice ICV injected with nicotine (agonist) were lower than SC-O mice. Basal cholinesterase activity was upregulated in HFD-O mice in both investigated tissues. Lipopolysaccharide increased TNF-α and IL-1β expression in the liver and WAT of SC-O mice, but this effect was greater in HFD-O mice. In conclusion these changes exacerbated cytokine production in response to LPS and contributed to the reduced sensitivity of the CAP.Cholinergic anti-inflammatory pathway (CAP) prevents inflammatory cytokines production. The main was to evaluate the effect of maternal obesity on cholinergic pathway in the offspring. Female mice were subjected to either standard chow (SC) or high-fat di422192202FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTIFICO E TECNOLOGICOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTIFICO E TECNOLOGICO14/18165-9; 13/10706-8476643/2012-014/18165-9; 13/10706-8476643/2012-0Les cages généralement utilisées en expérimentation animale peuvent empêcher les rats d'adopter la plupart des formes naturelles de comportement locomoteur. Ces animaux ont tendance a` adopter des habitudes sédentaires. Dans cette étude, nous démontrons

Autophagy proteins are modulated in the liver and hypothalamus of the offspring of mice with diet-induced obesity

2011/51.205-6Nutritional excess during pregnancy and lactation has a negative impact on offspring phenotype. In adulthood, obesity and lipid overload represent factorsthat compromise autophagy, a process of lysosomal degradation. Despite knowledge of the impact of obe343041FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2011/51.205-62011/51.205-

Low Birth Weight Intensifies Changes in Markers of Hepatocarcinogenesis Induced by Fructose Consumption in Rats.

Peer reviewed: TrueFunder: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Intrauterine growth restriction (IUGR) due to fetal exposure to glucocorticoid excess results in metabolic inflexibility and hepatic steatosis upon nutritional stress during adulthood. We previously demonstrated that rats born to dexamethasone (DEX)-treated mothers developed hepatic steatosis when exposed to 10% fructose solution during adult life. Persistent triacylglyceride (TAG) accumulation in the liver, in turn, is a feature of non-alcoholic fatty liver disease (NAFLD), which serves as a risk factor for non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). In the present study, we demonstrate that the combination of IUGR and fructose treatment during adulthood also results in increased hepatic myeloperoxidase (MPO) activity, AKT phosphorylation and serum aspartate transaminase. Growth-restricted rats also presented reduced hepatic TRIB3 and GADD45a after fructose treatment. Other markers of cell proliferation, such as Cyclin D, PCNA, Hgf and Hspa4/Hsp70 expression and the number of Ki-67 positive cells, were all increased in the liver of growth- restricted rats treated with fructose. On the other hand, the combination of IUGR and fructose treatment during adult life reduced the levels of IGF-1. In conclusion, our data indicate that after exposure to fructose, adult rats subjected to dexamethasone-induced IUGR display exacerbated molecular changes in markers of NASH and HCC

Inflammatory and insulin signalling proteins in hypothalamus at d28.

<p>Western blot analysis of p-JNK (A), p-IKK (B), PTP1B (C), p-IRS1 (D), p-AKT (E), and p-STAT3 (F) in the hypothalamus at d28. For control of gel loading, membranes were reblotted with β-actin or total proteins. Data are means ± SEM (n = 3–8). T test analysis was used. In all blots, at least three different litters were considered. Different letters indicate significant differences at p<0.05.</p