Au cours des dernières années, de nombreuses études ont porté sur les relations entre le microbiote intestinal et l'obésité. Des groupes bactériens ont été incriminés et des hypothèses proposées mais les mécanismes liant microbiote et obésité restent en grande partie inconnus. Le microbiote intestinal est constitué de plusieurs centaines d’espèces et est spécifique de chaque individu. Récemment, il a été révélé l’implication potentielle de ce microbiote dans le développement de l’obésité. Lors d’une étude précédente, notre équipe a ainsi montré que les souris sans germes (dites « axéniques ») sont résistantes à l’obésité et aux désordres métaboliques induits par un régime hypercalorique (Roy et al. 2012). Par ailleurs, nous avons observé que certaines souris conventionnelles soumises à un tel régime développent une obésité et une insulinorésistance importantes (phénotype dit répondeur) tandis que d'autres restent minces et tolérantes au glucose (phénotype dit non répondeur) indépendamment de la quantité de nourriture consommée. Les mécanismes expliquant cette hétérogénéité de phénotype sont actuellement peu connus. Ce projet de thèse vise donc à élucider les mécanismes liant le microbiote intestinal au développement de l'obésité et des désordres associés en partant du principe que l'hétérogénéité (en termes de composition et de fonctions) du microbiote intestinal, spécifique de chaque individu (aussi bien chez l'homme que chez le rongeur), joue un rôle dans ces réponses différentes à un même régime hypercalorique. Pour ce faire, des souris conventionnelles ont reçu un régime contrôle ou un régime hyperlipidique pendant 12 semaines. A l’issu de ce régime, des souris ont été sélectionnées en tant que répondeuses ou non-répondeuses Une approche par métagénomique à partir d’échantillons fécaux prélevés avant et après régime hyperlipidique et issus des souris répondeuses et non-répondeuses ont été analysés. Les profils de microbiotes avant et après régime hyperlipidique ont été comparés afin de déterminer les effets de ce régime sur le microbiote intestinal. De plus, le profil du microbiote des souris répondeuses a été comparé à celui des souris non-répondeuses afin de détecter des espèces bactériennes, des gènes ou des voies métaboliques sous- ou sur-représentés dans les deux phénotypes. Par ailleurs, des échantillons de fèces, d’urine et de plasma prélevés avant et après le régime hyperlipidique ont été analysés par métabolomique et nous avons ainsi analysé les fluctuations métaboliques induites par l’effet du régime. L’ensemble de ces travaux a eu pour but d’identifier s’il existe un microbiote de prédisposition au développement ou à la résistance du développement d’une obésité induite. Au cours de ce projet de thèse nous avons utilisé l’approche de métagénomique à partir d’échantillon fécaux afin d’identifier les profils du microbiote intestinal des souris NR et des souris R avant et après régime hyperlipidique en vue de mesurer l’impact de ce type de régime sur le microbiote, et surtout de détecter les espèces bactériennes, les gènes ou les voies métaboliques potentiellement sous- ou sur-représentés dans les deux phénotypes. D’autre part, l’approche de métabolomique a été utilisée sur des échantillons d’urine, de fèces et de plasma avant et après régime afin de visualiser les fluctuations métaboliques induites par l’effet du régime d’une part et d’identifier les métabolites (biomarqueurs) associés à chacun des phénotypes d’autre part. / In recent years, numerous studies have examined the relationship between the gut microbiota and obesity. Bacterial groups have been incriminated but the mechanisms linking microbiota and obesity remain largely unknown. Intestinal microbiota consists of several hundred species and is specific for each individual. Recently, it was revealed that the potential contribution of microbiota in the development of obesity. In a previous study, our team has shown that mice without germs (called "germ-free") are resistant to obesity and metabolic disorders induced by a high calorie diet (Roy et al. 2012). Furthermore, we observed that certain conventional mice subjected to such a plan develop an important obesity and insulin resistance (responder phenotype) while others remain thin and glucose tolerant (non responder phenotype) regardless of the amount of food consumed. The mechanisms explaining this phenotype heterogeneity are currently poorly known. This PhD project aims to elucidate the mechanisms linking the intestinal microbiota in the development of obesity and disorders associated with the assumption that heterogeneity (in terms of composition and functions) of the intestinal microbiota, specific for each individual ( both in humans than in rodents), plays a role in these different responses to the same high-fat diet. To do this, conventional mice received control diet or a high fat diet for 12 weeks. At the end of this diet, mice were selected as a responder or non-responder A metagenomics approach from fecal samples taken before and after high fat diet and from responder and non-responder mice were analyzed. Microbiota profiles before and after fat diet were compared to determine the effects of this diet on the intestinal microbiota. In addition, the profile of the microbiota of responder mice was compared to that of non-responder mice in order to detect bacterial species, genes or pathways under- or over-represented in both phenotypes. Furthermore, samples of feces, urine and plasma collected before and after fat diet were analyzed by metabolomics and we have analyzed the metabolic changes induced by the effect of diet. All of this work has been aimed to identify if there is a predisposition microbiota to the development or the resistance of induced obesity. In this PhD project we used the metagenomic approach from fecal sample to identify the profiles of the intestinal microbiota in mice NR and R mice before and after fat diet in order to measure the impact of this type of diet on the microbiota, and especially to detect bacterial species, genes or metabolic pathways potentially under- or over-represented in both phenotypes. On the other hand, the approach of metabolomics has been used on samples of urine, feces and plasma before and after diet in order to visualize the metabolic changes induced by the effect of diet on the one hand and to identify metabolites (biomarkers) associated with each other hand phenotypes.

Orientadores: Jaime Amaya-Farfan, Fernanda de Pace / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-27T02:35:33Z (GMT). No. of bitstreams: 1 Monteiro_NaiceEleidianeSantana_M.pdf: 1835989 bytes, checksum: ec62cf398adabcdca614249ae2d4692b (MD5) Previous issue date: 2015 / Resumo: Caracterizada como um dos mais importantes problemas que a saúde pública enfrenta atualmente no Brasil e no mundo, a obesidade está associada a um quadro de inflamação subclínica, que predispõe à resistência à insulina e ao desenvolvimento de diabetes mellitus, além de representar fator etiológico para diversas outras doenças crônicas não transmissíveis. Sabendo da importância que os alimentos com propriedades bioativas podem ter no tratamento da obesidade é que a utilização das proteínas do soro do leite na dieta vem sendo alvo de diversos estudos, devido aos benefícios que podem trazer à saúde humana. Dentre as propriedades da whey protein, citam-se a capacidade de regular a função imune, atuar como agente antimicrobiano, estimular a síntese de proteína muscular, suprimir o apetite e atuar na redução da gordura corporal, além das propriedades de aumentar a expressão gênica de proteínas anti-estresse (HSPs) e de ativar o transportador de glicose GLUT4, ambas descobertas pelo nosso grupo da FEA. O presente estudo teve como objetivo investigar o impacto que as proteínas do soro do leite, seja na sua forma íntegra ou na forma hidrolisada, possam exercer sobre a homeostase e a proteção do organismo de camundongos, contra os danos causados por uma dieta hiperlipídica. Para alcançar este objetivo, 34 camundongos C57BL/6 recém-desmamados foram divididos em quatro grupos de forma aleatória e alimentados com as seguintes dietas: grupo controle normolipídico (AIN93-G), grupo controle hiperlipídico (HFCAS), grupo hiperlipídico com substituição da caseína pela proteína do soro de leite concentrada (HFWPC), e grupo hiperlipídico, com adição da proteína do soro do leite hidrolisada (HFWPH), por 9 semanas. Os camundongos foram acompanhados quanto ao consumo alimentar e ganho de peso por meio de avaliação poderal em dias alternados, parâmetros bioquímicos e de endotoxemia avaliados por técnicas convencionais, biomarcadores inflamatórios analisados por western blot, além da avaliação do perfil histopatológico do fígado e do estudo metagenômico da microbiota intestinal. Os resultados demonstraram que alimentação com a formulação contendo whey protein, em ambas as formas, por 9 semanas, não diminuiu ganho de peso em comparação aos demais tratamentos, mas foi efetiva em reverter a disbiose causada pela dieta gordurosa, reduzir o processo inflamatório para níveis indistinguíveis do controle, em atenuar a infiltração gordurosa no tecido hepático, além de modular a microbiota intestinal. A caseína não mostrou tais propriedades / Abstract: Recognized as one of the most important issues currently facing public health in Brazil and in the world, obesity is associated to a subclinical inflammation framework, which predisposes to insulin resistance and the development of diabetes mellitus, in addition to representing an etiological factor for several other chronic diseases. Owing to the bioactive properties that the milk whey proteins may have on human health, such as the ability to regulate immune function, act as an antimicrobial agent, stimulate protein synthesis, suppress appetite thus helping to reduce body fat, besides the two newly discovered functions from our laboratory regarding the up-regulation of protective HSPs and activation of the glucose transporter 4 (GLUT4), it is thought that these proteins may also have anti-inflammatory action and, therefore, this study was designed to investigate the impact that whey protein, either in its normal form or in the hydrolyzed form, may exert on homeostasis protecting the mouse from the adverse effects of a high-fat diet. To accomplish this, 34 C57BL/6 male mice were randomly divided into four groups that received the following diets for 9 weeks: Normolipidic control (AIN93-G), Hyperlipidic control (HFCAS), Hyperlipidic with whey protein concentrate instead of casein (HFWPC) and a Hyperlipidic group with hydrolyzed whey protein (HFWPH) as the only source of protein. Diet intake and weight gain were monitored and recorded every other day. Biochemical parameters and endotoxemia were evaluated using commercial kits, and inflammatory biomarkers were analyzed by western blotting, besides the evaluation of histopathological liver profile and metagenomic study of the intestinal microbiota. The results showed that feeding the formulation contendowheyprotein, in both its forms, for 9 weeks, not decreased weight gain compared to the other treatments, but was effective in reversing dysbiosis caused by high-fat diet, reduce inflammation to levels indistinguishable control and mitigate the fatty infiltration of the casein hepático, in addition to modulate the gut microbiota. Casein did not show such properties / Mestrado / Nutrição Experimental e de Alimentos / Mestra em Alimentos e Nutrição

O estudo avaliou os efeitos da fototerapia na cicatrização de úlceras cutâneas e sua relação com estresse oxidativo experimental em ratos. Foram selecionados 80 ratos Wistar que foram divididos aleatoriamente em dois grupos de acordo com a dieta, controle e hiperlipídica (50% gordura) durante 45 dias. Posteriormente, foram avaliadas as concentrações séricas de glicemia, colesterol total, triglicérides, HDL e adiponectina, além da avaliação clínica e histopatológica dos fígados dos animais. Os animais normolipídicos (NL) e hiperlipídicos (HL) foram distribuídos em grupos (n=20) conforme tratamento fototerápico, a saber: aparelho desligado (sham) e ligado (laser diodo, 660 nm, 2 J, 70 J/cm&sup2, 100 mW) sendo designados NLL-, HLL-, NLL+ e HLL+ respectivamente. Duas úlceras cutâneas foram realizadas com punch 15 mm e irradiadas 3 vezes por semana durante 14 dias. As úlceras foram fotografadas e analisadas por meio do programa ImageJ e calculado os índices de cicatrização das úlceras (ICUs) no 2º, 7° e 14° dias, além da dosagem de hidroxiprolina para avaliação da colagênese e avaliação histopatológica (HE) qualitativa do processo cicatricial. Foram coletadas biópsias das lesões nos dias de seguimento para dosagens de mieloperoxidase (MPO), malondialdeído (MDA), peróxidos de hidrogênio (FOX), glutationa reduzida (GSH) para avaliar o estresse oxidativo tecidual. Os resultados mostraram que após o período de dieta, os animais hiperlipídicos apesar de não apresentarem aumento na massa corporal, apresentaram aumento da glicemia (p=0,0001), colesterol total (p=0,0002), triglicérides (p=0,0001) e diminuição nos níveis de adiponectina (p=0,003) comparado ao grupo normolipídico. Além disso, apresentaram fígados com alterações clínicas e histopatológicas compatíveis com quadro de esteatose hepática. Em relação ao estudo da cicatrização, no 14º dia o grupo hiperlipídico apresentou atraso na reepitelização das úlceras (p=0,0001), diminuição da colagênese (p=0,0001), e na análise histopatológica qualitativa do 14º dia evidenciou pequena formação de epiderme, poucos fibroblastos e colágeno desorganizado em relação aos normolipídicos. Quanto ao estresse oxidativo, no 2º dia os animais hiperlipídicos apresentaram aumento nos níveis de MPO (p=0,0001), MDA no 2º e 14º dia (p=0,0003 e p=0,0001), FOX no 2º e 14º dia (p=0,01 respectivamente) e GSH no 2º dia (p=0,009) em relação aos normolipídicos. Em relação aos grupos irradiados, no 2º dia o grupo NLL+ apresentou maiores ICUs em relação ao HLL- (p<0,05) e o HLL+ apresentou maiores ICUs em relação aos controles NLL- e HLL- (p<0,05). No 14º dia o grupo NLL+ apresentou novamente maiores ICUs em relação ao HLL- e o grupo HLL+ apresentou maiores ICUs em comparação com o HLL- e NLL+ (p<0,05). Quanto à colagênese, no 7º dia o NLL+ apresentou aumento em relação aos demais grupos (p<0,05) e no 14º dia o NLL+ continuou apresentando aumento em comparação com os outros grupos e o HLL+ apresentou aumento em relação ao HLL- (p<0,05). Em relação ao estresse oxidativo, no 2º dia o grupo NLL+ apresentou aumento de MPO em relação aos grupos NLL- e HLL+ (p<0,05). No 7º dia o grupo NLL+ apresentou diminuição nos níveis de MPO em comparação ao NLL- e o HLL+ apresentou diminuição em relação aos controles NLL- e HLL- (p<0,05). As dosagens de MDA no 2º dia mostraram que o grupo HLL+ apresentou menores índices em comparação ao HLL- e no 14º dia o NLL+ apresentou diminuição nos índices em relação ao HLL- (p<0,05). De acordo com as dosagens de FOX no 2º dia o NLL+ mostrou níveis elevados em comparação com os grupos NLL- e HLL+, e o grupo HLL+ mostrou níveis mais baixos em relação ao HLL- e NLL+ (p<0,05). Quanto as dosagens de GSH no 7º dia o grupo HLL+ apresentou níveis elevados em comparação aos demais grupos (p<0,05). Os resultados evidenciaram a influência negativa da dieta hiperlipídica nas concentrações séricas de glicemia, perfil lipídico, adiponectina e no fígado. Essas alterações metabólicas causaram ambiente de estresse oxidativo e diminuição da colagênese e atraso da reepitelização das úlceras cutâneas. Quanto à fototerapia, o grupo hiperlipídico irradiado acelerou a cicatrização de úlceras cutâneas por meio da modulação do estresse oxidativo em comparação ao grupo hiperlipídico não irradiado e aos grupos normolipídicos, sugerindo que efeitos benéficos similares podem ser encontrados em pacientes com alterações metabólicas. / The study evaluated the effects of phototherapy in the healing of skin ulcers and their relationship with oxidative stress in experimental rats. Eighty Wistar rats were divided were selected randomly into two groups according to diet, control diet and hyperlipidemic diet (50% fat) for 45 days. Subsequently, serum glucose concentrations were evaluated, cholesterol, total triglycerides, HDL and adiponectin, in addition to clinical and histopathological evaluation of liver animals. The normolipidemic (NL) and hyperlipidemic (HL) animals were divided into groups (n = 20) as phototherapy, namely laser off (sham) and laser on (laser diode, 660 nm, 2 J, 70 J/cm&sup2, 100 mW) being designated NLL-, HLL-, NLL+ and HLL+, respectively. Two skin ulcers were performed with a punch 15 mm and irradiated three times per week for 14 days. The ulcers were photographed and analyzed using the ImageJ program and calculated the rates of healing of ulcers in the 2nd, 7th and 14th days, besides the determination of hydroxyproline to evaluate collagenesis and histopathological evaluation (HE) qualitative healing process. Biopsies was collected on days of followup for determination of myeloperoxidase (MPO), malondialdehyde (MDA), hydrogen peroxide (FOX), reduced glutathione (GSH) to assess tissue oxidative stress. The results showed that after the diet period, the hyperlipidemic animals spite of not showing an increase in body mass, showed increased blood glucose levels (p = 0.0001), total cholesterol (p = 0.0002), triglycerides (p = 0.0001) and a decrease in adiponectin levels (p = 0.003) compared to normolipidemic group. Furthermore, livers presented with clinical and histopathological changes consistent with acute hepatic steatosis. Regarding the study of healing, on the 14th day hyperlipidemic group showed delay in re-epithelialization of ulcers (p = 0.0001), decreased collagenesis (p = 0.0001) , and qualitative histopathological analysis of the 14th day showed a small neo epidermis, few fibroblasts and disorganized collagen in relation to normolipidemic rats. Regarding oxidative stress, on the 2nd day the hyperlipidemic animals showed increased levels of MPO (p = 0.0001), MDA in the 2nd and 14th day (p = 0.0003 and p = 0.0001), FOX in the 2nd and 14th day (p = 0.01, respectively) and GSH on day 2 (p = 0.009) compared to normolipidemic. Regarding the irradiated groups, on the 2nd day the NLL+ group had higher WHR compared to HLL- (p < 0.05) and HLL+ WHR were higher compared to NLL- and HLL- (p < 0.05) control groups. On the 14th day the NLL + group showed higher again ICUs compared to HLL and HLL-+ group had higher ICUs compared with HLL and NLL + (p <0.05). Regarding collagenesis, on the 7th day the NLL+ showed an increase compared to the other groups (p < 0.05), on day 14 the NLL+ continued to show an increase compared with the other groups and HLL+ showed an increase compared to HLL- (p < 0.05). In relation to oxidative stress, on the 2nd day the NLL+ group showed increased MPO in relation to HLL+ and NLL- (p <0.05) groups. On the 7th day the NLL+ group showed a decrease in MPO levels in compared to the NLL-, and HLL+ showed a decrease compared to NLL+ and HLL- (p < 0.05) controls. Dosages of MDA on day 2 showed that the HLL+ group had lower rates compared the HLL- and on day 14 the NLL+ showed a decrease in rates compared to HLL- (p < 0.05). Of according to FOX dosages on the 2nd day the NLL+ showed high levels compared with HLL+ and NLL- groups and HLL+ showed lower levels compared to the HLLand NLL+ (p < 0.05). As the dosage of GSH on the 7th day the HLL+ group showed high levels compared to other groups (p < 0.05). The results showed the negative influence of high-fat diet on serum glucose, lipid profile, adiponectin and liver. Those metabolic changes caused oxidative stress, decreased collagenesis and delay in reepithelialization of cutaneous wounds. As for phototherapy, the group hyperlipidic irradiated accelerated the healing of skin ulcers by modulating oxidative stress in comparison to hyperlipidemic non-irradiated group and normolipidemic groups, suggesting that similar beneficial effects can be found in patients with metabolic disorders.

O consumo de dieta hiperlipídica e consequente acúmulo de lipídios nos adipócitos é uma condição associada ao estresse oxidativo e à perpetuação de um quadro inflamatório de leve intensidade que leva ao desenvolvimento de doenças crônicas não transmissíveis. Uma vez que alguns componentes da dieta são reconhecidos como fortalecedores do sistema antioxidante exógeno dos organismos vivos, o objetivo deste trabalho foi investigar o efeito metabólico do extrato de sementes do gênero Passiflora sobre parâmetros bioquímicos, oxidativos e inflamatórios de camundongos submetidos a uma dieta hiperlipídica. Para tanto, inicialmente realizou-se um estudo de composição química (centesimal, teores de minerais e ácidos graxos) e de otimização da extração de sementes de P. edulis Flavicarpa para obtenção de maiores teores de compostos polifenólicos com expressiva capacidade antioxidante in vitro, segundo os parâmetros de processo tempo, temperatura e concentração de etanol. Determinada a condição ideal de extração, esta foi empregada para as demais espécies em estudo P. alata BRS Mel do Cerrado e BRS Doce Mel, P. tenuifila BRS Vita e P. edulis BRS Sol do Cerrado e BRS Gigante Amarelo e P. setacea BRS Pérola do Cerrado e foi realizada o estudo de composição química. Os extratos etanólicos obtidos possuíram interessante atividade antioxidante, com destaque para a espécie P. setacea. O composto piceatanol foi o polifenol majoritário (0,41-10,28 g/ 100 g de semente em base seca) nas sementes de Passifloras analisadas, com exceção para a amostra P. setácea BRS Vita, cujo composto de íon molecular m/z 747,2 não foi identificado. As sementes ainda apresentaram alto teor de óleo, com o ácido linoleico em sua composição, e proteína. Os extratos das sementes de P. setacea BRS Pérola do Cerrado e Passiflora edulis Flavicarpa, nas concentrações de 500 e 1000 mg/Kg de ração foram utilizados para a realização do ensaio biológico. O consumo dos extratos, dependendo da dose, apresentou efeitos biológicos importantes, tais como a diminuição das concentrações séricas de colesterol, glicose, insulina e leptina a níveis aproximados ao determinado para os animais em consumo de dieta normolipídica. Adicionalmente, verificou-se a atenuação do estresse oxidativo hepático, por elevação da atividade enzimática das enzimas catalase e glutationa peroxidase e diminuição da lipoperoxidação, e do processo inflamatório, por redução da concentração tecidual das citocinas IL-6 e MCP-1. Os resultados obtidos neste estudo contribuem para o conhecimento a respeito das passifloras brasileiras e na potencial utilização das sementes, consideradas subprodutos, na contribuição da manutenção da saúde. / The consumption of a high fat diet and consequent excessive lipid accumulation in adipocytes is a condition associated with oxidative stress and perpetuation of a mild inflammatory condition that leads to the development of chronic diseases. Since some dietary components are recognized as empowering exogenous antioxidant system defenses of living organisms, the aim of this study is to investigate the metabolic effects of passion fruit seeds that possess a high content of bioactive compounds and high antioxidant capacity in vitro on oxidative stress and inflammatory conditions in mice subjected to a high fat diet. For this purpose, initially were performed a chemical composition study (proximate, minerals and fatty acids content) and extraction optimization of P. edulis Flavicarpa seed for obtaining higher levels of polyphenolic compounds with expressive antioxidant capacity in vitro through process parameters such as time temperature and ethanol concentration. Once the optimum condition of extraction was determined, it was applied to others studied species P. alata BRS Mel do Cerrado e BRS Doce Mel, P. tenuifila BRS Vita e P. edulis BRS Sol do Cerrado e BRS Gigante Amarelo e P. setacea BRS Pérola do Cerrado and the chemical composition study was carried out. The obtained ethanolic extracts had high antioxidant activity, particularly the species P. setacea. The piceatannol compound was the major polyphenol (0.41 to 10.28 g / 100 g seed in dry basis) in the analyzed Passiflora, except for the sample P. setacea BRS Vita, which molecular íon m/z 747.2 was not identified. The seeds also showed high content of oil, with linoleic acid in its composition, and protein. P. setacea BRS Pérola do Cerrado and Passiflora edulis Flavicarpa seed extracts at concentrations of 500 and 1000 mg / kg of feed were used to carry out the biological assay. The consumption of the extracts, depending on the concentration, exhibited significant biological effects, such as the reduction of serum cholesterol, glucose, insulin and leptin levels to those one observed in animals with normolipidic diet consumption. Additionally, hepatic oxidative stress was attenuated by elevating enzymatic activity of catalase and glutathione peroxidase and decrease in the lipid peroxidation and inflammation by reducing the tissue concentrations of IL-6 and MCP-1 cytokines. The results obtained in this study contributes to the knowledge of the Brazilian passiflora and potential use of the seeds, considered a by-products, in health maintenance.

No description available.

Liver mitochondria play a crucial role in intermediary metabolism and main metabolic pathways. We evaluated the pharmacological effect on liver mitochondria in vitro using two novel anticancer drugs: 3-bromopyruvate and α-tocopheryl succinate. Metabolic influence on liver mitochondria was performed in vivo by high fat and high cholesterol diet. Toxicity of both drugs was evaluated in cell cultures of hepatocytes isolated from rat and mouse liver. The effect of anticancer drugs on liver mitochondrial functions in vitro was studied on suspensions of isolated liver mitochondria, tissue homogenate and permeabilized hepatocytes. Mitochondrial respiration was measured using high-resolution respirometry. 3-bromopyruvate caused morphological and functional damage of primary rat and mouse hepatocytes in cell cultures; this toxic effect was accompanied by an increase of reactive oxygen species production and mitochondrial dysfunction. 3-bromopyruvate decreased the oxygen consumption of mitochondria energized by substrates for complex I and complex II. α-Tocopheryl succinate caused a decrease of succinate-dependent respiration in all experimental models both in coupled and in uncoupled states. The most pronounced effect of α-tocopheryl succinate was apparent in isolated mitochondria and the least pronounced...

Dietary supplementation of nicotinamide adenine dinucleotide (NAD+) precursors has been suggested as a treatment for non-alcoholic fatty liver disease and obesity. In the liver, NAD+ is primarily generated by nicotinamide phosphoribosyltransferase (NAMPT), and hepatic levels of NAMPT and NAD+ have been reported to be dependent on age and body composition. The aim of the present study was to identify time course-dependent changes in hepatic NAD content and NAD+ salvage capacity in mice challenged with a high-fat diet (HFD). We fed 7-week-old C57BL/6JBomTac male mice either regular chow or a 60% HFD for 6, 12, 24, and 48 weeks, and we evaluated time course-dependent changes in whole body metabolism, liver steatosis, and abundance of hepatic NAD-associated metabolites and enzymes. Mice fed a 60% HFD rapidly accumulated fat and hepatic triglycerides with associated changes in respiratory exchange ratio (RER) and a disruption of the circadian feeding pattern. The HFD did not alter hepatic NAD+ levels, but caused a decrease in NADP+ and NADPH levels. Decreased NADP+ content was not accompanied by alterations in NAD kinase (NADK) abundance in HFD-fed mice, but NADK levels increased with age regardless of diet. NAMPT protein abundance did not change with age or diet. HFD consumption caused a severe decrease in protein lysine malonylation after six weeks, which persisted throughout the experiment. This decrease was not associated with changes in SIRT5 abundance. In conclusion, hepatic NAD+ salvage capacity is resistant to long-term HFD feeding, and hepatic lipid accumulation does not compromise the hepatic NAD+ pool in HFD-challenged C57BL/6JBomTac male mice.

We have recently demonstrated that C57BL/6NTac and C57BL/6JRj substrains are significantly different in their response to high-fat diet-induced obesity (DIO). The C57BL/6JRj substrain seems to be protected from DIO and genetic differences between C57BL/6J and C57BL/6N substrains at 11 single nucleotide polymorphism (SNP) loci have been identified. To define genetic variants as well as differences in parameters of glucose homeostasis and insulin sensitivity between C57BL/6NTac and C57BL/6JRj substrains that may explain the different response to DIO, we analyzed 208 first backcross (BC1) hybrids of C57BL/6NTac and C57BL/6JRj [(C57BL/6NTac × C57BL/6JRj)F1 × C57BL/6NTac] mice. Body weight, epigonadal and subcutaneous fat mass, circulating leptin, as well as parameters of glucose metabolism were measured after 10 wk of high-fat diet (HFD). Genetic profiling of BC1 hybrids were performed using TaqMan SNP genotyping assays. Furthermore, to assess whether SNP polymorphisms could affect mRNA level, we carried out gene expression analysis in murine liver samples. Human subcutaneous adipose tissue was used to verify murine data of SNAP29. We identified four sex-specific variants that are associated with the extent of HFD-induced weight gain and fat depot mass. BC1 hybrids carrying the combination of risk or beneficial alleles exhibit the phenotypical extremes of the parental strains. Murine and human SC expression analysis revealed Snap29 as strongest candidate. Our data indicate an important role of these loci in responsiveness to HFD-induced obesity and suggest genes of the synaptic vesicle release system such as Snap29 being involved in the regulation of high-fat DIO.

Evidence suggests that obesity adversely affects brain function. High body mass index, hypertension, dyslipidemia, insulin resistance, and diabetes are risk factors for increasing cognitive decline. Tart cherries (Prunus Cerasus L.) are rich in anthocyanins and components that modify lipid metabolism. This study evaluated the effects of tart cherries on the brain in diet-induced obese (DIO) rats. DIO rats were fed with a high-fat diet alone or in association with a tart cherry seeds powder (DS) and juice (DJS). DIO rats were compared to rats fed with a standard diet (CHOW). Food intake, body weight, fasting glycemia, insulin, cholesterol, and triglycerides were measured. Immunochemical and immunohistochemical techniques were performed. Results showed that body weight did not differ among the groups. Blood pressure and glycemia were decreased in both DS and DJS groups when compared to DIO rats. Immunochemical and immunohistochemical techniques demonstrated that in supplemented DIO rats, the glial fibrillary acid protein expression and microglial activation were reduced in both the hippocampus and in the frontal cortex, while the neurofilament was increased. Tart cherry intake modified aquaporin 4 and endothelial inflammatory markers. These findings indicate the potential role of this nutritional supplement in preventing obesity-related risk factors, especially neuroinflammation.

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