Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A modulação do tecido adiposo marrom (TAM) e do tecido adiposo branco (TAB) está associada à prevenção ou redução do ganho de massa corporal. O óleo de peixe possui diversos efeitos benéficos que podem estar relacionados a esses tecidos. Dessa forma, objetivou-se avaliar os efeitos antiobesogênicos de diferentes dietas hiperlipídicas com óleo de peixe na termogênese do TAM e na lipogênese e beta-oxidação do TAB. Para isso, foram utilizados camundongos machos C57BL/6, com três meses de idade, que foram divididos em quatro grupos experimentais: um que recebeu dieta standard-chow (SC, 10% kcal de lipídios) e outros três que receberam dieta hiperlipídica (HL, 50% kcal de lipídios). Obtivemos os grupos HL com banha de porco (HL-B), HL com banha de porco mais óleo de peixe (HL-B+Px) e HL com óleo de peixe (HL-Px). As dietas foram administradas por um período de oito semanas, sendo que a ingestão alimentar foi avaliada diariamente e a massa corporal, semanalmente. Na última semana de experimento, realizou-se a calorimetria indireta e o teste oral de tolerância à glicose. No sacrifício, a glicemia foi aferida, o sangue foi puncionado para obtenção do plasma e o TAM interescapular e o TAB epididimário foram dissecados e armazenados. A leptina, os triglicerídeos e a insulina foram mensurados no plasma. O índice de adiposidade e o HOMA-IR foram calculados. O TAM e o TAB foram avaliados por microscopia confocal e de luz. Realizou-se RT-qPCR e Western blot para avaliação de marcadores termogênicos, da captação e oxidação de ácidos graxos e glicose e de PPAR no TAM, e para a avaliação da lipogênese e beta-oxidação e de PPAR no TAB. Com relação aos resultados, o grupo HL-B apresentou ganho de massa corporal e elevação da adiposidade, associado com hipertrofia dos adipócitos, hiperleptinemia, hipertrigliceridemia, intolerância à glicose e resistência à insulina, reproduzindo um quadro de obesidade e síndrome metabólica. Por outro lado, a ingestão de óleo de peixe nos dois grupos (HL-B+Px e HL-Px) foi capaz de reduzir o ganho de massa corporal e a adiposidade, sem alterar a ingestão alimentar. Essa ingestão também aumentou o gasto energético dos animais, regularizou a leptina e os triglicerídeos plasmáticos, bem como a tolerância à glicose e a resistência à insulina. Esses efeitos foram associados ao aumento de marcadores termogênicos no TAM, bem como da captação e oxidação de ácidos graxos e glicose e da expressão de PPAR nesse tecido. No TAB, houve redução de marcadores da lipogênese e aumento de marcadores da beta-oxidação, juntamente com elevação na expressão de PPAR. Em conclusão, nossos resultados mostram que a ingestão de óleo de peixe tem efeitos antiobesogênicos em camundongos através da modulação benéfica do TAM e do TAB e pode, portanto, representar uma terapia auxiliar alternativa contra a obesidade e suas comorbidades. / Brown adipose tissue (BAT) and white adipose tissue (WAT) modulation is associated with prevention or reduction of body mass gain. Fish oil has several beneficial effects which can be related to these tissues. Thus, we aimed to evaluate the anti-obesity effects of different high-fat diets with fish oil on BAT thermogenesis and WAT lipogenesis and beta-oxidation. For this, we used 3-mo-old C57BL/6 male mice that were divided into four groups: one that received a standard-chow diet (SC, 10% kcal of lipids) and three that received a high-fat diet (HF, 50% kcal of lipids). We obtained the HF with lard group (HF-L), the HF with lard plus fish oil group (HF-L+FO), and the HF with fish oil group (HF-FO). Diets were administrated for eight weeks, and food intake was evaluated daily and the body mass, weekly. At the end of the experiment, we performed indirect calorimetry and an oral glucose tolerance test. At sacrifice, glycemia was assessed, the blood was punctured to obtain plasma and interscapular BAT and epididymal WAT were dissected and stored. Plasmatic leptin, triglycerides and insulin were analyzed. Adiposity index and HOMA-IR were calculated. BAT and WAT were evaluated through confocal and light microscopy. RT-qPCR and Western blot were performed for analyses of thermogenic markers, fatty acids and glucose uptake and oxidation, and PPAR in BAT, and lipogenesis, beta-oxidation and PPAR in WAT. In relation to the results, the HF-L group presented elevated body mass gain and adiposity, associated with adipocyte hypertrophy, hyperleptinemia, hypertriglyceridemia, glucose intolerance and insulin resistance, displaying a condition that simulates obesity and metabolic syndrome. On the contrary, fish oil intake in both groups (HF-L+FO and HF-FO) was able to reduce body mass gain and adiposity, without affecting food intake. It also increased energy expenditure, normalized plasmatic leptin and triglycerides as well as glucose tolerance and insulin resistance. These effects were associated with increases in thermogenic markers, in uptake and oxidation of fatty acids and glucose and in PPAR expression in BAT. In WAT, lipogenesis was reduced and beta-oxidation and PPAR expression were increased. In conclusion, our results demonstrated that fish oil intake has anti-obesity effects in mice through beneficial modulation of BAT and WAT and can, therefore, represent an auxiliary alternative therapy against obesity and its comorbidities.

Afin de se maintenir au sein d'un environnement changeant, les individus doivent mettre en place une réponse adéquate. Il est connu que les animaux ont la capacité d'ajuster leur comportement à leur environnement. Cette plasticité comportementale, permet une réponse adaptée et relativement rapide aux variations de l'environnement, maximisant ainsi les chances de survie et de transmission des gènes. Elle met en jeu des processus cérébraux couteux en énergie rendant ces adaptations particulièrement sensibles à des changements alimentaires. Le but de cette thèse a été de mieux comprendre les facteurs qui contraignent ces réponses chez une espèce à laquelle s'applique une forte pression de sélection. Pour ce faire, nous avons étudié les réponses comportementales d'un primate malgache, le microcèbe gris (Microcebus murinus) soumis à des changements dans la quantité ou la qualité des ressources alimentaires disponibles. La première partie de ce travail s'est intéressée aux effets à court terme d'une restriction alimentaire sans malnutrition. Cette partie comprenait deux études. La première s'intéressant aux effets d'une restriction alimentaire à 60% sans malnutrition sur la plasticité comportementale innée via l’étude de l'horloge biologique. Les résultats de cette étude montrent une diminution de la capacité à se resynchroniser après un décalage horaire en lien avec la perte de poids. Ainsi, les individus perdant le plus de poids sont le moins à même de se resynchroniser sur les cycles lumineux après un décalage horaire de 6 heures. La seconde s'intéressait aux effets d'une restriction alimentaire de 40% sans malnutrition sur la plasticité comportementale acquise et montre une diminution de la capacité d'apprentissage des individus restreints après 19 jours de traitement alimentaire sans influence sur la mémoire à long terme. La moindre capacité d’apprentissage chez les individus en restriction calorique est corrélée à la perte de poids, les individus perdant le plus de poids étant les moins performants. Dans une seconde partie j’ai étudié l'effet de modifications qualitatives de l'alimentation à travers une supplémentation à long terme des individus en acides gras polyinsaturés n-3. Cette partie m’a permis de mettre en évidence une amélioration des performances d'apprentissage chez les individus supplémentés après 18 mois de traitement alimentaire accompagnée d'une diminution de l'anxiété et d'une augmentation de la neurogenèse adulte dans trois zones cérébrales.Ces travaux démontrent que les variations nutritionnelles, qu’elles soient quantitatives ou qualitatives sont capables d’influencer les différentes formes de plasticités comportementales et donc les grandes fonctions cérébrales et constituent ainsi un paramètre clé dans l’adaptation et la survie des individus. / In order to survive in a changing environment, individuals have to express an appropriate response. It is known that animals have the ability to adjust their behaviour to their environment. This behavioural plasticity allows a quick and adapted response to environmental variations, maximizing the individual'ssurvival and gene transmission. This plasticity relies on costly brain processes making these adaptations particularly dependent of food availability and maybe quality.This thesis project aimed at better understanding the constraints of these responses in a species under a strong selection pressure. To investigate this problematic, we studied the behavioural responses of a small Malagasy primate, the grey mouse lemur (Microcebus murinus), to both quantitative and qualitative changes in food resources. The first part of this work investigated the effect of a short-term caloric restriction without malnutrition over two studies. In the first one, we studied the effects of a 60% caloric restriction without malnutrition on innate behavioural plasticity via the study of the biological clock. The results show a decrease in the ability to resynchronize on a light/dark cycle following a time-shift. This difficulty to resynchronize was linked to body mass loss, the individuals loosing the more weight being the one unable to resynchronize after the 6-hours time shift. In the second study, we investigated the effect of a 40% caloric restriction without malnutrition on acquired behavioural plasticity. This study show a decrease in learning abilities of the restricted individuals after 19 days of dietary treatment and no influence on long term memory. This decrease in learning abilities was also linked with body mass loss, with the individuals loosing the more weight being the one with the worst success rate during this task. The second part focused on the effects of a qualitative variation in food supply via a long-term supplementation with n-3 polyunsaturated fatty acids. This part allowed us to show an increase in learning abilities associated with increased neurogenesis in three brain zones for supplemented animals after 18 month of treatment as well as a decrease of their anxiety level.This thesis work show that both quantitative and qualitative nutritional variations are able to influence different forms of behavioural plasticity and their cerebral basis and are of particular importance in the adaptation and survival of individuals.

SELECTED NUTRIENTS IN ETIOLOGY, PREVENTION AND TREATMENT OF OBESITY The 21st century pandemic of obesity is given by to the obesitogenic environment that helps to develop a positive energy balance, where the energy intake of an individual chronically exceeds his energy need, ie energy expenditure. There is an increase in body weight by increased or abnormal body fat accumulation in the body. Preventing the development of obesity is aimed at achieving energy balance, in the case of existing overweight or obesity, the treatment lies in inducing and maintaining a negative energy balance for some time, ie stimulating energy expenditure and reducing energy intake. Selected diet determines not only energy intake, but its individual nutrients can also partially affect energy expenditure and fat tissue physiology. The aim of this work is to describe and experimentally verify some nutrients that could help in weight reduction and physiological function of adipose tissue. Based on the literature data, 19 active substances or mixtures were considered. A mixture of ω-3 polyunsaturated fatty acids (ω-3 PUFA), eicosapentaenoic acid and docosahexaenoic acid, in a daily dose of 0.6 g, in the form of fish oil in a certified food supplement was chosen as the most suitable for the experiment. In a 12-week, three-arm, parallel...

SMOFLipid® is a commonly used fat emulsion for parenteral nutrition. We investigated how enrichment of SMOFLipid® with n-3 polyunsaturated fatty acids (PUFA) in a form of second fat emulsion, Omegaven® , changes fatty acid composition of total plasma phospholipids and erythrocyte phospholipids, cytokine concentrations in serum and in supernatant from in vitro whole blood culture stimulated with lipopolasaccharide (LPS) and we evaluated also changes in oxido- reductive balance. Eight patients on long-term home parenteral nutrition recieved both emulsions, SMOFLipid® (6 weeks) and SMOFLipid® +Omegaven® (4 weeks), one by one. We observed no significant differences in common laboratory and clinical parameters between these two types of diet. Enrichment of SMOFLipid® with Omegaven® led to an increase in eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) in total plasma phospholipids and there was also an increse in proportion of EPA in erythrocyte phospholipids, while proportion of DHA remained unchanged. These changes were in both phospholipids of plasma and erythrocyte compensated for a decrease in proportion of linoleic and arachidonic acid (n-6 PUFA). There were elevated IL-6 and TNF-α serum concentrations in patients after both diets. There was a decrease in IL-6 production by 36% with SMOFLipid®...

This thesis focuses on the effects of n-3 polyunsaturated fatty acids (n-3 PUFA) on development of non-alcoholic fatty liver disease (NAFLD) in experiment, on prevalence of this condition in patients with type 2 diabetes mellitus and metabolic syndrome and also on non-invasive diagnostics. The aim was to study the effect of n-3 PUFA on NAFLD development in an experimental model and based on analysis of a group of patients with type 2 diabetes and metabolic syndrome to assess the prevalence of this condition. Lastly we aimed to evaluate non-invasive diagnostic methods of liver fibrosis and NASH. We demonstrated beneficial effects of n-3 PUFA administration on NAFLD development in a C57/Bl6 mice high fat methionin-cholin defficient dietary model of NAFLD. n-3 PUFA administration led to biochemical improvement, decrease of lipid accumulation in the liver as well as improvement of histology. These effects are determined by complex modulation of lipid metabolism, mainly due to decrease in availability of fatty acids for triglyceride synthesis in the liver, changes of adipokine levels and amelioration of proinflammatory status in the liver. In a group of type 2 diabetics we found NAFLD prevalence of almost 80%, 14% of these patients had also signs of liver fibrosis or cirrhosis. Non-invasive methods...

This thesis focuses on the effects of n-3 polyunsaturated fatty acids (n-3 PUFA) on development of non-alcoholic fatty liver disease (NAFLD) in experiment, on prevalence of this condition in patients with type 2 diabetes mellitus and metabolic syndrome and also on non-invasive diagnostics. The aim was to study the effect of n-3 PUFA on NAFLD development in an experimental model and based on analysis of a group of patients with type 2 diabetes and metabolic syndrome to assess the prevalence of this condition. Lastly we aimed to evaluate non-invasive diagnostic methods of liver fibrosis and NASH. We demonstrated beneficial effects of n-3 PUFA administration on NAFLD development in a C57/Bl6 mice high fat methionin-cholin defficient dietary model of NAFLD. n-3 PUFA administration led to biochemical improvement, decrease of lipid accumulation in the liver as well as improvement of histology. These effects are determined by complex modulation of lipid metabolism, mainly due to decrease in availability of fatty acids for triglyceride synthesis in the liver, changes of adipokine levels and amelioration of proinflammatory status in the liver. In a group of type 2 diabetics we found NAFLD prevalence of almost 80%, 14% of these patients had also signs of liver fibrosis or cirrhosis. Non-invasive methods...