Le diabète de type 2 et le cancer représentent des problèmes majeurs de santé publique. Une cible thérapeutique importante de ces affections est la protéine tyrosine phosphatase PTP1B. Cette dernière est connue pour réguler la voie de l’insuline en déphosphorylant le récepteur de l’insuline, IR ou le substrat du récepteur de l’insuline, IRS. Cependant lesfonctions de PTP1B, le mécanisme par lequel cette phosphatase est régulée restent très ou pas connus. Deux études ont notamment décrites des effets opposés de l’activité de PTP1B suite à la phosphorylation sur résidu de Ser50 de PTP1B par CLK1/CLK2, des kinases LAMMER d’une part et AKT d’autre part. AKT a aussi été montré de phosphoryler la kinase LAMMER CLK1. Par ailleurs, le rôle de PTP1B dans la régulation de la voie Ras/MAPK et donc dans le cancer est un sujet très controversé. L’objectif premier de ce travail de thèse a été d’analyser, le rôle de Ptp61F (l’orthologue de Drosophile de PTP1B) dans la voie de l’insuline de Drosophile, l’interaction entre la phosphatase et la kinase LAMMER de Drosophile, DOA, le rôle de cette phosphatase dans la voie RAS/MAPK. Pour se faire, nous avons utilisé la puissance génétique de laDrosophile pour générer un mutant du gène Ptp61F qui a été caractérisé et son rôle dans les voies de signalisation a été étudié. Cette étude a montrée que, Ptp61F interagit avec IR comme PTP1B chez les mammifères. Elle montre que Ptp61F régule les acteurs clés de la voie de l’insuline Pi3K/Akt. Elle a également montrée que Ptp61F pouvait réguler les fonctions du gène de la kinase LAMMER de Drosphile, Doa. Elle montre enfin que Ptp61F interagit avec nombreuses composantes de la voie RASMAPK de Drosophile (Egfr, Ras, rl (ERK humain)) en réprimant la fonction de chacun de ces gènes et que Rl serait un substrat direct de PTP61F. Les informations selon lesquelles, Ptp61F interagit avec Akt et le gène de la kinaseLAMMER de Drosophile, Doa ont été utilisées dans la deuxième étude pour montrer le rôle que les kinases LAMMER (notamment CLK2, Cdc-like kinase 2) pouvaient jouer dans la voie de signalisation de l’insuline au niveau moléculaire en utilisant les cellules de neuroblastome humain SH-SY5Y. Il en ressort que la kinase CLK2 joue un rôle importantdans cette voie de signalisation. CLK2 est induit par l’insuline et son expression augmente avec le temps. PTP1B interagit in vitro et in vivo avec CLK2. La surexpression de CLK2 induit la baisse de la phosphorylation de AKT par un mécanisme qui pourrait passer par PTP1B, puisque in vitro, CLK2 phosphoryle PTP1B et ce dernier interagit avec AKT in vivo. C’est le résidu de Ser50 de PTP1B qui est phosphorylé et cette phoshphorylation réprime l’activité de PTP1B in vitro. On n’observe cependant pas AKT capable de phosphoryler PTP1B in vitro suggérant que la phosphorylation de PTP1B par AKT serait dépendante du contexte cellulaire. / Type 2 diabetes and cancer represent the major public health problems. One important therapeutic target for these pathologies is the protein tyrosin phosphatase PTP1B. The phosphatase is known to negatively regulates the insulin signaling pathway by dephosphorylating the insulin receptor, IR or the insulin receptor substrate, IRS. However,PTP1B functions and its regulation mechanism remain poorly known. Two studies has notably described opposite effects of PTP1B activity following phosphorylation of its Ser50 residue either by CLK1/CLK2, LAMMER kinases or by AKT. Furthermore, AKT, a main insulin signaling pathway component, has been shown to phosphorylate the LAMMER kinaseCLK1 following insulin stimulation. In addition, the role of PTP1B in the regulation of the RAS/MAPK signaling pathway and hence in cancer is a very controversial subject. The first objective of this work was to analyse, the role of Ptp61F (the Drosophila ortholog of human PTP1B) in the Drosophila insulin pathway, the interaction between the phosphatase and the Drosophila LAMMER kinase gene, Doa, the role of Ptp61F in the RAS/MAPK signaling pathway. To achieve these, we took advantage of the genetic powerful of Drosophila to generate a Ptp61F gene mutant which has been characterized and its role in signaling pathways has been studied. This study showed that Ptp61F interacts with IR like PTP1B in mammals. It shows that Ptp61F regulates key components of insulin signaling pathway Pi3K/Akt. It also shows that Ptp61F is able to regulate the Drosophila LAMMER kinase gene, Doa. Finally, we noted that Ptp61F interacts by inhibiting the activity of severalcomponent of the RAS/MAPK signaling pathway of Drosophila (Egfr, Ras, rl (human ERK)) and conclude that Rl coud be a direct substrate of PTP61F. The data showing that Ptp61F interacts with Akt and the Drosophila LAMMER kinase gene, Doa, were the basis for the second study in order to show the role that the mammal LAMMER kinase CLK2 (Cdc-like kinase 2) could play in the insulin signaling pathway at molecular level using the human neuroblastoma cell line SH-SY5Y. From this second study, we show that CLK2 play an important role in insulin signaling. CLK2 is induced by insulin and its expression increases with time. PTP1B interacts in vivo and in vitro with CLK2. Overexpression of CLK2 impairs AKT phosphorylation by a mechanism which could involved PTP1B, since in vitro, CLK2 phosphorylates PTP1B and the latter interacts withAKT in vivo. It is the Ser50 residue of PTP1B being phosphorylated by CLK2 and this phosphorylation event represses PTP1B activity in vitro. AKT cannot phosphorylates PTP1B in vitro, suggesting that the phosphorylation of PTP1B by AKT could be cellular environment dependant.

A doença cardiovascular (DCV) é a principal causa de mortalidade e morbidade em pacientes portadores de diabetes mellitus tipo 2 (DM 2), estando associada com mais de 80% das mortes nesses pacientes. Portadores de DM 2 têm um risco três vezes maior em relação à indivíduos não diabéticos de desenvolver aterosclerose e suas complicações clínicas como infarto agudo do miocárdio (IAM), acidente vascular cerebral (AVC) e doença vascular periférica. O sistema endócrino da vitamina D regula a diferenciação e a proliferação de vários tipos celulares, além de possuir propriedades antiinflamatórias e antiangiogênicas. Assim, a vitamina D poderia ter um papel protetor contra as doenças degenerativas crônicas como a DCV. Estudos epidemiológicos sugerem que a deficiência de vitamina D está associada com doença arterial coronariana (DAC). As ações da 1,25(OH)2D3 são mediadas pela sua ligação ao seu receptor nuclear (VDR). O objetivo do presente estudo foi investigar as associações de polimorfismos de um único nucleotídeo (SNPs) no gene VDR com DAC em duas coortes de pacientes portadores de DM 2. Um total de 3.137 pacientes provenientes do estudo prospectivo DIABHYCAR (14,8% de incidência de DAC no seguimento) foi estudado. Uma outra coorte (NECKER - COCHIN) independente composta de 713 indivíduos portadores de DM 2 (32,3% dos quais tinham DAC) também foi avaliada. Três SNPs no gene VDR foram genotipados: rs1544410 (BsmI), rs7975232 (ApaI) e rs731236 (TaqI). Uma associação do alelo A de BsmI com casos incidentes de DAC (Hazard Ratio = 1,16; IC 95% = 1,05-1,29; p = 0,002) foi observada, assim como associações do alelo A de BsmI (p = 0,01) e do alelo C de TaqI (p = 0,04) com casos de DAC no início do estudo. O haplótipo AAC (BsmI / ApaI / TaqI) foi significantemente associado com aumento da prevalência de DAC no final do estudo, em comparação com o haplótipo GCT (Odds Ratio = 1,12; IC 95% = 1,02-1,28; p = 0,04). Associações do alelo A de ApaI (p = 0,009) e do alelo C de TaqI (p = 0,05) com DAC foram observadas no estudo transversal da coorte Necker - Cochin. Os resultados obtidos com os haplótipos também foram replicados nessa coorte (Odds Ratio = 1,33; IC 95% = 1,03-1,73; p = 0,03). Em conclusão, o haplótipo composto pelo alelo raro de BsmI, pelo alelo frequente de ApaI e pelo alelo raro de TaqI (AAC) foi associado a um maior risco de DAC em pacientes portadores de DM 2. Este efeito foi independente dos efeitos de outros fatores de risco cardiovasculares / Cardiovascular (CVD) disease is the leading cause of mortality and morbidity in patients with type 2 diabetes being associated with up to 80% of the deaths in these patients. Diabetic patients have a 3-fold higher risk than non diabetic individuals for developing atherosclerosis and its clinical complications such as myocardial infarction, stroke, and peripheral vascular disease. The vitamin D endocrine system regulates the differentiation and replication of several cell types and has antiangiogenic and antiinflammatory properties. Thus, it could have a protective role against chronic degenerative disorders such as CVD. Epidemiological studies suggested that vitamin D deficiency is associated with coronary heart disease. Actions of vitamin D are mediated by the binding of 1,25-(OH)2D3 to a specific cytosolic/nuclear vitamin D receptor (VDR). The present study investigated associations of VDR gene variants with coronary artery disease (CAD) in two cohorts of type 2 diabetic subjects. A total of 3,137 subjects participating in the 6-year prospective DIABHYCAR study (14.8% of CAD incidence at follow-up) were evaluated. An independent, hospital-based cohort (NECKER-COCHIN) of 713 diabetic subjects, 32.3% of whom had CAD, was also studied. Three single nucleotide polymorphisms (SNPs) in the VDR gene were genotyped: rs1544410 (BsmI), rs7975232 (ApaI) and rs731236 (TaqI). In the DIABHYCAR cohort, an association of the A-allele of BsmI with incident cases of CAD was found (Hazard Risk = 1.16; 95% C.I = 1.05-1.29; p = 0.002). Associations were also observed for the A- allele of BsmI (p=0.01) and C-allele of TaqI (p = 0.04) polymorphic variants with baseline cases of CAD. The AAC haplotype (BsmI/ApaI/TaqI) was significantly associated with increased CAD prevalence at the end of the study as compared with the GCT haplotype (Odds Ratio = 1.12; 95% C.I. = 1.02-1.28; p = 0.04). Associations of ApaI (p = 0.009) and TaqI (p = 0.05) alleles with CAD were observed in a cross-sectional study of the NECKER-COCHIN cohort. The haplotype results were also replicated (Odds ratio = 1.33; 95% C.I. = 1.03 - 1.73; p = 0.03). In conclusion, the haplotype composed by the minor allele of BsmI, the major allele of ApaI and the minor allele of TaqI (AAC) was associated with increased risk of CAD in type 2 diabetic patients. This effect was independent from effects of other known cardiovascular risk factors

Patients’ perceptions of health care and support is a key determinant in self-efficacy and active participation in the management of chronic conditions. Nurses play a significant role in Diabetes Self-Management Education by providing clients with tools, empowerment and knowledge to self-manage their condition. The purpose of this study was to examine and describe diabetes patients’ perceptions of nurses’ support for self-management in Botswana. The aim was to improve clients’ skills in self-management and to strengthen diabetes health care management. This study was carried out at the Diabetes Clinic of Excellence, in the city of Francistown, the second largest city in Botswana. The study population comprised of Type 2 Diabetes patients registered at the Diabetes Clinic for their follow-up. All were aged 18 years and above. Three hundred and fifty-four (354) patients participated in the study. The study used a non-experimental, descriptive, quantitative design. Probability sampling method was used to recruit diabetes patients from the selected clinic. Data were were collected using a structured, researcher developed, questionnaire mostly in face-to-face interviews, a few participants completed the questionnaire. The Quantitative data analysis included descriptive and inferential statistics using SPSS (Statistical Package for Social Science Software (version 25). Spearman rho was used to determine statistical correlation between patients’ perceptions and their self-management practices. The findings showed that generally, patients’ perceptions of professional support was positive regarding most of the constructs measured. However, there were areas that showed less satisfaction with the support such as foot-care, risk control, and ability to identify signs of low and high blood sugar level and carrying of Identification Band (ID). Perceptions of nurses’ motivational behaviour showed varied responses. Patients’ self-care activities were sub-optimal and showed some variations which tended to correspond with their perceptions of professionals support. The Spearman's correlation results ranged from strong, moderate, and weak positive correlation. A few demographic variables showed some impact on self-care activities. Based on the findings, the study concludes that professional support through DSME and DSMS, self-management and patients’ perceptions of care play a significant role in diabetes management. / Health Studies / M.A. (Nursing Science)

A obesidade e o Diabetes mellitus tipo 2 se tornaram importantes problemas de saúde pública nos últimos anos. O aumento da prevalência do diabetes está intimamente relacionado ao aumento da prevalência da obesidade. As cirurgias bariátricas surgiram nos últimos cinqüenta anos e vêm se popularizando como uma opção terapêutica efetiva para a redução do peso e controle ou reversão do diabetes no paciente obeso. Dentre as técnicas cirúrgicas disponíveis, o Sleeve gástrico, que era utilizado como parte integrante da técnica de derivação bílio-pancreática ou como primeiro estágio, em pacientes de alto risco, a fim de prepará-los para a cirurgia definitiva, tem sido adotado, por alguns grupos de cirurgiões, como técnica definitiva. Como é recente sua utilização como procedimento bariátrico definitivo, faltam estudos que avaliem sua efetividade. O presente estudo teve por objetivo avaliar o Sleeve gástrico, para o controle ou reversão do Diabetes mellitus tipo 2, no paciente obeso. Para tanto, foi realizada uma revisão sistemática. A busca na literatura resultou em 698 títulos e resumos. Após aplicação dos critérios de inclusão foram recuperados 96 textos completos e incluídos, na revisão sistemática, sete artigos com ensaios clínicos controlados. Foi possível realizar metanálise entre estudos comparando o Sleeve gástrico à derivação gástrica com Y de Roux e com a bandagem gástrica. Foram avaliados os desfechos glicemia de jejum e hemoglobina glicosilada. O resultado da metanálise foi favorável ao Sleeve gástrico, em comparação ao Y de Roux, para o desfecho redução pós-operatória da hemoglobina glicosilada. O Sleeve gástrico se apresenta como mais uma opção terapêutica para a obesidade e correção das co-morbidades associadas. Porém, os resultados são bastante preliminares, e ensaios clínicos controlados, de boa qualidade metodológica, são necessários para melhor avaliação. / Obesity and type 2 diabetes mellitus have become lately important public health problems. The increase of diabetes is closely related to the increase of obesity. The bariatric surgeries appeared in the last fifty years, and have become popular as an effective therapeutic option for reducing weight and for controlling or even reverting diabetes in obese patients. Among the available surgery techniques the sleeve gastrectomy, which was used as an inner part of the bilio-pancreatic diversion technique or as first step in high risk patients in order to prepare them for the final surgery, has been used by some surgeons groups as a definitive technique. As its use as a definitive bariatric procedure is quite recent, there are still not enough studies to evaluate its effectiveness. The present study aims at evaluating the sleeve gastrectomy as a technique for the control or reversion of the type 2 diabetes mellitus in obese patients. Therefore, a systematic review was done. A research in the existing literature presented 698 titles and summaries. After applying the inclusion criteria, 96 complete texts were considered; seven papers with controlled clinic tests were included in the systematic review. This allowed to perform a metanalysis between studies, comparing the sleeve gastrectomy with the Roux-en-Y gastric bypass and with the adjustable gastric band. The comparison was based on the evaluation of the outcome of fast glycemia and glucosided hemoglobin. The result of the metanalysis was favorable to the sleeve gastrectomy, in comparison to the Roux-en-Y gastric bypass, for the closing of the post-surgery reduction of the glycoside hemoglobin. The sleeve gastrectomy thus appears as another therapeutic option towards obesity and correction of the related co-morbidities. But the results are still at an early stage, and so controlled clinical tests with high quality methodology are required in order to achieve a better evaluation.

No description available.

Outre la prescription d’un traitement médicamenteux, la pratique régulière d’une Activité Physique Adaptée (APA)est recommandée aux personnes atteintes d’un diabète de type 2 (DT2), au même titre qu’un nouvel équilibre alimentaire(HAS, 2006). Cette incitation à opérer des changements concrets dans son « style de vie » rencontre des résistances, qui sontaccentuées pour les malades n’ayant pas préalablement construit de dispositions à la pratique physique. Des dispositifsinnovants ont été imaginés dans certaines organisations de santé (réseau de santé, unité transversale d’éducation) pour rendrepossible cet engagement physique et lui permettre de se développer de manière autonome dans le cadre d’« une carrière depratiquant d’APA » au sens où la définit Becker (1985). L’objectif est d’étudier ce processus d’engagement dans ses relationsavec la « trajectoire de maladie » au sens où l’entend Strauss (1985), en rendant compte des différentes étapes de saconstruction. A l’interface de la sociologie de la santé et de la sociologie du sport, l’approche mobilise ainsi des conceptsinteractionnistes. La méthodologie articule une observation de terrain avec 52 récits d’expériences de personnes atteintes demaladie(s) chronique(s) (dont 39 de DT2) qui ont évolué dans deux dispositifs d’APA différents : l’un proposant des séancesthéoriques d’information/explication (n=17), l’autre mettant en place des séances pratiques dans un cycle éducatif en APA puisune orientation vers une association sportive de patients (n=35). L’analyse des récits utilise un logiciel de traitement dedonnées textuelles (Prospéro).Trois types d’engagement ont ainsi été repérés : Le premier s’inscrit dans la « trajectoire demaladie » mais suppose un rapport au corps renouvelé après une remise en question des représentations de l’AP du patient etde ses capacités. Le second ouvre une « carrière de pratiquant d’APA » et ajoute une sociabilité de l’entre-soi, initiée par lesdispositifs « à et via » l’APA. Le troisième consolide cette « carrière » dans une pratique davantage culturelle que médicale,en l’inscrivant dans une sociabilité plus ouverte. Ces trois formes d’engagement se succèdent selon un continuum au coursduquel s’affirment simultanément une recherche croissante d’autonomie par rapport aux prescriptions médicales, une attentiongrandissante au corps et un développement du réseau relationnel. / Alongside prescription medication, it is recommended that type 2 diabetes patients (T2D) carry out regular adaptedphysical activity (APA), in the same way that they should adopt a new dietary balance (HAS 2006). This incitement to bringabout concrete changes in lifestyle comes up against opposition which is compounded where the patient does not have ahistory of regular physical exercise. Some health organizations have devised innovative arrangements such as health networks,or inter-disciplinary patient education units, to make this engagement in physical activity possible, and to enable autonomousdevelopment within the framework of an “APA practitioner career” as defined by Becker (1985). The objective here is tostudy this process of engagement in relation to the “illness trajectory” as understood by Strauss (1985), taking into account thedifferent phases of its course. This approach brings into play the concept of Interactionism at the interface of health sociologyand sport sociology. The methodology used here links fieldwork with 52 personal narratives of people affected by chronicillness (39 of whom have T2D), which have developed through two different APA action plans, the first of which offers theorysessions providing information/explanation (n=17), and the second, the setting up of practical sessions which form a series ofAPA lessons followed by guidance towards a patients’ sports association (n=35). Textual data processing software is used toanalyse the patients’ narratives (Prospéro). Three types of engagement have thus been pinpointed: the first appears in the“illness trajectory” but assumes a new relationship with the body after challenging the patient’s pre-conceived ideas about PAand about his/her own capabilities. The second opens an “APA practitioner career” in the context of socialising with otherpeople like themselves, initiated via the apparatus of APA. The third consolidates this “career” into more of a cultural thanmedical practice, by making it part of a wider group activity. These three types of engagement progress along a continuum,over the course of which the following trends result simultaneously: a growing desire to decrease dependence on medicaladvice, an increase in physical self-awareness, and the development of a network of contacts.

O estresse oxidativo (EO), fenômeno decorrente do desequilíbrio entre a geração de espécies reativas de oxigênio (ERO) e/ou depleção antioxidante (AO), está envolvido na patogênese de diversas doenças e desordens, incluindo o diabetes mellitus tipo 2 (DM2) e a doença periodontal. Uma das conseqüências da geração excessiva das ERO é a destruição tecidual promovida pela peroxidação lipídica, e para combater estes efeitos deletérios, o organismo desenvolveu mecanismos de defesa, como os AO e peroxidases. Objetivos: Avaliar o comportamento dos marcadores de EO na saliva total (SALT) de pacientes com ou sem DM2, portadores ou não de periodontite crônica generalizada (PCG); e verificar e comparar o efeito do tratamento periodontal não-cirúrgico (TPNC) sobre esses marcadores entre os pacientes. Materiais e Métodos: 121 pacientes participaram deste estudo e foram alocados em 4 grupos: diabéticos com PCG (DMPC), diabéticos sem PCG (DM), pacientes sistemicamente saudáveis com PCG (PC) e pacientes sistemicamente saudáveis e sem periodontite (C). Os índices de Placa (IP), nível clínico de inserção (NCI), profundidade clínica de sondagem (PCS) e sangramento à sondagem (SS) foram coletados para a avaliação dos parâmetros clínicos periodontais. Foram também coletadas amostras de sangue periférico e SALT para analisar respectivamente a hemoglobina glicada (HbA1c) e os marcadores de EO, superóxido dismutase (SOD), estado total de antioxidante (TAS) e substâncias reativas do ácido tiobarbitúrico (TBARS) e peroxidase salivar (SPO). As coletas foram realizadas no baseline e após 30 dias do tratamento, previamente à avaliação e reavaliação clínica. Os pacientes DMCP e PC passaram por TPNC, e os DM e C, receberam apenas orientação de higiene oral e profilaxia. Resultados: No baseline, os grupos DMPC e PC apresentaram valores significativamente maiores dos parâmetros clínicos PCS, SS, IP e NCI quando comparados aos grupos DM e C (p<0.05). Após o TPNC, todos esses parâmetros mostraram melhoras significativas (p<0,05). A HbA1c foi significativamente maior no grupo DMPC quando comparado com DM no baseline (p<0,05), no entanto, o TPNC não promoveu melhora significativa após 1 e 3 meses de acompanhamento glicêmico, ainda que, a redução percentual (0,4%) observada possa ser considerada importante. Quanto aos marcadores, no grupo DMPC os valores de TBARS foram significativamente menores comparados aos grupos DM, PC e C (p<0.05), e os de SPO foi significativamente maior em relação aos grupos PC e C (p<0.05), com correlação negativa entre TBARS e SPO (r=-0.35; p=0.002), enquanto que os valores de TAS e SOD não mostraram diferenças significativas entre os grupos estudados (p>0.05). Após o TPNC, os níveis de TBARS aumentaram significativamente, e os de SPO diminuíram significativamente apenas no grupo DMPC (p<0.05 e p<0.05), ao passo que os valores de SOD e TAS permaneceram inalterados nos dois grupos. Conclusão: No baseline, os valores de TBARS e de SPO estiveram alterados nos pacientes DMPC com resultados que mostram correlação inversa entre TBARS/SPO, e após o TPNC houve nova alteração. O comportamento dos marcadores de EO na saliva mostrou a busca pelo ponto de equilíbrio por diversas vias e que este pode ser alterado pelo uso de medicamentos, como a metformina. / Oxidative stress (OS), resulting phenomenon from reactive oxidative species (ROS) and/or antioxidant (AO) depletion imbalance, is involved in many diseases and disorders pathogenesis, including type 2 diabetes mellitus (T2DM) and periodontal disease. One of the ROS excessive generation consequences is the tissue destruction promoted by lipid peroxidation. To strike ROS deleterious effects, the organism developed several defense mechanisms, such as AO and peroxidase systems. Objectives: To evaluate the OS markers behavior in whole saliva (WS) of patients who has or not T2DM, with or without generalized chronic periodontitis (GCP); and to verify and to compare non-surgical periodontal treatment (NSPT) effect on these markers. Material and Methods: 121 patients participated of this study and were allocated in 4 groups: diabetics with chronic periodontitis (DMPC), diabetics periodontally heathy (DM), sistemically healthy with chronic periodontitis (PC) and sistemically and periodontally healthy patients (C). Plaque index (PI), clinical attachment level (CAL), probing depth (PD) and bleeding on probing (BOP) were collected for periodontal clinical paramethers. Peripheral blood and WS samples were collected respectively for glycated hemoglobin (HbA1c) and OS markers, such as superoxide dismutase (SOD), total antioxidant status (TAS), thiobarbituric acid reacting substances (TBARS) and salivary peroxidase (SPO), assessment. Collections were performed at baseline and 30 days after treatment, previously to clinical evaluation and reevaluation. GCP patients (DMPC and PC) were submitted to NSPT, and those with periodontal health (DM and C) received oral hygiene instructions and prophilaxys only. Results: At baseline, DMPC and PC patients showed PD, BOP, PI and CAL higher levels when compared to DM and C groups (p<0.05). After NSPT, all these clinical parameters had significant improvement (p<0.05) in both groups. HbA1c medians were significantly higher in DMPC when compared to DM, at baseline, however, the NSPT did not improved after 1 and 3 months of glicemic control, though percentual reduction (0.4%) observed at clinical point of view might be considered important. As to laboratorial analysis, in DMPC group, TBARS levels were significantly decreased compared to DM, PC and C (p<0.05), and SPO activity were significantly higher than PC and C (p<0.05), with negative correlation between TBARS and SPO (r=-0.35; p=0.002), while TAS levels and SOD activity did not show any significant differences among studied groups (p>0.05). After NSPT, TBARS levels increased significantly, and SPO decreased significantly only in DMPC group (p<0.05 and p<0.05), whereas SOD and TAS values remained unchanged in DMPC and PC groups. Conclusion: At baseline, TBARS levels and SPO activity have been altered in DMPC patients and with results that showed inverse correlation between TBARS/SPO, and after NSPT, another modification occurred. OS markers levels behavior in WS showed a search for balance and this might be changed by medication use, as metformin.

Indivíduos obesos, portadores de diabetes mellitus tipo 2 (DM2), atingem controle glicêmico poucos dias após realização de derivação gástrica em Y de Roux (DGYR), antes mesmo de ocorrer perda de peso significativa. Entre os mecanismos propostos, para explicar o rápido controle glicêmico, a alteração da produção de hormônios gastrintestinais é muito estudada, porém pouco se explorou a expressão tecidual dos genes que os transcrevem. O presente estudo avaliou a resposta, em curto prazo, da expressão gástrica e intestinal de genes que transcrevem peptídeo semelhante ao glucagon 1 (GLP-1), peptídeo insulinotrópico dependente de glicose (GIP), peptídeo tirosina-tirosina (PYY) e grelina, e de níveis sistêmicos desses hormônios à DGYR, bem como, sua correlação com homeostase glicêmica pós-operatória. Mulheres obesas portadoras de DM2 (n=20) foram avaliadas antes e após 3 meses de DGYR. Todas as pacientes foram avaliadas em conjunto e, posteriormente, divididas em dois subgrupos de doentes, determinados pelo grau de remissão pós-operatória de DM2 (parcial ou completa) de acordo com o critério ADA. Biopsias do corpo e fundo gástrico, duodeno, jejuno e íleo foram coletadas por enteroscopia de duplo balão para análise de expressão dos genes GCG (precursor do GLP-1), GIP (precursor do GIP), PYY (precursor de PYY) e GHRL (precursor da grelina). As análises de expressão gênica foram feitas por abordagem global e alvo, por meio das técnicas de microarray e RT-qPCR, respectivamente. Concentrações plasmáticas dos hormônios transcritos pelos genes avaliados também foram dosadas em jejum e após teste de refeição mista, bem como níveis circulantes de peptídeo C, hemoglobina glicada (jejum), glicose, insulina e glucagon (jejum e após teste de refeição mista). Essa abordagem permitiu avaliar a correlação de dados teciduais de expressão gênica e de níveis circulantes de hormônios GI com marcadores sistêmicos de homeostase glicêmica. Após DGYR, alterações significativas na expressão de GCG, GIP, PYY e GHRL ocorreram ao longo dos segmentos GI estudados, em paralelo à melhora significativa de marcadores plasmáticos da homeostase glicêmica. Particularmente, aumento da expressão de GCG e diminuição da expressão de GIP ao longo do intestino foram acompanhados por alterações sistêmicas concordantes dos hormônios por eles transcritos. Essas alterações transcriptômicas se correlacionaram direta (GCG) e inversamente (GIP) com níveis de insulina e glicose, e, inversamente, com níveis de hemoglobina glicada (GCG e GIP). Apenas o hormônio GLP-1 se correlacionou inversamente com níveis de hemoglobina glicada. Além disso, as pacientes com remissão completa de DM2 apresentaram, em relação às medidas pré-operatórias, aumento de GCG no jejuno e íleo, e aumento significativo da área sob a curva de GLP-1 ativo após DGYR. No estômago excluso, ocorreu aumento da expressão de GHRL, mas os níveis sistêmicos de grelina não sofreram alterações significativas no pós-operatório. De acordo com nossos dados, DGYR modificou a expressão de genes que transcrevem hormônios GI capazes de influenciar a homeostase glicêmica. Essas alterações foram acompanhadas por modificações concordantes de níveis sistêmicos das incretinas GLP-1 e GIP e se correlacionaram com marcadores de melhora da homeostase glicêmica. Em conjunto, nossos resultados sugerem que DGYR pode influenciar a homeostase glicêmica por mecanismos que incluem modificação transcriptômica de genes relacionados a hormônios gastrintestinais / Obese subjects with type 2 diabetes mellitus (T2DM) achieve glycemic control few days after Roux-en-Y gastric bypass (RYGB), even before significant weight loss occurs. Among all proposed mechanisms to explain the rapid glycemic control after surgery, alteration of gastrointestinal (GI) hormones production is widely studied, however, GI expression of these genes is still little explored. The present study evaluated the short-term response of gastric and intestinal expression of genes transcribing glucagon-like peptide 1 (GLP-1), glucose-dependent insulinotropic peptide (GIP), tyrosine-tyrosine peptide (PYY), and ghrelin and systemic levels of these hormones after RYGB, as well as their correlation with postoperative glycemic homeostasis. Obese women with T2DM (n = 20) were assessed before and after 3 months of RYGB. All patients were evaluated together and subsequently divided in two subgroups of patients, according the degree of postoperative T2DM remission (partial or complete), following ADA criteria. Biopsies of body and gastric fundus, duodenum, jejunum and ileum were collected by double-balloon enteroscopy for analysis of GCG (precursor of GLP-1), GIP (GIP precursor), PYY (precursor of PYY) and GHRL ghrelin precursor). Gene expression analysis was done by global and target approach, using microarray and RT-qPCR techniques, respectively. Plasmatic concentrations of hormones transcribed by the genes under study were also measured in fasted and after mixed meal test samples, as well as circulating levels of C peptide, glycated hemoglobin (fasting), glucose, insulin and glucagon (fasting and after mixed meal test). This approach allowed the correlation of tissue expression data and circulating levels of GI hormones with systemic markers of glycemic homeostasis. After RYGB, significant changes in GCG, GIP, PYY and GHRL expression were observed along the studied GI segments, in parallel with significant improvement in plasma markers of glycemic homeostasis. In particular, increased GCG expression and decreased GIP expression throughout the small intestine was followed by consistent systemic alterations of its related hormones. These transcriptomic changes are directly (GCG) and inversely (GIP) correlated with insulin and glucose levels and inversely with glycated hemoglobin levels (GCG and GIP). Only GLP-1 was inversely correlated with glycated hemoglobin levels. In addition, regarding the preoperative measurements, patients with complete T2DM remission presented increased GCG in jejunum and ileum, and significant increase in the area under curve of active GLP-1 after RYGB. In the excluded stomach, an increase in GHRL gene expression was recognized, but systemic levels of ghrelin did not changed significantly. According to our data, RYGB altered the expression of genes that transcribe GI hormones related to glycemic homeostasis. These changes were followed by expected modifications of GLP-1 and GIP incretin systemic levels and were correlated with improved glycemic homeostasis markers. Taken together, our results suggest that RYGB can influence glycemic homeostasis by mechanisms that include transcriptomic modification of genes related to gastrointestinal hormones

Alterações morfológicas no tecido ósseo têm sido descritas nos usuários de hipoglicemiantes orais da classe das tiazolidinedionas (TZDs). Hipotetiza-se que alguns genes relacionados com a osteogênese e osteoclastogênese podem ser influenciados pelo tratamento farmacológico, entretanto, o exato mecanismo ainda não está bem esclarecido. O objetivo do estudo foi avaliar o efeito da pioglitazona no remodelamento ósseo através de genes envolvidos na osteoclastogênese em indivíduos recentemente diagnosticados com DM2 e modelos animais, com a finalidade de identificar marcadores genéticos sensíveis de alterações ósseas. Foram convidados para participar do estudo 199 indivíduos (100 diabéticos e 99 normoglicêmicos), no ambulatório de dislipidemias do Instituto Dante Pazzanese de Cardiologia. Os indivíduos diabéticos foram tratados com pioglitazona (15, 30, 45, 45 mg/ dia/ via oral) por 16 semanas. Foram colhidas amostras de sangue, antes e após o tratamento para avaliações laboratoriais, extração de DNA genômico e de RNA total. Os polimorfismos e a expressão do mRNA nas células sanguíneas foram determinados pela PCR em tempo real através do sistema TaqMan®. Para o estudo em modelo animal após a indução da dieta hiperlipídica por 32 semanas, foram utilizados 12 camundongos machos da linhagem C57BL/J6, os quais foram divididos em três grupos: controle (n=4); diabéticos induzidos pela dieta hiperlipídica (DH, n=4) e diabéticos induzidos pela dieta hiperlipídica e tratados com pioglitazona 35mg/Kg/dia por 16 semanas (DHP, n=4). Para os grupos experimentais foram colhidos: amostras de sangue, para exames laboratoriais; fêmures, para a extração do RNA total; e tíbias, para determinação dos parâmetros histomorfométricos. Os pacientes DM2 apresentaram diminuição nas concentrações séricas de osteocalcina e na expressão de OPG e aumento na expressão de VDR em comparação ao grupo NG (p<0,05). A expressão de RANKL e IL6 foi maior entre as mulheres, enquanto que a expressão de PPARG foi maior entre os homens com DM2 em comparação ao grupo NG (p=0,032). Pacientes DM2 antes do tratamento apresentaram glicemia e expressão do mRNA de IL6 negativamente associados ao cálcio ionizado, enquanto que as transcrições de TNFA e VDR foram associadas positivamente e negativamente com bALP respectivamente (p<0,05). O tratamento com pioglitazona reduziu a glicemia de jejum, glicemia pós-prandial, insulina, HOMA-IR, triglicerídeos, VLDL-C, tALP e bALP e aumentou a HDL, tACP, TNF-&#945; e a transcrição de OPG (p<0,05). A glicemia basal associou-se positivamente com o cálcio ionizado. A expressão basal de OPG foi associado negativamente com tALP, enquanto que a expressão basal de TNFA foi associada positivamente com tALP e negativamente com tACP. A expressão basal IL6 foi associada positivamente com tALP, enquanto que a expressão basal de VDR foi associada negativamente com osteocalcina e positivamente com bALP em resposta ao tratamento (p<0,05). O polimorfismo RANK rs1805034 foi associado com redução na transcrição do gene RANK nos indivíduos DM2 e com o remodelamento ósseo após o tratamento com pioglitazona (p<0,05). O polimorfismo RANKL rs9525641 foi associado com aumento da transcrição gênica de RANKL nos indivíduos NG e DM2 e melhora da resposta farmacológica nos indivíduos DM2 tratados com pioglitazona (p<0,05). O polimorfismo rs3102735 do gene OPG foi associado com aumento da formação óssea nos indivíduos DM2 antes e após o tratamento (p<0,05). O genótipo CG do polimorfismo OPG rs2073618 foi associado com alteração da transcrição de OPG no grupo DM2 pré e pós-tratamento (p<0,05). O polimorfismo PPARG rs1801282 foi associado com menor risco para o desenvolvimento de diabetes (p<0,05). O polimorfismo PPARG rs2972162 foi associado com melhora da resistência insulínica nos indivíduos DM2 tratados com pioglitazona (p=0,017). O polimorfismo ESRI rs9340799 foi associado com redução da formação óssea nos indivíduos DM2 (p=0,038). Nos camundongos, após a indução da dieta hiperlipídica por 32 semanas, observou-se aumento do peso, da glicemia, do colesterol total, da expressão do mRNA de RANK, RANKL, IL6 e TNFA em fêmures e aumento de Tb.Sp e diminuição de BV/TV em comparação ao grupo controle (p<0,05). O tratamento com pioglitazona diminuiu a expressão de TNFA (p=0,028). As medidas histomorfométricas não alteraram-se após o tratamento (p>0,05). Os resultados sugerem que o estado hiperglicêmico e o tratamento influenciam os marcadores bioquímicos e moleculares. Os polimorfismos dos genes RANK, RANKL, OPG e ESRI parecem estar envolvidos no remodelamento ósseo independentemente da hiperglicemia e do tratamento e os polimorfismos do gene PPARG parecem estar envolvidos com menor risco para desenvolver diabetes e com a melhora da resistência insulínica em resposta ao tratamento com pioglitazona. / Morphological changes in bone tissue have been reported in users of oral hypoglycemic class of thiazolidinediones (TZDs). It is hypothesized that some genes related to osteogenesis and osteoclastogenesis may be influenced by pharmacological treatment, however, was not aware exact mechanism. The study aims was to evaluate pioglitazone effect on bone remodeling through genes involved in osteoclastogenesis in individuals newly diagnosed with DM2 and animal models, in order to identify sensibles genetics markers of bone alterations. Were invited to participate in study 199 patients (100 diabetics and 99 normoglycemic), in dyslipidemia ambulatory of Institute Dante Pazzanese of Cardiology. Diabetic subjects were treated with pioglitazone (15, 30, 45 or 45 mg /day/oral) for 16 weeks. Blood samples were collected before and after treatment for laboratory evaluations, extraction of genomic DNA and total RNA. Polymorphisms and mRNA expression in blood cells was determined by real time PCR using TaqMan® system. For study in animal model after 32 weeks of fat diet induction, was used 12 male mice C57BL/J6, which were divided into three groups: control (n=4); induced diabetic fat diet (DH, n=4) and induced diabetic fat diet and treated with pioglitazone 35mg/Kg/day for 16 weeks (DHP, n=4). For experimental groups were collected: blood samples for laboratory tests; femurs, for extraction of total RNA; and tibias, to determine histomorphometric parameters. DM2 patients showed decrease in serum osteocalcin and OPG expression and increased VDR expression compared to NG group (p<0.05). RANKL and IL6 expression were higher among women, whereas PPARG expression was higher among men with DM2 compared to NG group (p=0,032). DM2 patients before treatment showed blood glucose and IL6 mRNA expression negatively associated with ionized calcium, whereas TNFA and VDR transcription are positively and negatively associated with bALP respectively (p<0.05). Pioglitazone treatment reduced fasting glucose, postprandial glucose, insulin, HOMA-IR, triglycerides, VLDL-C, tALP and bALP and increased HDL, tACP, TNF-&#945; and OPG transcription (p<0.05). Basal blood glucose was positively associated with ionized calcium. Basal OPG expression was negatively associated with tALP, whereas basal TNFA expression was positively associated with tALP and negatively with tACP. Basal IL6 expression was positively associated with tALP, whereas basal VDR expression was negatively associated with osteocalcin and positively with bALP in response to treatment (p<0.05). RANK rs1805034 polymorphism was associated with RANK gene transcription reduction in subjects with DM2 and bone remodeling after treatment with pioglitazone (p<0.05). RANKL rs9525641 polymorphism was associated with increased RANKL gene transcription in NG and DM2 subjects and pharmacological response improvement in DM2 subjects treated with pioglitazone (p<0.05). OPG rs3102735polymorphism was associated with increased bone formation in DM2 subjects before and after treatment (p<0.05). CG genotype of OPG rs2073618 polymorphism was associated with OPG transcription change in DM2 group before and after treatment (p<0.05). PPARG rs1801282 polymorphism was associated with lower risk for diabetes development (p<0.05). PPARG rs2972162 polymorphism was associated with insulin resistance improvement in DM2 subjects treated with pioglitazone (p=0,017). ESRI rs9340799 polymorphism was associated with reduced bone formation in DM2 subjects (p=0,038). In mice, after 32 weeks of fat diet induction, was observed increase weight, blood glucose, total cholesterol and RANK, RANKL, IL6 and TNFA mRNA expression in femurs and Tb.Sp increase and BV/TV decrease compared to control group (p<0.05). Treatment with pioglitazone decrease TNFA (p=0,028). Histomorphometrics measurements not change after treatment (p>0.05). Results suggest that hyperglycemic state and treatment influence biochemical and molecular markers. RANK, RANKL, OPG and ESRI polymorphisms seens to be involved in bone remodeling regardless of hyperglycemia and treatment and PPARG gene polymorphisms seens to be associated with lower risk for diabetes development and with insulin resistance improvement in response to treatment with pioglitazone.

Le diabète est une maladie chronique avec une progression alarmante. L’insuline-résistance et la diminution de la masse fonctionnelle des cellules beta, associée à l'inflammation des îlots, sont les principaux défauts impliqués dans la pathogenèse du diabète de type 2 (DT2). La compréhension des mécanismes impliqués dans l'inflammation des îlots pancréatiques, et l'identification de cibles moléculaires à visée anti-inflammatoire, sont des approches intéressantes pour le traitement du diabète. La glycogène synthase kinase 3 (GSK3), est une sérine-thréonine kinase qui régule des fonctions cellulaires essentielles. Cette enzyme a été récemment décrite comme un régulateur important de l'inflammation dans différentes conditions pathologiques. Cependant, l'implication potentielle de GSK3beta dans l'inflammation des îlots au cours du diabète reste inexplorée. Le but de ce travail était d'étudier l'implication de GSK3beta dans l'inflammation des îlots pancréatiques et d'évaluer l'impact de l'inhibition de GSK3beta dans l’amélioration de l’hyperglycémie du rat diabétique Goto-Kakizaki. Le rat Goto-Kakizaki (GK) est un modèle spontané de DT2, avec une hyperglycémie chronique apparaissant au sevrage, une masse beta cellulaire réduite et une altération profonde de la sécrétion d'insuline en réponse au glucose. Peu après le sevrage, l'inflammation se développe dans les îlots du rat GK et participe au dysfonctionnement des cellules beta. Nous avons traité les rat GK mâles avec du chlorure de lithium (LiCl), un inhibiteur de GSK3. Le traitement chronique de jeunes rats GK a permis d’éviter l’installation de l’hyperglycémie chronique qui se développe normalement dans ce modèle chez les adultes. A la fin du traitement, la glycémie basale des rats GK traités par le LiCl était fortement réduite, en comparaison avec celle des rats GK non traités. Ces améliorations étaient associées à une réduction de l'expression des cytokines et des chimiokines pro-inflammatoires dans les îlots. L’inhibition de GSK3 a également diminué la fibrose des îlots et rétabli partiellement la sensibilité à l’insuline et la sécrétion d'insuline induite par le glucose chez les rats GK. De plus, des études ex vivo sur des îlots humains et des îlots de rats Wistar, exposés à un environnement inflammatoire en culture, ont révélé l'implication directe de GSK3 dans la réponse inflammatoire autonome des îlots. Ceci était entre autres associée à l’activation du facteur de transcription STAT3. En conclusion, nous montrons pour la première fois que GSK3beta est impliquée dans l’inflammation des îlots pancréatiques humains et de rongeurs. L’inhibition de GSK3beta atténue fortement l’inflammation insulaire, et prévient l’installation de l’hyperglycémie chronique chez le rat GK. L’ensemble des résultats de ce travail nous permet de proposer GSK3beta comme une cible potentielle pour le développement de traitements anti-inflammatoires dans le contexte du diabète de type 2 / Diabetes Mellitus (DM) is a chronic disabling disease with epidemic dimension. It is now established that islet inflammation is associated with defective functional beta cell mass in type 2 diabetes. The understanding of the mechanisms that govern diabetes-associated inflammation in pancreatic islets, and the identification of molecular targets to dampen inflammation are important steps to address this pathological condition. GK rat is a spontaneous model of type 2 diabetes with impaired beta cell function and mass, closely associated with islet inflammation. Glycogen Synthase Kinase 3 (GSK3) is a multi-tasking serine-threonine kinase which regulates crucial cellular functions. In recent years, GSK3beta has been found to be an important regulator of inflammation in different diseased conditions. However, the potential role of GSK3beta in the context of islet inflammation remains unexplored. In this study, we tested the potential of lithium, an inhibitor of GSK3, in improving islet inflammation and glucose metabolism in the GK rat. In vivo, treatment of young GK rats prevented the development of overt diabetes which normally occurs in adult individuals. Lithium improved the glycemic status of the GK rats after few weeks of treatment. At the end of the protocol, GK rats treated with lithium had a blood glucose levels that were significantly lower than that of age-matched untreated GK rats, which were overtly diabetic at this stage. Lithium treatment resulted in reduced expression of pro-inflammatory cytokines and chemokines, decreased fibrosis and reduced macrophage infiltration in the islets. Lithium partially restored the pancreatic insulin content, the insulin sensitivity and the glucose induced insulin secretion in the GK rats. Moreover, ex vivo studies in non-diabetic human and rat islets exposed to inflammatory environment in culture, revealed the direct implication of GSK3 in the islet autonomous inflammatory response. Moreover, we showed that GSK3 controls the islet inflammatory response at least in part by regulating the activity of the pro-inflammatory transcription factor STAT3. Taken together, our results identified GSK3 as a viable target to treat diabetes-associated inflammation, and could have potential clinical application in the treatment of diabetes and metabolic syndrome