Das Darmhormon Glucagon-like peptide-1 (GLP-1) ist ein vielversprechendes Therapeutikum in der Behandlung des Typ 2-Diabetes. Allerdings liegen nur wenige Daten über die molekulare Wirkung des Hormons vor. Daher wurden die intrazellulären Effekte, die GLP-1-induzierten Signaltransduktion und Genexpression am Beispiel der pankreatischen ß-Zellinie INS-1 untersucht. In den INS-1-Zellen zeigte sich nach einer GLP-1-Stimulation eine zeit-, dosis- und glukose-abhängige Phosphorylierung von ERK1/2 bzw. der Aktivierung der Transkriptionsfaktoren Elk-1 und CREB. Während GLP-1 die beiden Signalmoleküle bereits nach wenigen Minuten transient aktivierte, führte Glukose zu einer verzögerten, aber anhaltenden Aktivierung. Beide Stimulanzien gemeinsam bewirkten eine synergistische Aktivierung von ERK1/2 und CREB. Sowohl am Mechanismus der glukose- als auch GLP-1-induzierten Aktivierung sind Ca2[plus]-regulierte Signalprozesse in antagonistischer Weise involviert. So führte die Inhibition von CaM-Kinasen und der intrazellulären Ca2[plus]-Erhöhung zu einer Reduktion der GLP-1- und glukosestimulierten ERK1/2-Phosphorylierung, die induzierte Aktivierung von CREB wurde leicht verstärkt. Die Inhibition der PKA und MEK ließen die Rückschlüsse zu, daß die Aktivie-rung der MAPK-Kaskade teilweise durch die PKA vermittelt wird und eine Wechselwirkung zwischen den Kaskaden existiert. Insbesondere die Induktion der frühen Gene junB, c-fos, nur-77 und zif268, die mit der GLP-1-induzierten ERK1/2- und CREB-Phosphorylierung korrelierten, belegten, daß GLP-1 einen Teil seiner Wirkung über diese Signalwege vermittelt. Die Glukosewirkung hinsichtlich der IEG-Induktion war gering. Beide Stimuli gemeinsam führten analog zum synergistischen Effekt auf signaltransduktorischer Ebene zu einer gesteigerten und verlängerten IEG-Expression, die somit die physiologische Bedeutung von GLP-1 als Glukosekompetenzfaktor unterstrichen.

No description available.

Bypass or resection of the stomach and oesophagus, has long been recognised to result in profound changes in the handling of ingested nutrients. This results in significant morbidity after radical surgery for oesophago-gastric cancer, in particular post-prandial hypoglycaemia, altered appetite, early satiety and noxious post-prandial symptoms. By profiling and challenging the gut hormone axis in healthy volunteers and patients who had undergone total or subtotal gastrectomy, or oesophagectomy, this thesis explores the possible causative mechanisms for the challenges faced by this patient population. In the surgical groups, an oral glucose tolerance test (OGTT) resulted in enhanced secretion of satiety and incretin gut hormones (GLP-1, GIP, PYY) and insulin, followed by hypoglycaemia in a cohort of patients. Continuous glucose monitoring of gastrectomy participants over two weeks of normal lifestyle identified an increased incidence of day and night time hypoglycaemia. RNAseq and mass spectrometry based peptidomics of human and murine enteroendocrine cells in the pre- and post-operative populations revealed no significant change in the underlying cellular pathways for nutrient sensing and gut hormone secretion, indicating that the altered hormone secretion is primarily driven by accelerated nutrient transit, rather than adaptive changes in the gut. Finally, specific blockade of the GLP-1 receptor in post-gastrectomy patients using Exendin 9-39 normalised insulin secretion and prevented reactive hypoglycaemia after an OGTT. In conclusion, profound changes in gut hormone secretion as a result of enhanced nutrient transit after foregut surgery likely underlie the early and late post-prandial symptoms seen in this group, and therapies specifically targeting the gut hormone axis, and GLP-1 in particular, could be the first targeted treatments for post-gastrectomy syndromes.

Dans la perspective de trouver de nouvelles thérapies dans le traitement du diabète de type II, non insulino-dépendant, le Glucagon-Like Peptide-1 (GLP-1) constitue un excellent candidat. Si son mécanisme d'action est bien connu, il reste toutefois à résoudre de grands problèmes fondamentaux, avant de le substituer aux molécules utilisées pour une telle pathologie. En particulier, la compréhension de la liaison du GLP-1 à son récepteur demeure un point crucial. Une meilleure connaissance des structures du ligand et du récepteur sont nécessaires. De plus, ce peptide ne peut être utilisé dans sa forme native, dû à une inactivation rapide par les protéases.<br /><br />Pour essayer d'augmenter la stabilité du peptide, et en tenant compte des positions clés définies dans la littérature, plusieurs analogues du GLP-1-(7-37) sont conçus, et synthétisés. Ils possèdent principalement des pharmacomodulations au niveau de la partie N-terminale. Des substitutions sont également réalisées dans la partie centrale du peptide, permettant de vérifier certaines hypothèses concernant sa conformation. Considérant les résultats de liaison et d'efficacité in vitro, certains analogues sont sélectionnés pour des études in vivo d'activité et de stabilité métabolique. Le [a8,desR36]GLP-1-(7-37) se distingue des autres tant par sa grande stabilité que son efficacité, supérieure à la molécule native. Ce composé est en phase de développement pré-clinique.<br /><br />Parallèlement, la conformation de chaque analogue est étudiée (CD, IR) et ainsi, confrontée aux résultats in vitro, il est possible de proposer une conformation bioactive.<br /><br />Enfin, pour appréhender plus en avant les mécanismes de liaison du peptide avec son récepteur spécifique, la modélisation moléculaire du récepteur fait ressortir quelques hypothèses quant à la localisation probable de l'interaction hormone-récepteur. Des analyses biophysiques et la synthèse de fragments du récepteur, ont permis d'étayer de telles hypothèses.

Made available in DSpace on 2015-07-22T20:49:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-06-29 / TEDE / BV UNIFESP: Teses e dissertações

Type 2 diabetes is still rapidly on the rise today, affecting 10.5% of individuals in the United States between the ages 45 to 64 and 18.4% of those between the ages of 65 to 74. In the past two decades, type 2 diabetes has doubled in all age groups. Many adults with type 2 diabetes experience difficulty managing their blood sugars, which can result in a range of further complications. One of the newest treatment options on the market today is a glucagon-like peptide-1 (GLP-1) receptor agonist, Bydureon. Similar to Byetta, Bydureon has a main ingredient of exenatide. It offers once a week dosing as opposed to twice-a-day, which may be more appealing to patients. The purpose of this study was to examine the efficacy of a newly FDA released medication, Bydureon, once weekly dosage in adults with type 2 diabetes. A descriptive, comparative, retrospective study of 35 patients evaluated efficacy by examining Hgb A1C and body mass index in adults with type 2 diabetes at baseline and 3 months after Bydureon was prescribed. Data were collected by a chart review of records in a primary care practice. Results demonstrated a statistically significant difference between baseline to 3 month means in both Hgb A1C (t (34)= -3.05, p=.0044) and BMI (t (34) = -2.86, p = .0072) for patients using Bydureon. Health care providers need to individualize the patients’ plans of care to address multifactorial areas of their diabetes care and provide them with an opportunity to successfully meet their goals. Practitioners must be knowledgeable about the treatment options available, including the newer GLP-1 receptor agonist, Bydureon and its efficacy for adults with type 2 diabetes.

Indiana University-Purdue University Indianapolis (IUPUI) / Recent advancements in the management of systemic glucose regulation in obesity/T2DM include drug therapies designed to utilize components of the incretin system specifically related to glucagon-like peptide 1 (GLP-1). More recently, GLP-1 has been investigated for potential cardioprotective effects. Several investigations have revealed that acute/sub-acute intravenous administration of GLP-1 significantly reduces myocardial infarct size following ischemia/reperfusion injury and improves cardiac contractile function in the settings of coronary artery disease, myocardial ischemia/reperfusion injury, and heart failure. Despite an abundance of data indicating that intravenous infusion of GLP-1 is cardioprotective, information has been lacking on the cardiac effects of iv GLP-1 in the MetS or T2DM population. Some important questions this study aimed to address are 1) what are the direct, dose-dependent cardiac effects of GLP-1 in-vivo 2) are the cardiac effects influenced by cardiac demand (MVO2) and/or ischemia, 3) does GLP-1 effect myocardial blood flow, glucose uptake or total oxidative metabolism in human subjects, and 4) are the cardiac effects of GLP-1 treatment impaired in the settings of obesity/MetS and T2DM. Initial studies conducted in canines demonstrated that GLP-1 had no direct effect on coronary blood flow in-vivo or vasomotor tone in-vitro, but preferentially increased myocardial glucose uptake in ischemic myocardium independent of effects on cardiac contractile function or coronary blood flow. Parallel translational studies conducted in the humans and Ossabaw swine demonstrate that iv GLP-1 significantly increases myocardial glucose uptake at rest and in response to increases in cardiac demand (MVO2) in lean subjects, but not in the settings of obesity/MetS and T2DM. Further investigation in isolated cardiac tissue from lean and obese/MetS swine indicate that this impairment in GLP-1 responsiveness is related to attenuated activation of p38-MAPK, independent of alterations in GLP-1 receptor expression or PKA-dependent signaling. Our results indicate that the affects of GLP-1 to reduce cardiac damage and increase left ventricular performance may be impaired by obesity/MetS and T2DM.

[pt] Esta dissertação visa a aplicar uma metodologia para avaliar as diferenças máximas permissíveis nas operações de transferência de custódia de GLP para cada operação em particular, utilizando testes de hipóteses baseados na incerteza de medição, em contraponto à utilização de valores pré-estabelecidos em normas e regulamentos técnicos metrológicos. Tais documentos propõem um valor único de incerteza máximas nas medições de transferência de custódia, sem estabelecer diferenças máximas permissíveis nas operações entre dois sistemas, cabendo aos fornecedores e clientes negociar este parâmetro, o que frequentemente causa disputas. Os resultados mostraram que, em um sistema isolado, ao contrário de outros métodos, as condições ambientais não influenciam o método de medição de massa e sua incerteza. Nenhuma diferença máxima permissível de massa transferida de GLP entre caminhões-tanque e estações de medição excede 1,0 porcento; em contraste, quando uma esfera de armazenamento é parte de um dos sistemas de medição, este parâmetro depende completamente das condições operacionais e definitivamente um valor único não deve ser considerado. / [en] This dissertation aims to apply a methodology to assess the maximum allowable differences in LPG custody transfer operations for each particular operation, using hypothesis tests based on measurement uncertainty, as opposed to using pre-established values in metrological standards and technical regulations. Such documents propose a single value for maximum uncertainty in custody transfer measurements, without establishing maximum permissible differences in operations between two systems, leaving it up to suppliers and customers to negotiate this parameter, which often causes disputes. The results showed that in an isolated system, unlike other methods, environmental conditions do not influence the mass measurement method and its uncertainty. No maximum permissible difference of transferred mass of LPG between tank trucks and measuring stations exceeds 1.0 percent; in contrast, when a storage sphere is part of one of the measuring systems, this parameter completely depends on the operating conditions and definitely a single value should not be considered.

Tesis por compendio / [ES] La obesidad es un gran reto de salud pública que ha alcanzado proporciones epidémicas. El entorno "occidentalizado" en el que vivimos, caracterizado por la accesibilidad a alimentos hipercalóricos, contribuye al desequilibrio crónico entre energía ingerida y gasto energético que causan la obesidad. Las intervenciones conductuales diseñadas para la pérdida de peso tienen limitada efectividad a largo plazo, por lo que existe una urgente necesidad de desarrollar estrategias más eficaces y seguras para prevenir y tratar la obesidad y sus comorbilidades. El desarrollo de estrategias terapéuticas dirigidas al intestino para mejorar la salud metabólica requiere un conocimiento en profundidad de las vías de señalización neuroendocrina intestinal que regulan el la ingesta y el equilibrio energético. El objetivo de esta tesis ha sido profundizar en las interacciones intestino-cerebro implicadas en el control de la homeostasis energética, incluyendo los componentes endocrinos, neurales y la microbiota intestinal, en el contexto del desarrollo de la obesidad inducida por una dieta hipercalórica. En los Capítulos 1 y 2 hemos explorado nuevas funciones de las neuronas sensoriales aferentes que expresan el canal de sodio Nav1.8 en el control de la homeostasis energética, considerando las diferencias entre sexos. Hemos generado un modelo de ratón carente de las neuronas Nav1.8+ mediante ablación con toxina diftérica. En el Capítulo 1 hemos demostrado que las neuronas Nav1.8+ son indispensables para regular específicamente según el sexo las vías neurales y endocrinas implicadas en la homeostasis energética. En hembras, la ablación de estas neuronas mejora la regulación de la glucosa postprandial potenciando la señalización enteroendocrina de GLP-1 y acelera el tránsito intestinal, mientras que en machos induce resistencia al aumento de peso inducido por una dieta obesogénica. En el Capítulo 2 hemos demostrado que, en machos, la ablación de las neuronas Nav1.8+ altera el control coordinado de la ingesta i las variaciones de peso diarias, además de alterar la señalización enteroendocrina y las oscilaciones diarias de la microbiota intestinal en respuesta al estado nutricional (ayuno/ingesta), y perturbar la homeostasis del sistema inmune intestinal. En el capítulo 3, hemos usado un modelo de ratón con obesidad inducida por dieta para explorar los mecanismos por los cuales Phascolarctobacterium faecium DSM 32890, una cepa bacteriana intestinal aislada de humanos metabólicamente sanos, previene la obesidad modulando la ingesta. La administración de P. faecium reduce la ingesta calórica gracias a la hipersecreción de la hormona gastrointestinal saciante el PYY. Independientemente de sus efectos anorexigénicos, la bacteria ejerce sus beneficios metabólicos estimulando el tránsito intestinal y reduciendo la absorción intestinal de lípidos, evitando la acumulación de grasa corporal. En conclusión, esta tesis doctoral proporciona evidencia preclínica que contribuye a una comprensión más precisa de las vías neuroendocrinas que comunican el intestino y el cerebro, y del papel que tiene la microbiota intestinal en la regulación de la ingesta y el gasto energético. Destacamos la importancia de las neuronas sensoriales aferentes Nav1.8+ en la detección de estímulos intestinales por quimiorreceptores para regular el balance energético en ambos sexos, lo cual abre una nueva línea de investigación para diseñar herramientas de neuromodulación de las neuronas Nav1.8+ con el fin de prevenir y tratar los trastornos metabólicos inducidos por la dieta, de forma específica para cada sexo. También destacamos que P. faecium es una bacteria candidata como probiótico de nueva generación, ya que modula el sistema enteroendocrino del hospedador y previene la obesidad en un modelo preclínico. En conjunto, estos hallazgos proporcionan una base para el desarrollo de estrategias terapéuticas basadas en el intestino dirigidas a combatir la obesidad y comorbilidades asociadas. / [CA] L'obesitat és un gran repte de salut pública que ha assolit proporcions epidèmiques. L'entorn "occidentalitzat" en el que vivim, caracteritzat per l'accessibilitat a aliments hipercalòrics, contribueix al desequilibri crònic entre energia ingerida i despesa energètica que causen l'obesitat. Les intervencions conductuals dissenyades per a la pèrdua de pes tenen una eficàcia limitada a llarg termini, per la qual cosa hi ha una necessitat urgent de desenvolupar estratègies més eficaces i segures per a prevenir i tractar l'obesitat i les seues comorbiditats. El desenvolupament d'estratègies terapèutiques dirigides a l'intestí per a millorar la salut metabòlica requereix un coneixement en profunditat de les vies de senyalització neuroendocrina intestinal que regulen la ingesta i l'equilibri energètic. L'objectiu d'aquesta tesi ha sigut aprofundir en les interaccions intestí-cervell implicades en el control de l'homeòstasi energètica, incloent els components endocrins, neurals i la microbiota intestinal, en el context del desenvolupament de l'obesitat induïda per una dieta hipercalòrica. En els Capítols 1 i 2 hem explorat noves funcions de les neurones sensorials aferents que expressen el canal de sodi Nav1.8 en el control de l'homeòstasi energètica, considerant les diferències entre sexes. Hem generat un model de ratolí mancat de les neurones Nav1.8+ mitjançant ablació amb toxina diftèrica. En el Capítol 1 hem demostrat que les neurones Nav1.8+ són indispensables per a regular, específicament segons el sexe, les vies neurals i endocrines implicades en l'homeòstasi energètica. En femelles, l'ablació d'aquestes neurones millora la regulació de la glucosa postprandial potenciant la senyalització enteroendocrina de GLP-1 i accelera el trànsit intestinal, mentre que en mascles indueix resistència a l'augment de pes induït per una dieta obesogènica. En el Capítol 2 hem demostrat que, en mascles, l'ablació de les neurones Nav1.8+ altera el control coordinat de la ingesta i les variacions de pes diàries, a més d'alterar la senyalització enteroendocrina i les oscil·lacions diàries de la microbiota intestinal en resposta a l'estat nutricional (dejuni/ingesta), i pertorbar l'homeòstasi del sistema immunitari intestinal. En el capítol 3, hem utilitzat un model de ratolí amb obesitat induïda per dieta per explorar els mecanismes pels quals Phascolarctobacterium faecium DSM 32890, una soca bacteriana intestinal aïllada d'humans metabòlicament sans, prevé l'obesitat modulant la ingesta. L'administració de P. faecium redueix la ingesta calòrica gràcies a la hipersecreció de l'hormona gastrointestinal saciant PYY. Independentment dels seus efectes anorexigènics, el bacteri exerceix els seus beneficis metabòlics estimulant el trànsit intestinal i reduint l'absorció intestinal de lípids, evitant l'acumulació de greix corporal. En conclusió, aquesta tesi doctoral proporciona evidència preclínica que contribueix a una comprensió més precisa de les vies neuroendocrines que comuniquen l'intestí i el cervell, i del paper que té la microbiota intestinal en la regulació de la ingesta i la despesa energètica. Destaquem la importància de les neurones sensorials aferents Nav1.8+ en la detecció d'estímuls intestinals per quimioreceptors per a regular l'equilibri energètic en ambdós sexes, que obri una nova línia d'investigació per a dissenyar ferramentes de neuromodulació de les neurones Nav1.8+ amb la finalitat de prevenir i tractar els trastorns metabòlics induïts per la dieta, de forma específica per a cada sexe. També destaquem que P. faecium és un bacteri candidat com a probiòtic de nova generació, ja que modula el sistema enteroendocrí de l'hoste i prevé l'obesitat en un model preclínic. En conjunt, aquests troballes proporcionen una base per al desenvolupament d'estratègies terapèutiques basades en l'intestí dirigides a combatre l'obesitat i comorbiditats associades. / [EN] Obesity is a major global public health challenge that has reached epidemic proportions. Besides its profound impact on health and well-being, this metabolic disorder represents a significant economic burden to society. Our westernized environment where high-calorie foods are readily available, represents a major driver of the chronic imbalance between energy intake and energy expenditure that cause obesity. The limited effectiveness of behavioral interventions to manage long-term weight loss highlights the urgent need to develop more effective and minimally invasive approaches to prevent and treat obesity and its comorbidities. The development of gut-targeted therapeutic strategies to improve metabolic health requires a comprehensive understanding of the gut neuroendocrine signaling pathways that, in interaction with the gut microbiota, control feeding behavior to ultimately maintain energy balance. The aim of this thesis has been to gain insight into gut-brain interactions, including those mediated by endocrine, neural and gut microbial components, involved in the control of energy homeostasis, with a focus on obesogenic diet-related dysfunctions that increase susceptibility to develop obesity. In Chapters 1 and 2, we have investigated novel functions of sensory afferent neurons expressing the sodium channel Nav1.8 in the control of energy homeostasis, considering sex-specificities, by generating a mouse model lacking Nav1.8+ neurons through a diphtheria toxin ablation strategy. In Chapter 1, we show that Nav1.8+ neurons are required to control neural and endocrine pathways involved in energy homeostasis in a sex-specific manner. Specifically, ablation of Nav1.8+ neurons in females improves postprandial glucose regulation by enhancing glucagon-like peptide-1 enteroendocrine signaling and accelerating intestinal transit, whereas in males it induces resistance to weight gain in response to an obesogenic diet. To further explore the role of Nav1.8+ neurons in controlling food intake and pre- and post-prandial daily rhythms that influence metabolic phenotype, in Chapter 2 we show in males that ablation of Nav1.8+ sensory neurons impairs the coordinated control of food intake and body weight fluctuations throughout the day. The loss of these neurons also alters the physiological enteroendocrine signaling and daily gut microbiota oscillations in response to the nutritional status (fasting/refeeding cycles) and disrupts intestinal immune homeostasis. In Chapter 3, we used a diet-induced obese mouse model to investigate the mechanisms by which Phascolarctobacterium faecium DSM 32890, a gut bacterial strain isolated from metabolically healthy humans, prevents obesity by modulating food intake. We show that administration of P. faecium reduces caloric intake by promoting hypersecretion of a satiating gastrointestinal hormone, the peptide YY (PYY). Independently of its anorexigenic effects, the bacterium exerts its metabolic benefits via complementary mechanisms, specifically by stimulating intestinal transit and reducing intestinal lipid absorption, thereby preventing body fat accumulation. In conclusion, this doctoral thesis provides preclinical evidence for a better understanding of gut-to-brain neuroendocrine pathways and the role of gut microbiota in the regulation of food intake and energy expenditure. We highlight the importance of Nav1.8+ sensory afferent neurons in gut chemosensing for maintaining energy balance in both sexes, which prompts novel research lines and opportunities to design of sex-specific neuromodulation tools targeting Nav1.8+ neurons for prevention and treatment of diet-induced metabolic disorders. We also highlight that P. faecium is a promising next-generation probiotic candidate, as it modulates the host enteroendocrine system and prevents obesity in a preclinical model. Overall, our findings contribute to the development of gut-based therapeutic strategies to combat obesity and associated comorbidities. / This study has been funded by the European Union 7th Framework Program through the MyNewGut project (Grant agreement No. 613979) and Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No. 797297 (MRP), the Spanish Ministry of Science and Innovation (Grant PID2020-119536RB-I00), the European Commission – NextGenerationEU, through the CSIC Interdisciplinary Thematic Platform (PTI+) NEURO- AGING+ (PTI-NEURO-AGING+)”. The grant of the Spanish Ministry of Science and Innovation (MCIN/AEI) to IATA-CSIC as Accredited Center of Excellence (CEX2021-001189-S/ MCIN/AEI / 10.13039/501100011033) is acknowledged. / Bullich Vilarrubias, C. (2024). Influence of gut-to-brain neuroendocrine pathways and intestinal microbiota on energy homeostasis [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/207342 / Compendio