Type 2 diabetes mellitus is a metabolic disease characterised by defects in insulin secretion and insulin action and disturbances in carbohydrate, fat and protein metabolism. Hepatic insulin resistance contributes to hyperglycemia and also leads to disturbances in fat metabolism in type 2 diabetes. Psammomys obesus is a unique poly genie animal model of type 2 diabetes and obesity, ideally suited for studies examining physiological and genetic aspects of these diseases. To identify metabolic abnormalities potentially contributing to the obesity and diabetes phenotype in P. obesus, indirect calorimetry was used to characterise whole body energy expenditure and substrate utilisation. Lean-NGT, obese-IGT and obese-diabetic animals were examined in fed and fasted states and following 14 days of dietary energy restriction. Energy expenditure and fat oxidation were elevated in the obese-IGT and obese-diabetic groups in proportion to body weight. Glucose oxidation was not different between groups. Obese-diabetic P. obesus displayed elevated nocturnal blood glucose levels and fat oxidation. Following 14 days of dietary energy restriction, body weight was reduced and plasma insulin and blood glucose levels were normalised in all groups. Glucose oxidation was reduced and fat oxidation was increased. After 24 hours of fasting, plasma insulin and blood glucose levels were normalised in all groups. Energy expenditure and glucose oxidation were greatly reduced and fat oxidation was increased. Following either dietary energy restriction or fasting, energy expenditure, glucose oxidation and fat oxidation were not different between groups of P. obesus. Energy expenditure and whole body substrate utilisation in P. obesus was similar to that seen in humans. P. obesus responded normally to short term fasting and dietary energy restriction. Elevated nocturnal fat oxidation rates and plasma glucose levels in obese-diabetic P. obesus may be an important factor in the pathogenesis of obesity and type 2 diabetes in these animals. These studies have further validated P. obesus as an ideal animal model of type 2 diabetes and obesity. It was hypothesised that many genes in the liver of P. obesus involved in glucose and fat metabolism would be differentially expressed between lean-NGT and obese-diabetic animals. These genes may represent significant factors in the pathophysiology of type 2 diabetes. Two gene discovery experiments were conducted using suppression subtractive hybridisation (SSH) to enrich a cDNA library for differentially expressed genes. Experiment 1 used cDNA dot blots to screen 576 clones with cDNA derived from lean-NGT and obese-diabetic animals. 6 clones were identified as overexpressed in lean-NGT animals and 6 were overexpressed in obese-diabetic animals. These 12 clones were sequenced and SYBR-Green PCR was used to confirm differential gene expression. 4 genes were overexpressed (≥1.5 fold) in lean-NGT animals and 4 genes were overexpressed (≥1.5 fold) in obese-diabetic animals. To explore the physiological role of these genes, hepatic gene expression was examined in several physiological conditions. One gene, encoding thyroxine binding globulin (TBG), was confirmed as overexpressed in lean-NGT P. obesus with ad libitum access to food, relative to both obese-IGT and obese-diabetic animals. TBG expression decreased with fasting in all animals. Fasting TBG expression remained greater in lean-NGT animals than obese-IGT and obese-diabetic animals. TBG expression was not significantly affected by dietary energy restriction. TBG is involved in thyroid metabolism and is potentially involved in the regulation of energy expenditure. Fasting increased hepatic site 1 protease (SIP) expression in lean-NGT animals but was not significantly affected in obese-IGT and obese-diabetic animals. SIP expression was not significantly affected by dietary energy restriction. SIP is involved in the proteolytic processing of steroid response element binding proteins (SREBP). SREBPs are insulin responsive and are known to be involved in lipid metabolism. Gene expression studies found TBG and SIP were associated with obesity and diabetes. Future research will determine whether TBG and SIP are important in the pathogenesis of these diseases. Experiment 2 used SSH and cDNA microarray to screen 8064 clones. 223 clones were identified as overexpressed in lean-NGT P. obesus and 274 clones were overexpressed in obese-diabetic P. obesus (p ≤0.05). The 9 most significantly differentially expressed clones identified from the microarray screen were sequenced (p ≤0.01). 7 novel genes were identified as well as; sulfotransferase related protein and albumin. These 2 genes have not previously been associated with either type 2 diabetes or obesity. It is unclear why hepatic expression of these genes may differ between lean-NGT and obese-diabetic groups of P. obesus. Subsequent studies will explore the potential role of these novel and known genes in the pathophysiology of type 2 diabetes.

Submitted by (edna.saturno@ufrpe.br) on 2017-02-08T11:57:22Z No. of bitstreams: 1 Aprigio Marques de Sousa Neto.pdf: 1174109 bytes, checksum: b0262e812fcb006c6f76782c56519e12 (MD5) / Made available in DSpace on 2017-02-08T11:57:22Z (GMT). No. of bitstreams: 1 Aprigio Marques de Sousa Neto.pdf: 1174109 bytes, checksum: b0262e812fcb006c6f76782c56519e12 (MD5) Previous issue date: 2009-08-21 / In recent years, with the advent of global climate change, several studies have been developed in order to better understand the changes in the pelagic ocean environment and its effects on the main fishing resources explored in the three oceans. This present study was to evaluate the influences of environmental variability on the CPUE of bigeye tuna (Thunnus obesus) caught by Brazilian longline fleet in the period from 1978 to 2008. For this, we analyzed the data on settings, seasonality, fishing areas, effort, catch, time of immersion, number of hooks / Basket and fishing time, and some environmental data such as sea surface temperature (SST), the depth of the layer of mixture (PCM) and the rate of illumination of the moon (IIL). We performed a cluster analysis with the aim of categorizing the fishing effort associated with the catches of different species, then an analysis with generalized additive models (GAMS) in order to assess the relationship between these factors and the CPUE from big eye tuna. The results showed that as regards the variable latitude, the higher values of CPUE occurred around the equator. The case of the month on the CPUE index, we observed the highest values between October and April. Already the level of lunar illumination on the CPUE showed an increasing trend of the new moon to crescent. The SST showed a positive trend with higher values occurring between 26 and 30° C on CPUE. The effect of PCM, highest CPUEs occurred in areas where the thermocline is closer to the surface, between 40 and 70 m. In relation to the variable year, positive effects were only observed from 2000. These results are probably related to the reproductive activity of the specie, which is focused off the Brazilian coast for spawning. / Nos últimos anos, com o advento das mudanças climáticas globais, diversos estudos vêm sendo desenvolvidos no intuito de melhor compreender às variações do ambiente pelágico oceânico e seus efeitos sobre os principais recursos pesqueiros explorados nos três oceanos.O presente estudo teve como objetivo avaliar as influências da variabilidade ambiental sobre a CPUE da albacora bandolim (Thunnus obesus) capturada pela frota espinheleira brasileira no período de 1978 à 2008. Para isto, foram analisados os dados referentes aos lançamentos, sazonalidade, área de pesca, esforço, captura, tempo de imersão, número de anzóis/samburá e turno de pesca, e alguns dados ambientais, como a Temperatura da Superfície do Mar (TSM), a Profundidade da Camada de Mistura (PCM) e o Índice de Iluminação da Lua (IIL). Foi realizada uma análise de cluster com o objetivo de categorizar o esforço de pesca associandoo as capturas de diferentes espécies e, em seguida, uma análise com Modelos Aditivos Generalizados (GAMs) com o intuito de avaliar a relação entre estes fatores e a CPUE da albacora bandolim. Os resultados obtidos mostraram que no que se refere a variável latitude, os maiores valores de CPUE ocorreram no entorno do equador. Se tratando do índice mês sobre a CPUE, observou-se os maiores valores entre outubro e abril. Já o índice de iluminação lunar sobre a CPUE apresentou uma tendência crescente de lua nova para quarto-crescente. A TSM mostrou uma tendência positiva com maiores valores ocorrendo entre 26° e 30°C sobre a CPUE. No efeito da PCM, as CPUEs mais elevadas ocorreram em áreas onde a termoclina encontra-se mais próxima da superfície, entre 40 e 70 m. Em relação à variável ano, efeitos positivos só foram observados a partir de 2000. Estes resultados estão provavelmente associados à atividade reprodutiva da espécie, que se concentra ao largo da costa brasileira para desova.

Submitted by (edna.saturno@ufrpe.br) on 2017-02-16T14:56:11Z No. of bitstreams: 1 Marina Bezerra Figueiredo.pdf: 4281028 bytes, checksum: 769a66f89e02f5c1359899b741605fea (MD5) / Made available in DSpace on 2017-02-16T14:56:11Z (GMT). No. of bitstreams: 1 Marina Bezerra Figueiredo.pdf: 4281028 bytes, checksum: 769a66f89e02f5c1359899b741605fea (MD5) Previous issue date: 2007-02-23 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The study presents information about the reproductive characterísticas of bigeye tuna (Thunnus Obesus0. Were obtained 1082 samples of this species in the tropical Western Atlantic between July of 2004 and June of 2006. Were realized 873 (522 females and 351 males) histological cuts and observations of the present oocytes, analyzed with microscopy to determine the stage of gonadal development. For the females were determined 5 maturational stages: immature, initial and advanced maturation, spawning and resting. The fork length of the samples varied from 62 to 210 cm, and the sexual ratio was 1.46:1 (female:male). The females of bigeye tuna with high values of Gonadossomatic Index associated with advanced maturation were found during the first trimester, suggesting that this species has bigger reproductive activity in this period.The L for females was 92.8 cm and the males 82.1 cm. The estimate mean fecundity through hydrated oocytes counting for this species was 3,971,328 oocytes. / O presente estudo fez um levantamento sobre as características reprodutivas da albacora bandolim (Thunnus Obesus). Para isso, foi obtido um total de 1082 amostras de gônadas da presente espécie no Atlântico oeste Tropical entre julho de 2004 e junho de 2006. foram realizados 873 (522 fêmeas r 351 machos) cortes histológicos e observações dos ovócitos presentes, analisados microscopicamente para determinação das fases de desenvolvimento gonadal. Para as fêmeas foram determinados 5 estágios maturacionais: maturo, maturação inicial e avançada, desovado e em repouso. O comprimento zoológico dos exemplares variou de 62 210 cm, e a proporção sexual foi de 1,46 fêmeas para um macho. A relação entre o Índice Gonadal e exemplares em maturação avançada mostrou que a albacora bamdolim parece possuir maior atividade reprodutiva no primeiro trimestre do ano. O tamanho de primeira maturação sexual para as fêmeas foi de 92,8 cm, já para os machos esse valor foi menos, 82,1 cm. A fecundidade média estimada através da contagem de ovócitos hidratados para a presente espécie foi de 3.971.328,13 ovócitos.

Defects in fat metabolism are central to the aetiology and pathogenesis of obesity and type II diabetes. The liver plays a central role in these disease states via its regulation of glucose and fat metabolism. In addition, accumulation of fat within the liver has been associated with changes in key pathways of carbohydrate and fat metabolism. However a number of questions remain. It is hypothesised that fat accumulation within the liver is a primary defect in the aetiology and pathogenesis of obesity and type II diabetes. Fat accumulating in the liver is the result of changes in the gene expression of key enzymes and proteins involved with fat uptake, fat transport, fat oxidation, fat re-esterification or storage and export of fat from the liver and these changes are regulated by key lipid responsive transcription factors. To study these questions Psammomys obesus was utilised. This polygenic rodent model of obesity and type II diabetes develops obesity and diabetes in a similar pattern to susceptible human populations. In addition dietary and environmental changes to Psammomys obesus were employed to create different states of energy balance, which allowed the regulation of liver fat gene expression to be examined. These investigations include: 1) Measurement of fat accumulation and fatty acid binding proteins in lean, obese and diabetic Psammomys obesus. 2) Characterisation of hepatic lipid enzymes, transport protein and lipid responsive transcription factor gene expression in lean, obese and diabetic Paammomys obesus. 3) The effect of acute and chronic energy restriction on hepatic lipid metabolism in Psammomys obesus. 4) The effect of sucrose feeding on the development of obesity and type II diabetes in Psammomys obesus. 5) The effect of nicotine treatment in lean and obese Psammomys obesus, 6) The effect of high dose leptin administration on hepatic fat metabolism in Psammomys obesus. The results of these studies demonstrated that fat accumulation within the liver was not a primary defect in the aetiology and pathogenesis of obesity and type II diabetes. Fat accumulating in the liver was not the result of changes in the gene expression of key enzymes and proteins involved in hepatic fat metabolism. However changes in the mRNA level of the transcription factors PPAR∝ and SREBP-1C was associated with the development of diabetes and the gene expression of these two transcription factors was associated with changes in diabetic status.

Distribution of the novel peptide beacon in the hypothalamus of Sprague-Dawley rats was examined by immunohistochemical methods. Beacon-immunoreactive (irBC) neurons were found in the paraventricular, supraoptic, and accessory neurosecretory nuclei, and intensely labeled fibers in the median eminence and infundibulo-pituitary stalk. Scattered cells and/or fibers were noted in the suprachiasmatic nucleus, arcuate nucleus, retrochiasmatic area, lateral and medial preoptic area, as well as anterior and lateral hypothalamic area. The wide distribution of irBC in the hypothalamus of Sprague-Dawley rats suggests that the peptide may influence, in addition to a proposed role in feeding, a multitude of biological activities associated with the hypothalamic-pituitary axis.

La résistance à l’insuline et le diabète de type 2 (DT2) sont caractérisés par une hyperlipidémie. Le but de cette étude est de déterminer si le DT2 contribue au dérèglement du métabolisme du cholestérol au niveau du petit intestin et du foie du Psammomys obesus, un modèle animal nutritionnel d’induction de la résistance à l’insuline et du DT2. L’absorption intestinale du cholestérol est diminuée chez les animaux diabétiques. Cette diminution est associée à une baisse (i) de l’expression génique et protéique de NPC1-L1 qui joue un rôle primordial dans l’absorption du cholestérol au niveau des entérocytes; et (ii) de l’ARNm de l’ABCA1 responsable de l’efflux de cholestérol des cellules intestinales à l’apolipoprotéine A-I et aux HDLs. En ce qui a trait aux transporteurs SR-B1 et Annexin II, aucune différence n’a été observée au niveau intestinal. Toutefois, une diminution significative de l’expression génique de l’ABCG5, un intervenant majeur dans la sécrétion du cholestérol des entérocytes vers la lumière intestinale, est mesurée chez les animaux diabétiques. De plus, l’expression protéique est diminuée pour le PCSK9 et augmentée pour le LDLr au niveau du jéjunum, tandis que la quantité de protéine de l’enzyme HMG-CoA réductase est régulée à la baisse chez les Psammomys obesus diabétiques. Finalement, de tous les facteurs de transcription testés seule une augmentation de LXR et une diminution de PPAR/δ sont détectées au niveau de l’intestin. Au niveau hépatique, il y a (i) une augmentation de la masse protéique de NPC1-L1, SR-BI et Annexin II; (ii) une élévation l’ARNm de SR-BI; (iii) une diminution du contenu protéique de ABCG8 et de l’expression génique de l’ABCG5 et de l’ABCA1; et (iv) une élévation de l’ARNm de LXR et de PPAR/δ, tout comme une baisse de l’expression protéique de SREBP-2. Somme toute, nos résultats montrent que le développement du diabète de type 2 chez le Psammomys obesus entraîne un changement dans la machinerie intra-entérocytaire et hépatocytaire, qui mène à un dérèglement de l’homéostasie du cholestérol. / Insulin resistance and type 2 diabetes (T2D) are characterized by hyperlipidemia. The aim of the present study was to elucidate whether T2D contributes to abnormal cholesterol homeostasis in the small intestine and liver of Psammomys obesus, a model of nutritionally induced insulin resistance and type 2 diabetes. Diabetic animals exhibited a lower intestinal cholesterol uptake, which was associated with a decrease in (i) the gene and protein expression of NPC1L1, which plays a pivotal role in cholesterol incorporation in the enterocytes; and (ii) mRNA of ABCA1 that mediates cholesterol efflux from intestinal cells to apolipoprotein A-I and HDL. No changes were observed in the other intestinal transporters SR-BI and Annexin II. On the other hand, in diabetic animals, a significant mRNA decrease was noticed in ABCG5 responsible for the secretion of absorbed cholesterol back into the lumen. Furthermore, jejunal PCSk9 protein was diminished and LDLr was raised, along with a significant downregulation in jejunal HMG-CoA reductase in diabetic Psammomys obesus. Finally, among the transcription factors tested, only an increase in LXR and a decrease in PPAR/δ were detected in the intestine. In the liver, there was (i) an augmentation in the protein mass of NPC1L1, SR-BI and Annexin II; (ii) an upregulation of SR-BI mRNA; (iii) a fall in ABCG8 protein content, ABCG5 mRNA and ABCA1 mRNA; and (iv) an augmentation in LXR and PPAR/δ mRNA, as well as a drop in SREBP-2 protein. Overall, our findings show that the development of type 2 diabetes in Psammomys obesus modifies the whole intra-enterocyte and hepatocyte machinery, causing alterations in cholesterol homeostasis.

Le thon obèse (Thunnus obesus), le thon alabore (Thunnus albacares) et le listao (Katsuwonus pelamis) comptent parmi les espèces de thons les plus utilisées en conserve. Lors de la fabrication de conserves de thon, la substitution d'espèce et/ou le mélange de différentes espèces de thon sont interdits par la réglementation européenne. L'authentification des espèces de thon reste complexe à cause du degré de similitude élevé entre les espèces de thon, ou encore, lorsque les caractéristiques morphologiques externes sont éliminées au cours du filetage et lors de la mise en conserve. Par conséquent, des substitutions involontaires ou frauduleuses peuvent se produire. Dans cette étude, le marqueur mitochondrial du gène de la sous-unité 2 de la NADH déshydrogénase a été utilisé pour identifier le thon obèse et le gène de la sous-unité II de la cytochrome c oxydase a été utilisé pour identifier le thon albacore et le listao en utilisant la PCR en temps réel basée sur la technologie TaqMan. Deux méthodes différentes basées sur la qPCR ont été développées pour quantifier le pourcentage de chair de chaque espèce présente au sein d'une boîte de thon. La première a été basée sur la quantification absolue avec standard externe réalisée avec les deux marqueurs. La seconde a été basée sur la quantification relative avec standard externe avec le gène endogène de l'ARN 12S. Sur la base de ces résultats, nous pouvons conclure que notre méthode peut s'appliquer pour quantifier les deux espèces de thon albacore et obèse génétiquement très proches lorsqu'elles sont utilisées dans un mélange binaire en conserve. / Bigeye tuna (Thunnus obesus), yellowfin tuna (Thunnus albacares) and skipjack tuna (Katsuwonus pelanis) are among the most widely used tuna species for canning purposes. Not only substitution but also mixing of tuna species is prohibited by the European regulation for canned tuna products. However, it can be difficult to authenticate the tuna species, due to their high degree of similarity or even when the external morphological characteristics are removed due to filleting before canning. Consequently, involuntary or fraudulent substitutions may occur during the canning process. In this study, the mitochondrial marker from NADH dehydrogenase subunit 2 gene was used to identify bigeye tuna and the mitochondrial marker cytochrome c oxidase subunit II gene was used to identify yellowfin tuna and skipjack tuna, utilizing TaqMan qPCR methodology. Two different qPCR-based methods were developed to quantify the percentage of flesh of each species used for can processing. The first one was based on absolute quantification using standard curves realized with these two markers ; the second one was founded on relative quantification with the universal 12S rRNA gene as the endogenous gene. On the basis of our results, we conclude that our methodology could be applied to authenticate the two closely related tuna species (bigeye tuna and yellowfin tuna) when used in a binary mix in tuna cans.

La physiopathologie du diabète de type II se caractérise par de sévères anomaliesmétaboliques telles que l’hyperglycémie et les dyslipidémies contribuant au développementdes maladies cardiovasculaires. Une altération de l’activité de l’AMPK dans les tissus tels quele muscle squelettique et le foie est associée à ces désordres métaboliques alors que sonactivation pharmacologique permet de les rétablir. Toutefois, le complexe hétérotrimériqueαβγ tissu-spécifique de l’AMPK confère une régulation et des rôles distincts qui demeurentinexplorés dans l’intestin, un organe favorisant pourtant l’augmentation de l’absorption desnutriments en situation de diabète de type II. La présente étude démontre une prépondérancedu complexe α1β2γ1 de l’AMPK dans les cellules intestinales Caco-2 dont l’un des rôles de lasous-unité α1 est de réguler l’ACC, l’enzyme de synthèse des acides gras. Contrairement àl’AMPK exprimée dans le foie, elle ne régule pas l’HMG-CoA Réductase impliquée dans lasynthèse du cholestérol. L’activation de l’AMPK mime l’effet de l’insuline en réduisantl’absorption intestinale du glucose et des lipides alors que son altération en situationd’insulino-résistance (e.g : induite par le 4-HHE dans un modèle cellulaire Caco-2 ou induitepar la diète dans le modèle animal Psammomys obesus) favorise l’absorption du glucose etdes lipides, ce qui exacerberait l’hyperglycémie et la dyslipidémie postprandiale associées audiabète de type II. L’AMPK au niveau intestinal constitue donc une cible thérapeutiquepotentielle complémentaire pour la prévention et le traitement du diabète de type II. / Physiopathology of type II Diabetes is characterized by severe metabolic abnormalities suchas hyperglycemia and dyslipidemia also implicated in development of cardiovasculardiseases. Impaired AMPK activity in tissues such as skeletal muscle and liver is associatedwith these metabolic disorders whereas its pharmacologic activation is able to restore suchabnormalities. Nevertheless, tissue-specific heterotrimeric αβγ AMPK likely confers distinctroles and regulation that remain unexplored in intestine, an organ promoting enhancednutrients absorption in type II diabetes situation. This study demonstrates α1β2γ1 AMPKcomplex preponderance in intestinal Caco-2 cells whose α1 subunit role is to regulate ACCenzyme responsible of fatty acid synthesis. Unlike in the liver, AMPK doesn’t regulate HMGCoAreductase enzyme implicated in cholesterol synthesis. AMPK activation mimics insulineffect by reducing intestinal glucose and lipids absorption whereas its alteration in insulinresistancesituation (e.g.: induced by 4-HHE in Caco-2 cell model or in Psammomys obesusanimal model) enhances glucose and lipids absorption which could exacerbate postprandialhyperglycemia and dyslipidemia associated to type II diabetes. Thus, AMPK at the intestinallevel could be a potential therapeutic target in prevention and treatment of type II diabetes

La résistance à l’insuline et le diabète de type 2 (DT2) sont caractérisés par une hyperlipidémie. Le but de cette étude est de déterminer si le DT2 contribue au dérèglement du métabolisme du cholestérol au niveau du petit intestin et du foie du Psammomys obesus, un modèle animal nutritionnel d’induction de la résistance à l’insuline et du DT2. L’absorption intestinale du cholestérol est diminuée chez les animaux diabétiques. Cette diminution est associée à une baisse (i) de l’expression génique et protéique de NPC1-L1 qui joue un rôle primordial dans l’absorption du cholestérol au niveau des entérocytes; et (ii) de l’ARNm de l’ABCA1 responsable de l’efflux de cholestérol des cellules intestinales à l’apolipoprotéine A-I et aux HDLs. En ce qui a trait aux transporteurs SR-B1 et Annexin II, aucune différence n’a été observée au niveau intestinal. Toutefois, une diminution significative de l’expression génique de l’ABCG5, un intervenant majeur dans la sécrétion du cholestérol des entérocytes vers la lumière intestinale, est mesurée chez les animaux diabétiques. De plus, l’expression protéique est diminuée pour le PCSK9 et augmentée pour le LDLr au niveau du jéjunum, tandis que la quantité de protéine de l’enzyme HMG-CoA réductase est régulée à la baisse chez les Psammomys obesus diabétiques. Finalement, de tous les facteurs de transcription testés seule une augmentation de LXR et une diminution de PPAR/δ sont détectées au niveau de l’intestin. Au niveau hépatique, il y a (i) une augmentation de la masse protéique de NPC1-L1, SR-BI et Annexin II; (ii) une élévation l’ARNm de SR-BI; (iii) une diminution du contenu protéique de ABCG8 et de l’expression génique de l’ABCG5 et de l’ABCA1; et (iv) une élévation de l’ARNm de LXR et de PPAR/δ, tout comme une baisse de l’expression protéique de SREBP-2. Somme toute, nos résultats montrent que le développement du diabète de type 2 chez le Psammomys obesus entraîne un changement dans la machinerie intra-entérocytaire et hépatocytaire, qui mène à un dérèglement de l’homéostasie du cholestérol. / Insulin resistance and type 2 diabetes (T2D) are characterized by hyperlipidemia. The aim of the present study was to elucidate whether T2D contributes to abnormal cholesterol homeostasis in the small intestine and liver of Psammomys obesus, a model of nutritionally induced insulin resistance and type 2 diabetes. Diabetic animals exhibited a lower intestinal cholesterol uptake, which was associated with a decrease in (i) the gene and protein expression of NPC1L1, which plays a pivotal role in cholesterol incorporation in the enterocytes; and (ii) mRNA of ABCA1 that mediates cholesterol efflux from intestinal cells to apolipoprotein A-I and HDL. No changes were observed in the other intestinal transporters SR-BI and Annexin II. On the other hand, in diabetic animals, a significant mRNA decrease was noticed in ABCG5 responsible for the secretion of absorbed cholesterol back into the lumen. Furthermore, jejunal PCSk9 protein was diminished and LDLr was raised, along with a significant downregulation in jejunal HMG-CoA reductase in diabetic Psammomys obesus. Finally, among the transcription factors tested, only an increase in LXR and a decrease in PPAR/δ were detected in the intestine. In the liver, there was (i) an augmentation in the protein mass of NPC1L1, SR-BI and Annexin II; (ii) an upregulation of SR-BI mRNA; (iii) a fall in ABCG8 protein content, ABCG5 mRNA and ABCA1 mRNA; and (iv) an augmentation in LXR and PPAR/δ mRNA, as well as a drop in SREBP-2 protein. Overall, our findings show that the development of type 2 diabetes in Psammomys obesus modifies the whole intra-enterocyte and hepatocyte machinery, causing alterations in cholesterol homeostasis.

Nous avons étudié l’expression et la régulation de la lipogenèse dans les aortes et CMLV et déterminé si elle est modifiée par l’insulino-résistance et le diabète. Les rats Zucker obèses (ZO), diabétiques et Psammomys obesus accumulent plus de lipides dans leurs aortes que leurs contrôles. Cependant l’expression des gènes de la lipogenèse et ceux impliqués dans la captation des acides gras, n’est pas augmentée. Un résultat similaire a été retrouvé dans des pièces d’endartériectomies chez l’homme. Le milieu adipogénique, le glucose ou l’insuline seule stimulent modérément la lipogenèse uniquement dans les CML de Zucker contrôles, aucun effet n’a été observé dans les CML de ZO. Nous avons montré que les effets du TO901317 sur la lipogenèse dans les CMLV sont dus uniquement à l’activation du récepteur nucléaire LXRα, PXR n’a aucun effet. En conclusion, la lipogenèse n’est pas augmentée dans la paroi artérielle durant l’insulino-résistance et le diabète. / We investigated the expression and regulation of lipogenesis in aortas and VSMC and determined if it is modified during metabolic abnormalities. Zucker obese (ZO), diabetic (ZDF) rats, and the high fat diet fed Psammomys obesus accumulated more triglycerides in their aortas than control rats. However the expression of lipogenic genes, or of genes involved in fatty acids uptake, was not increased. Lipogenesis was not increased in human carotid endarterectomy of diabetic compared to non-diabetic patients. The adipogenic medium (ADM), glucose or insulin stimulated moderately lipogenesis but only in VSMC from control rats. No effect was observed in VSMC from ZO. We showed that the lipogenic effects of TO901317observed in VSMC from Zucker control rats are due solely to the nuclear receptor LXRα, PXR agonist had no effect. Conclusion: Lipogenesis is not increased in arterial wall during insulin-resistance and diabetes.