The HIF prolyl 4-hydroxylases (HIF-P4H) control hypoxia-inducible factor (HIF), a powerful mechanism regulating cellular adaptation to decreased oxygenation. The gastrointestinal epithelium subsists in “physiological hypoxia” and should therefore have an especially well-designed control over this adaptation. Thus, we assessed the absolute mRNA expression levels of the HIF pathway components, Hif1a, HIF2a, Hif-p4h-1, 2 and 3 and factor inhibiting HIF (Fih1) in murine jejunum, caecum and colon epithelium using droplet digital PCR.We found a higher expression of all these genes towards the distal end of the gastrointestinal tract. We detected mRNA for Hif-p4h-1, 2 and 3 in all parts of the gastrointestinal tract. Hif-p4h-2 had significantly higher expression levels compared to Hif-p4h-1 and 3 in colon and caecum epithelium. To test the roles each HIF-P4H isoform plays in the gut epithelium, we measured the gene expression of classical HIF target genes in Hif-p4h-1/, Hif-p4h-2 hypomorph and Hif-p4h-3/ mice. Only Hif-p4h-2 hypomorphism led to an upregulation of HIF target genes, confirming a predominant role of HIF-P4H-2. However, the abundance of Hif-p4h-1 and 3 expression in the gastrointestinal epithelium implies that these isoforms may have specific functions as well. Thus, the development of selective inhibitors might be useful for diverging therapeutic needs.

Orientador: José Luiz de Mello Nicoletti / Banca: José Luiz de Mello Nicoletti / Banca: Armen Thomassian / Banca: Roberto Pimenta de P. Foz Filho / Resumo: Na prática cotidiana da medicina eqüina, são freqüentes as situações onde é necessária a reposição e ou manutenção do equilíbrio hidroeletrolítico com fluídos e eletrólitos, principalmente em afecções do trato digestório. O presente estudo teve como objetivo avaliar a técnica cirúrgica de fistulização temporária do ceco com a implantação de uma sonda nesta víscera e a utilização desta via para a administração de fluido enteral. Foram utilizados 6 eqüinos adultos entre 350 e 435 kg, machos e fêmeas, que foram submetidos a dois períodos de jejum - hídrico e alimentar - um período sem hidratação e outro período com a hidratação enteral por via intracecal. A solução hidratante foi constituída de 5,7 g de Cloreto de Sódio, 3,78 g de Bicarbonato de Sódio, e 0,37 g de Cloreto de Potássio por litro de água administrado seis vezes ao dia, na base de 50 ml/kg de peso. Ao final do experimento pudemos constatar que a técnica cirúrgica de implantação da sonda é viável e de baixo custo, podendo ser considerada uma opção viável de terapia hídrica, eletrolítica e de nutrição enteral em eqüinos, nas situações em que as vias tradicionais - intravenosa e nasogástrica - se apresentam indisponíveis ou a terapia tradicional se mostra economicamente inviável. / Abstract: In daily equine medical practice is often necessary to re-establish or maintain hemodynamic balance with fluids and electrolytes, mainly due to digestive tract diseases. The objective of this study is to evaluate therapeutic caecal fistulization surgery, with a catheter placement and the main trans and post-operative complications. Six adult horses of both sexes, weighting between 350 and 435kg were used; they were submitted to two periods of total fast - hydric and nutritional - one period without hydration and the other with enteral hydration by intracaecal route. The hydrating solution consisted of 5.7g sodium chloride, 3.78g sodium bicarbonate, and 0.37g potassium chloride per litre of water. We could observe the horse's capacity to tolerate long periods without food and water, and that surgical catheter implant is a viable and low cost option for hydration and electrolyte therapy, and enteral nutrition in horses where traditional routes - intravenous and nasogastric - are not available or traditional therapy is not economically viable. / Mestre

Les spatangoïdes (échinides détritivores fouisseurs) possèdent un volumineux caecum qui s’ouvre au début de l’estomac, le caecum gastrique. Ce caecum est ‘distendu’ :il est toujours gorgé d’un liquide incolore dont la nature est inconnue. Les sédiments ingérés par ces oursins et qui occupent le reste du tube digestif, ne pénètrent jamais dans le caecum. La fonction du caecum gastrique n’est pas claire: il sécréterait des enzymes dans l’estomac, serait un site d’absorption, ou encore abriterait une microflore cellulolytique. En prenant pour modèle l’un des échinides fouisseurs les plus étudiés, Echinocardium cordatum, ce travail tente d’élucider le rôle du caecum gastrique, et s’intéresse plus particulièrement à l’étude de son contenu.<p>Les résultats indiquent que le caecum gastrique d’E. cordatum contient de l’eau de mer. L’entrée d’eau de mer dans le caecum a été visualisée en la colorant et des caractéristiques communes au liquide caecal et à l’eau de mer environnante ont été observées: une même osmolarité, les mêmes particules détritiques en suspension et les mêmes communautés bactériennes. Le caecum gastrique contient de la matière organique en suspension (détritus, bactéries transitoires); il est également absorbant. Ses capacités d’absorption ont été comparées à celles de l’estomac et de l’intestin grâce à un dispositif expérimental particulier :les chambres de Ussing. Les résultats ont montré que les entérocytes du caecum et de l’intestin participent davantage au transfert de glucose vers la cavité coelomique que ceux de l’estomac.<p>Un schéma de la circulation de l’eau de mer dans le tube digestif est proposé. L’eau de mer qui circule à la surface du corps de l’oursin et qui provient de la surface des sédiments atteint la cavité buccale, une circulation entretenue par la ciliature des clavules (piquants ciliés). Le péristaltisme de l’œsophage et celui du siphon assurent l’entrée d’eau de mer dans le tube digestif. Une partie de cette eau entre dans le siphon qui l’amène dans l’intestin d’où elle est entraînée à l’extérieur avec le bol alimentaire. L’eau de mer qui n’est pas prélevée par le siphon peut atteindre l’entrée du caecum gastrique. Un système de gouttières a été mis en évidence à l’entrée du caecum. Il s’étend de l’estomac au début du caecum où les gouttières sont flagellées, et acheminerait l’eau de mer dans la lumière caecale. Les différences de pression osmotique entre le liquide caecal et le liquide cœlomique permettraient le transfert d’eau depuis le caecum vers la cavité cœlomique. Une quantité d’eau similaire devrait alors être éliminée de la cavité coelomique. Cette élimination semble se faire dans le caecum intestinal, l’eau serait ensuite éliminée par l’anus. <p>D’après nos observations, le caecum gastrique pourrait être le site d’une digestion et d’une absorption de la matière organique détritique de l’eau de mer. Si cette hypothèse est exacte, E. cordatum serait alors un détritivore particulièrement ‘complet’, digérant non seulement la fraction détritique des sédiments mais aussi celle en suspension dans l’eau de mer. Ce modèle pourrait correspondre à tous les échinides atélostomes (spatangoïdes & holastéroïdes) qui, outre la présence d’un caecum gastrique bien développé et rempli de liquide, ont en commun d’être fouisseurs, et d’entretenir une circulation d’eau dans leur terrier grâce à des clavules groupés en fascioles.<p><p>Spatangoids (burrowed deposit-feeding echinoids) have a large caecum, which opens at the beginning of the stomach, the gastric caecum. It is always swollen, filled with a colorless liquid whose nature is unknown; sediments ingested by sea urchins fill the rest of the digestive tract but never enter in the caecum. The function of the gastric caecum is unclear: it would secrete enzymes in the stomach, would be a site of absorption, and/or would harbor a cellulolytic microflora. By taking as model one of the most studied burrowing echinoids, Echinocardium cordatum, this study attempts to highlight the role of the gastric caecum by examining its contents.<p>Results indicate that the gastric caecum of E. cordatum contains seawater. Seawater inflow into the caecum was visualized using dye. The caecal liquid and the surrounding seawater were demonstrated to have similar characteristics: the same osmolarity, the same suspended particles and the same bacterial communities. The gastric caecum contains suspended organic matter (detritus, transient bacteria) and is also involved in absorption. Absorption and transfer of glucose were compared between the gastric caecum, the stomach and the intestine, using a particular experimental device: the Ussing chamber. The results showed that the enterocytes of the caecum and of the intestine were more involved in glucose transfer to the coelomic cavity than those of the stomach.<p>Seawater circulation in the digestive tube is tentatively described. Seawater currents along the body of the sea urchin originate from the sediment surface and reach the mouth; this circulation is generated by ciliae of specialized spines, the clavules. Peristalsis of the esophagus and of the siphon induces seawater to enter the mouth and to move along the digestive tube. Part of this water enters the siphon, being then transported to the intestine, and driven outside via the anus. Seawater that has not been taken by the siphon can reach the opening of the gastric caecum. A system of grooves occurring at the entrance of the caecum extends from the anterior stomach to the proximal part of the caecum where it is flagellated; these grooves could transport seawater in the caecal lumen. Differences in osmotic pressures between the caecal liquid and the coelomic liquid could transfer water from the caecum to the coelomic cavity. A similar uptake of water could then be removed from the coelom through the wall of the intestinal caecum, and water be eliminated from the digestive tube via the anus.<p>According to our observations, the gastric caecum could be specialized in digestion and absorption of detrital organic matter occurring in seawater. If this hypothesis is correct, E. cordatum would be a deposit-feeder feeding both on the detritus fraction of the sediments and on that of seawater. This model could fit all Atelostomata echinoids (spatangoids & holasteroids) which, besides the presence of a well-developed gastric caecum filled with liquid, have in common the burrowing behaviour, and the maintenance of seawater currents in their burrows owing to the action of clavules grouped into fascioles.<p><p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished