Orientador: Licio Augusto Velloso / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-27T14:54:51Z (GMT). No. of bitstreams: 1 Dragano_NathaliaRomanelliVicente_D.pdf: 1805761 bytes, checksum: fe734ef29b5f10bfd7ff5e19b729fec0 (MD5) Previous issue date: 2015 / Resumo: A recente caracterização da atividade anti-inflamatória atípica exercida pelo receptor de ácidos graxos poli-insaturados de cadeia longa GPR120 tem despertado grande interesse sobre esta classe de receptores, como potenciais alvos para o tratamento da obesidade e distúrbios metabólicos relacionados. Até o momento, a maioria dos estudos realizados tem explorado principalmente os benefícios metabólicos potenciais de uma ativação sistêmica dos receptores GPR120 e GPR40; no entanto, estudos recentes demonstraram que o hipotálamo é afetado logo durante as fases iniciais do desenvolvimento da obesidade desempenhando um papel crucial na patogênese desta doença e de suas comorbidades. Neste trabalho, nós avaliamos a expressão e potenciais ações terapêuticas do GPR120 e GPR40 no hipotálamo de camundongos obesos. Nós observamos que ambos os receptores são expressos no hipotálamo, sendo o GPR120 principalmente presente em células da microglia e o GRP40 preferencialmente expresso em neurônios orexigênicos NPY. Após o tratamento intracerebroventricular com GW9508, um agonista não específico destes receptores, foi observado uma redução da eficiência energética e da expressão de genes inflamatórios no hipotálamo de camundongos obesos. O silenciamento hipotalâmico do GPR120, por meio de lentivírus, aboliu o efeito anti-inflamatório induzido por GW9508 e resultou em maior eficiência energética nos animais tratados. Entretanto, o tratamento intracerebroventricular com os agonistas específicos do GPR120 e GPR40, TUG1197 e TUG905, respectivamente, resultaram em efeitos mais moderados sobre a homeostase energética e inflamação induzida por HFD do que aqueles observados com o GW9508. O TUG1197 atuou reduzindo a inflamação induzida por HFD por meio da redução da expressão das citocinas pró-inflamatórias, TNF? e IL1?, e aumento da expressão das interleucinas anti-inflamatórias, IL10 e IL6. O TUG905 promoveu a redução do peso corporal e da expressão do neuropeptídeo anorexigênico POMC. Em conclusão nosso trabalho demosntrou que os receptores GPR120 e GPR40 de forma coordenada no hipotálamo de roedores reduzindo a eficiência energética e regulando a inflamação associada à obesidade. A ativação de ambos os receptores no hipotálamo levou a melhores desfechos metabólicos se comparada com a ativação de qualquer um dos receptores isoladamente / Abstract: The recent characterization of the atypical anti-inflammatory activity exerted by the polyunsaturated fatty acid receptor GPR120 has raised the interest on this class of receptors as potential targets for the treatment of obesity and related disorders. Most studies performed to date have explored the potential metabolic benefits of a systemic activation of GPR120 and GPR40; however, it is currently known that the hypothalamus is affected during the early stages of obesity and plays an important role in the pathogenesis of this disease and its comorbidities. Here, we evaluate the expression and potential therapeutic actions of hypothalamic GPR120 and GPR40 in experimental diet-induced obesity. We show that both receptors are expressed in the hypothalamus; GPR120 is mostly present in microglia, whereas GRP40 is preferentially expressed in NPY neurons. Upon intracerebroventricular treatment, GW9508, a non-specific agonist of both receptors, reduced energy efficiency and the expression of inflammatory genes in the hypothalamus. Reducing GPR120 hypothalamic expression using a lentivirus approach resulted in the loss of the anti-inflammatory effect of GW9508 and increased energy efficiency. The intracerebroventricular treatment with GPR120 and GPR40 specific agonists, TUG1197 and TUG905, respectively, resulted in milder effects than those produced by GW9508. TUG1197 acted predominantly controlling inflammation reducing the expression of TNF? and IL1? while increasing IL10 and IL6. Conversely, TUG905 acted reducing body mass and increasing the expression of POMC. We conclude that GPR120 and GPR40 act in concert in the hypothalamus to reduce energy efficiency and regulate the inflammation associated with obesity. The combined activation of both receptors in the hypothalamus results in better metabolic outcomes than the isolated activation of either alone / Doutorado / Fisiopatologia Médica / Doutora em Ciências

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21340号 / 医博第4398号 / 新制||医||1031(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 妹尾 浩, 教授 岩井 一宏, 教授 横出 正之 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22827号 / 医博第4666号 / 新制||医||1047(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 井上 治久, 教授 髙橋 良輔, 教授 Shohab YOUSSEFIAN / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

In my study thesis Effect of eikosapentaenoic and dokosahexaenoic acids on expression of selected genes, which participate in modulation of inflammatory reaction at model organism I evaluated expression of genes GPR120, PPARgamma, LBP, ICAM, Il-4, Il-10, Il-1beta and TGF-beta. As model organism was selected pig. The pigs were divided in two groups, the control group was fed with 2,5% addition of palmic oil (P), the second group was fed with 2,5% addition of fish oil. After 70 days long fattening was each group divided into halfs. One half of each group was stimulated LPS (P+ and R+). Liver and adipose tissue were collected, mRNA was isolated (Rneasa Mini Kit), after reverse transcription the expression was measured by quantitative RT-PCR. By almost every genes there was increase in expression after LPS stimulation in groups P+ and R+. between P- and R- the expression was same or a little bit higher in R-. R+ was allways bigger than R-. The hypothesis were that EPA and DHA should reduce expression in inflammation. This hypothesis was not proofed. I recomend further studies with the same model organism to refill the results.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第18695号 / 農博第2092号 / 新制||農||1029(附属図書館) / 学位論文||H27||N4889(農学部図書室) / 31628 / 京都大学大学院農学研究科食品生物科学専攻 / (主査)教授 伏木 亨, 教授 河田 照雄, 教授 安達 修二 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19604号 / 医博第4111号 / 新制||医||1014(附属図書館) / 32640 / 京都大学大学院医学研究科医学専攻 / (主査)教授 妹尾 浩, 教授 松田 道行, 教授 川口 義弥 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DGAM

Orientador: Dennys Esper Corrêa Cintra / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Aplicadas / Made available in DSpace on 2018-08-25T14:56:46Z (GMT). No. of bitstreams: 1 Lazari_VanessadeOliveira_M.pdf: 4172597 bytes, checksum: 62cbaf2cfa729f497e7b3cc4cdf3e7a1 (MD5) Previous issue date: 2013 / Resumo: A obesidade é uma doença metabólica absolutamente fora de controle. As condições primárias associadas são hipertensão arterial, dislipidemias, diversos tipos de câncer, resistência à insulina (RI) e principalmente o diabetes mellitus tipo 2 (DM2). Dentre as características secundárias, esteatose hepática e síndrome do ovário policístico figuram entre as principais. Tais doenças acometem número cada vez maior de indivíduos e sobrecarregam sistemas públicos mundiais de saúde. A inflamação crônica e de baixo grau, designada "meta-inflamação", por acometer todo o organismo, parece ser o pano de fundo principal do processo obesogênico, resultando no descontrole da ingestão alimentar e na predisposição ao surgimento da RI podendo progredir para o DM2. Diante da ineficiência de terapêuticas farmacológicas atuais para o controle da obesidade, o investimento em estratégias anti-inflamatórias tem sido incentivado. Nesse intuito, ácidos graxos insaturados (AGIs), especialmente os ômega-3 (?3) e ômega-9 (?9) têm-se mostrado eficientes em atenuar a inflamação por meio da ativação do receptor GPR120, e em recuperar a sensibilidade à insulina em diversos tecidos. O objetivo deste trabalho foi avaliar as ações e mecanismos anti-inflamatórios dos AGIs ?-linolênico (?3) e oléico (?9), presentes em elevadas concentrações no óleo de semente de linhaça e oliva respectivamente, em tecidos periféricos metabolicamente ativos de animais obesos e diabéticos induzidos por dieta rica em gordura (HF), e suas repercussões na regulação glicêmica do organismo. O tratamento via dieta HF com substituição em 10% da fração saturada pelos respectivos óleos, foi eficiente em restabelecer o controle da fome dos animais que consequentemente ganharam menos peso em relação aos seus controles obesos. Dados fisiológicos obtidos por meio de medições de glicemia de jejum e dos testes de tolerância à insulina (ITT) e à glicose (GTT) evidenciaram melhoras na homeostase glicêmica, confirmado em nível molecular pelo aumento da atividade das principais proteínas envolvidas na via da insulina como o IR, IRS-1 e Akt. Possivelmente esses resultados deram-se em virtude da redução da expressão de peptídeos pró-inflamatórios como TNF-?, IL-6 e IL-1?, seguidos pelo aumento da expressão da IL-10, de caráter anti-inflamatório. No fígado, a melhora na sensibilidade à insulina, resultou em inibição da enzima GSK-3 propiciando o restabelecimento do controle hepático sobre o metabolismo da glicose. Análises por hematoxilina/eosina (HE) e imunohistoquímica evidenciaram menor acumulação lipídica e redução da infiltração de macrófagos M1 no parênquima hepático, resultando em melhora do fenótipo de esteatose hepática macrovesicular e recuperação funcional deste tecido. Houve diminuição da hipertrofia dos adipócitos, fato que contribuiu com a remissão da inflamação. Foi verificada a presença do receptor GPR120 nestas células, permitindo, portanto a entrada dos AGIs. A técnica de espectrometria de massa atestou a incorporação dos AGIs de interesse no fígado, sustentando a hipótese de que os efeitos benéficos encontrados ocorreram em decorrência das ações destes nutrientes. Esses achados permitem concluir que fontes alimentares dos AGs ?3 e ?9 modulam de forma potente a resposta inflamatória no organismo. Tais ações são controladas pelo GPR120, receptor comum a esses AGs, e parecem ocorrer de maneira tecido-específica com repercussões sistêmicas, positivas para o controle glicêmico / Abstract: Obesity is characterized as a metabolic disease absolutely out of control which nowadays reaches epidemic levels. The diseases associated to thiscomorbid as hypertension, dyslipidemia, several kinds of cancer, insulin resistance (IR), type 2 diabetes (DM2) and secondary conditions as hepatic steatosis and polycystic ovary syndrome affect a large number of people,burdening the public health system. The low grade chronic inflammation seems to be the main cause of the obesogenic processes, resulting in an uncontrolled food intake and the predisposition for insulin resistance (IR), which can lead to type II diabetes. Once there is no effective pharmacological therapy to treat obesity so far, the investment in anti-inflammatory approaches has been encouraged. In this regard, unsaturated fatty acid, mostly 3-omega (?3) and 9-omega (?9) have been shown effective in decreasing the inflammatory process by activating its common receptor, the GPR120, by recuperating the insulin sensibility in several tissues resulting in improvements in the systemic glucose homeostasis. The purpose of this study was to evaluate the potential antiinflammatory effects of the fatty acids ?-linolenic(?3) and oleic (?9), found in high concentrations in flaxseed and olive oil respectively on active metabolic tissue as liver, skeletal muscle and white adipose tissue of obese and diabetic animals induced by high fat diet, as well as its effects on insulin signaling and glycemic regulation. The high fat treatment with 10% replacement of the saturated portion by ?3 or ?9 oil was effective in restoring the control of food intake in the animals, which consequently gained less weight compared to the obese group. Physiological data from fasting glycemic measure, insulin and glucose tolerance test (ITT and GTT) have showed improvements in the glycemic homeostasis, which was molecularly confirmed by the increase in the activity of the main proteins involved in the insulin signaling pathway as the IR, IRS-1 and AKT. We believe that these results are due to the decrease in proinflammatory peptides expression as TNF-?, IL-6 and IL-1? and increase in the IL-10 expression, known by its anti-inflammatory effects. Improvements in insulin sensitivity in liver resulted in GSK-3 inhibition culminating in the glucose hepatic metabolic reestablishing which contributes to restore the control of plasma glucose levels. Hematoxilin/Eosin (HE) and Immunohistochemistry analysis showed lower lipid accumulation and decreased macrophage infiltration in hepatic parenchyma corroborating the reducing in macrovesicular steatosis and functional recovering of this tissue. There was reduction in adipocyte size, which contributed to "meta-inflammation" remission. We could note the presence of GPR120 receptor in membrane of these cells probably allowing the ?3 and ?9 entry. Mass spectrometry technique attested high incorporation of these fatty acids in liver suggesting the beneficial effects found were due to these nutrients actions. These findings allow us to conclude that alimentary sources of ?3 and ?9 fattty acids modulate effectively the systemic inflammatory response, resulting in the insulin sensibility restoration and in the glucose tolerance improvement. Such nutrients have shown to be important therapeutic nutrigenomic tools against obesity and its associated comorbidities / Mestrado / Nutrição / Mestra em Ciências da Nutrição e do Esporte e Metabolismo

Certains mammifères, dont l’Homme, ont une forte attraction pour les lipides alimentaires. Pendant longtemps, il était admis que ces nutriments étaient détectés uniquement via leurs propriétés olfactives, texturales et post-ingestives. Cependant, l’existence d’une dimension gustative a été suggérée depuis. Dans ce contexte, notre Laboratoire a démontré que la glycoprotéine CD36 exerçait une fonction de lipido-récepteur gustatif impliquée à la fois dans la préférence spontanée pour les lipides alimentaires et la phase céphalique de la digestion induite par la présence de lipides au niveau oral chez la souris. Une autre protéine, GPR120, semble également jouer un rôle dans la détection gustative des lipides alimentaires. Ces travaux de thèse avaient donc pour objectif de comprendre les rôles respectifs de ces deux protéines au sein des bourgeons du goût chez la souris. En utilisant une combinaison d'approches biochimiques, physiologiques et comportementales, nous avons pu montrer que le CD36 lingual était régulé négativement par les lipides alimentaires lors d'une exposition court terme, contrairement au GPR120. Cette régulation pourrait se traduire par une désensibilisation au cours du repas du système de lipido-réception gustatif et donc jouer un rôle dans le rassasiement sensoriel spécifique. Nos résultats suggèrent également que l'activation de GPR120 par les acides gras à longues chaînes peut jouer un rôle dans la modulation de la sensibilité gustative aux saveurs sucrées, voire aux lipides eux-mêmes. Ce phénomène est dépendant de la sécrétion de l'hormone glucagon-like peptide-1 par la papille caliciforme, indépendamment du CD36 lingual. Nous avons également mis en évidence que l'exposition des souris à un régime obésogène chronique perturbe la détection oro-sensorielle des lipides, suggérant une hyposensibilité chez la souris obèse. On peut donc penser qu'une perturbation du système de perception gustative des graisses alimentaires pourrait induire des changements de préférence alimentaire, propice à l’installation d'une obésité. De plus amples investigations sont requises pour explorer le rôle du CD36 et/ou du GPR120 dans ce phénomène. L'aboutissement de telles recherches serait l'émergence de nouvelles voies de traitement de l’obésité et des maladies associées, par des approches pharmacologiques et/ou nutritionnelles ciblées. / Some mammals including humans display a spontaneous attraction for dietary fat. For a long time, it was thought that these nutrients were detected only by olfactory, textural and post-oral cues. However, recent data have indicated that the sense of taste could also contribute to this perception in laboratory rodents. In this context, it was demonstrated by our Laboratory that the glycoprotein CD36 is likely a lingual lipid sensor, involved both in preference for long-chain fatty acids and cephalic phase of digestion secondary to an oral lipid stimulation in mice. Another protein, GPR120, seems also to play a role in the gustatory detection of lipids. The aim of this Thesis was to understand the respective roles of these putative gustatory lipid sensors in mice. Using a combination of biochemical, physiological and behavioral approaches, we showed that the lingual CD36 is negatively regulated by dietary fat during a short term exposure (i.e. during a meal), in contrast to GPR120. This regulation is reminiscent of the receptor desensitization and might play a role in the sensory-specific satiety phenomenon. Our data also suggest that GPR120 activation by long-chain fatty acids plays a role in the modulation of sweet taste sensitivity, and likely of fat taste itself. This phenomenon is dependent from the secretion of the glucagon-like peptide-1 hormone by the circumvallate papillae, independently of lingual CD36. Finally, we showed that chronic exposure of mice to an obesogenic diet disturbs the oro-sensory perception of dietary lipids, suggesting that obese mice become hyposensitive to lipids. This alteration might induce changes in feeding preferences which could lead to obesity. More investigations are required to better understand the role of CD36 and/or GPR120 in this phenomenon. This new field of research might lead to new ways of investigations for the treatment of obesity and related diseases, by using novel pharmacological and/or nutritional approaches.

Des études chez le rongeur et l’Homme ont montré que la détection orosensorielle des lipides alimentaires implique une dimension gustative. Deux lipido récepteurs candidats sont présents dans les bourgeons du goût : CD36, dont l’implication dans le « goût du gras » a été démontrée par des études menées au laboratoire chez la souris, et GPR120. Les travaux de cette thèse ont montré que le GPR120 n’intervient pas directement dans cette détection, mais permettrait de moduler la sensibilité aux lipides. Ce système gustatif permet d’adapter les choix alimentaires aux besoins énergétiques tout en satisfaisant au plaisir de la consommation d’aliments palatables. Cependant, les personnes obèses surconsomment les aliments riches en énergie. S’il existe des perturbations centrales des mécanismes de récompense, nos résultats suggèrent que le système de détection périphérique des lipides est aussi altéré chez les souris obèses. Une diminution réversible de la sensibilité gustative pour les lipides est observée chez des souris rendues obèses par un régime riche en acides gras saturés, conséquence d’une dérégulation de la signalisation calcique CD36-dépendante dans les papilles gustatives. Pour déterminer l’origine de ces perturbations, le rôle de l’endotoxémie métabolique (due à une augmentation du LPS plasmatique provenant du microbiote intestinal) observée lors de l’obésité a été testé. Nous avons montré que, pris hors du contexte de l’obésité, l’endotoxémie bas-bruit est insuffisante pour altérer à elle-seule la détection orale des lipides. L’origine de cette altération est donc multifactorielle, impliquant probablement une combinaison de modifications hormonales et inflammatoires. / Dietary lipids are detected by the gustatory system in rodents and humans. Two candidate lipid-receptors are found in taste buds: CD36, which is involved in the fat taste as shown by studies conducted in our laboratory, and GPR120. Our results show that GPR120 is not directly involved in the gustatory detection of lipids in mice, but could rather be involved in the modulation of the sensitivity for fat. When this gustatory system works properly, food choices can meet the organism’s energy needs. Besides, the pleasure brought by the consumption of palatable foods is satisfied. However, obese people often overconsume energy-dense food. In the central nervous system, perturbations of the reward mechanisms have been observed, but our data show that the peripheral detection system is also altered in obese mice. A reversible decrease in the gustatory sensitivity for lipids has been found in diet-induced obese mice (diet rich in saturated fatty acids). This phenomenon is the consequence of a deficiency in the regulation of the CD36-dependant calcium signaling in gustatory papillae. To determine the origin of these perturbations, the role of obese-associated metabolic endotoxemia (characterized by an increase in plasma LPS coming from the intestinal microbiota) was investigated. We showed that low-grade endotoxemia, when studied outside of the context of obesity, is insufficient to trigger an alteration of the oral lipid detection. The origin of this alteration is therefore multifactorial, probably involving a combination of hormonal and inflammatory modifications.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第24880号 / 医博第5014号 / 新制||医||1068(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 長船 健二, 教授 江木 盛時, 教授 妹尾 浩 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM