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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
41

Production et hydrolyse des amides : mécanismes chimiques, isotopie et applications : étude de la glutamine synthétase / Production and hydrolysis of amide : chemical mechanisms, isotopy and applications : study of glutamine synthetase

Mauve, Caroline 15 December 2014 (has links)
La nutrition azotée des bactéries et des plantes est actuellement un sujet de grande importance, notamment pour comprendre comment améliorer les voies métaboliques aboutissant à l’assimilation de l’azote et à plus grande échelle, optimiser des apports d’engrais et augmenter le rendement des cultures. Dans ce contexte, la réaction d’amidation catalysée par la glutamine synthétase (GS), qui fixe l’ammonium (NH₄)⁺ en glutamine, est cruciale car elle est à la fois le point d’entrée de l’azote dans les végétaux, et une étape-clef du recyclage de l’azote (en particulier, NH₄⁺ photorespiratoire). Dans cette étude, nous nous sommes intéressés à la cinétique enzymatique et au mécanisme chimique de la GS. Des systèmes analytiques (HPLC, RMN , GC-MS) ont  été optimisés pour permettre la mesure de l’activité enzymatique in vitro et pour réaliser des analyses par spectrométrie de masse à ratio isotopique. Avec ces techniques, nous avons pu regarder précisément les effets isotopiques ¹²C/¹³C, ¹⁴N/¹⁵N et H₂O/D₂O (solvant) lors de la catalyse, en utilisant la GS d’E. coli et d’Arabidopsis thaliana (GS1,2). Nos résultats montrent qu’il n’y a pas d’effet isotopique ¹²C/¹³C, mais qu’il y a un fractionnement ¹⁴N/¹⁵N de »16‰. En outre, il y a un effet inverse du solvant (réaction 1.5 à 2 fois plus rapide dans D₂O).  Cela suggère que la création de la liaison C----N (amidation) est partiellement limitante (engagement catalytique de »14% seulement) et que le réseau de ponts hydrogènes dans le site actif est crucial pour déterminer la vitesse de la réaction. L’apparition d’effets ¹⁴N/¹⁵N inverses dans certaines circonstances et les effets drastiques causés par une substitution du cofacteur métallique (Mg²⁺) suggèrent en outre que l’étape d’amidation peut être réversible et que la coordination par un métal joue un rôle très important pour stabiliser les intermédiaires de la réaction, en interaction avec le solvant. Ainsi, dans son solvant naturel qu’est H₂O, la GS réalise une réaction ‘chimiquement difficile’ (barrière énergétique élevée de l’amidation) rendue possible par le clivage de l’ATP et son caractère exergonique. / Nitrogen nutrition in bacteria and plants is currently an important topic, in particular to identify key points for metabolic improvements in N assimilation and more generally, to optimize fertilization and crop yield. In such a context, the amidation reaction catalyzed by glutamine synthetase (GS), which fixes ammonium (NH₄)⁺ into glutamine, is of crucial importance since it both represents the N entry in plants and the main step of N recycling (such as photorespiratory (NH₄)⁺. Here, we examined GS kinetics and chemical mechanism. Analytical methods (HPLC, NMR, GC-MS) have been set up so as to measure in vitro activities and isotopic abundance by isotope ratio mass spectrometry. These gave access to isotope effects (¹²C/¹³C, ¹⁴N/¹⁵N et H₂O/D₂O – solvent) during catalysis, with the GS from either E. coli or A. thaliana (GS1,2). Our results show that there no ¹²C/¹³C isotope effect but there is significant ¹⁴N/¹⁵N isotope fractionation of ca. 16‰. In addition, there is an inverse solvent isotope effect (reaction 1.5 to 2 times faster in D₂O). This suggests that forming the C----N bond (amidation) is partially rate-limiting (catalytic commitment of ca. 14% only) and the H-bond network in the active site is of substantial importance for the reaction rate. The occurrence of inverse ¹⁴N/¹⁵N isotope effects under certain circumstances as well as the drastic impact of changing the metal cofactor (Mg²⁺)) indicate that the amidation step can be reversible and that the coordination by the metal plays a key role in stabilizing reaction intermediates, by interfacing the solvent. In other words, in its natural solvent H₂O, the GS catalyses an intrinsically ‘difficult’ reaction (high energy barrier of amidation) made possible by both ATP cleavage and its exergonic nature.
42

Neuropathogenic mechanisms of feline immunodeficiency virus infection

Buck, Wayne R. 04 March 2004 (has links)
No description available.
43

Partial purification and characterization of selected enzymes of bovine nitrogen metabolism : comparison of the Nguni and Hereford breeds

Mathomu, Lutendo Michael 11 1900 (has links)
Ruminant animals consuming low N-diet have been reported to have increased urea reabsorption with the Nguni being categorized as N-recycling ruminant. The enzymes associated with N-cycling are hypothesized to contribute to survival of the Nguni in harsh conditions. Enzymes responsible for such a function needed to be characterized in order to determine their effect in the functioning of the Nguni as opposed to Hereford breed. Crude enzymes from both breeds were separated from most or some contaminants by sephadex G-25, DEAE sephacel, and different affinity column chromatography. CPS and GDH were successfully purified and characterized by LC-MS/MS and further analysed by ProteinPilot™, blasted and matched >95% with those of Bos Taurus. Comparison of characterized enzymes and those which failed to ionise such as ARG, GS and GA was done using kinetics and graphs annotating specific activities. Partial purification and characterization was in part achieved. / Life & Consumer Sciences / M. Sc. (Life Sciences)
44

Repression der cytosolischen GS1 von Zuckerrüben (Beta vulgaris L. var. altissima) durch Antisense-DNA-Konstrukte / Repression of the cytosolic GS1 from sugar beet (Beta vulgaris L. cv. altissima) by using antisense DNA constructs

Hoffmann, Guido Wolf 21 June 2000 (has links)
No description available.
45

Les neutrophiles ne sont pas résistants aux glucocorticoides

Hirsch, Gaëlle 07 1900 (has links)
Les neutrophiles sont généralement considérés résistants aux glucocorticoïdes. Cependant, peu d’études comparant l’effet de ces drogues sur les neutrophiles et les autres leucocytes sanguins (monocytes, lymphocytes et éosinophiles) ont été rapportées. Dans notre étude, nous avons évalué la réponse aux glucocorticoïdes de ces deux populations cellulaires chez le cheval et l’homme. Les cellules, préalablement isolées du sang de 6 chevaux et 4 sujets humains sains, ont été incubées pendant 5 h en présence de lipopolysaccharide (LPS; 100 ng/mL) seul ou combiné avec de l’hydrocortisone, de la prednisolone ou de la dexaméthasone (10-8M et 10-6M). L’expression d’ARNm pour l’IL-1β, le TNF-α, l’IL-8, la glutamine synthétase et le récepteur α des glucocorticoïdes (GR-α) a été quantifiée par qPCR. Les neutrophiles équins ont également été incubés pendant 20 h en présence de ces 3 glucocorticoïdes et la survie cellulaire a été évaluée par cytométrie de flux et microscopie optique. Nous avons démontré que les glucocorticoïdes inhibaient l’expression des gènes pro-inflammatoires induite par le LPS pour les deux populations cellulaires chez les deux espèces étudiées. L’expression de la glutamine synthétase était également significativement augmentée par les glucocorticoïdes chez les neutrophiles et les autres leucocytes sanguins équins. De manière générale, l’intensité de la réponse aux glucocorticoïdes s’est avérée similaire dans les 2 populations leucocytaires et chez les deux espèces. Les glucocorticoïdes augmentaient également la survie des neutrophiles équins, phénomène également rapporté dans d’autres espèces. Ainsi, les glucococorticoïdes exercent des effets d’intensité comparable sur les neutrophiles et les autres leucocytes sanguins. Nous spéculons que la faible réponse à la corticothérapie observée lors de maladies inflammatoires chroniques neutrophiliques comme l’asthme sévère ou la Maladie Pulmonaire Obstructive Chronique (MPOC) ne s’explique pas par une corticorésistance intrinsèque des neutrophiles. / Neutrophils are generally considered resistant to glucocorticoids compared to other inflammatory cells. However, there are few studies comparing the effects of glucocorticoids in neutrophils and those of other blood leukocytes (monocytes, lymphocytes and eosinophils). In our study, we assessed glucocorticoid-responsiveness in equine and human peripheral blood neutrophils and in neutrophil-depleted leukocytes. Cells were isolated from 6 healthy horses and 4 human healthy subjects. They were incubated for 5 h with or without lipopolysaccharide (LPS; 100 ng/mL) alone or combined with hydrocortisone, prednisolone or dexamethasone (10-8M and 10-6M). IL-1β, TNF-α, IL-8, glutamine synthetase and Glucocorticoid Receptor α (GR-α) mRNA expression was quantified by qPCR. Equine neutrophils were also incubated for 20 h with or without the three glucocorticoids and cell survival was assessed by flow cytometry and light microscopy. We found that glucocorticoids down-regulated LPS-induced pro-inflammatory mRNA expression in both cell populations and species. These drugs also significantly increased glutamine synthetase gene expression in both equine cell populations. The magnitude of glucocorticoid response was generally similar in both cell populations and species. As reported in other species, glucocorticoids significantly increase the survival in equine neutrophils. Based on these results, it appears that glucocorticoids exert effects of similar magnitude on neutrophils and on other blood leukocytes. We speculate that the poor response to glucocorticoids observed in some chronic neutrophilic human diseases such as severe asthma or Chronic Obstructive Pulmonary Disease (COPD) is not explained by an inherent attenuated response of neutrophils to these drugs.
46

Aspectos metabólicos, nutricionais e produtivos de cultivares de Brachiaria e Panicum visando eficiência no uso do nitrogênio / Metabolic, nutritional and production aspects of Brachiaria and Panicum cultivars aiming nitrogen use efficiency

Garcez, Tiago Barreto 01 October 2013 (has links)
O melhor entendimento sobre os aspectos metabólicos, nutricionais e produtivos das gramíneas forrageiras quanto ao uso do nitrogênio é necessário para aumentar a eficiência de uso do nutriente e para a sustentabilidade da produção pecuária. Com isso, objetivou-se analisar alguns cultivares de gramíneas forrageiras utilizados em pastagens no Brasil, quanto a: I) modificações morfológicas e produtivas da parte aérea e das raízes; II) formas de partição do nitrogênio absorvido para melhorar a eficiência de uso do nitrogênio e III) forma de assimilação do nitrogênio absorvido, sendo avaliados pelas variáveis: produção de massa seca, números de folhas e perfilhos, área foliar, concentração e conteúdos de nitrogênio total, nitrato e amônio, eficiência de uso do nitrogênio, atividades das enzimas nitrato redutase e glutamina sintetase e concentração de aminoácidos livres totais na parte aérea e nas raízes dessas gramíneas, quando submetidas a alta e baixa dose de nitrogênio. Na baixa dose de nitrogênio os cultivares Marandu, Xaraés, Piatã, Basilisk e Mombaça mantiveram o número de perfilhos constante e diminuíram o número de folhas e a área foliar, enquanto o cultivar Tanzânia manteve o número de perfilhos e área foliar constantes e reduziu o número de folhas e os cultivares Aruana e Massai aumentaram o número de perfilhos e de folhas e reduziram a área foliar. A baixa dose de nitrogênio resultou em proporções da massa seca da parte aérea e das raízes distintas entre os cultivares quando comparada à alta dose de nitrogênio, favorecendo o crescimento das raízes. Os cultivares estudados modificaram a partição de nitrogênio, nitrato e amônio da parte aérea e raízes, quando em alto e baixo suprimento de nitrogênio. Essas diferenças refletiram na mais alta eficiência de uso do nitrogênio pelos cultivares Mombaça e Tanzânia. A assimilação do nitrato ocorreu principalmente na parte aérea desses capins. Os cultivares Mombaça e Aruana apresentaram elevadas atividades da enzima nitrato redutase nas folhas diagnósticas. A atividade da enzima glutamina sintetase nas folhas diagnósticas foi mais elevada nos cultivares de Panicum. A concentração de amônio nas raízes foi mais alta nos cultivares Piatã e Xaraés, na baixa dose de nitrogênio, e nos capins Aruana e Mombaça, na dose alta de nitrogênio. A concentração de aminoácidos totais nas folhas diagnósticas foi mais elevada nos cultivares de Panicum, quando submetidos à alta dose de nitrogênio, enquanto nas raízes foi maior nos cultivares de Brachiaria, nas duas doses de nitrogênio. / A better understanding of the metabolic, nutritional and production aspects of nitrogen use by grasses are needed to increase nutrient use efficiency and livestock production sustainability. Thus, this study aimed to analyze some forage grasses cultivars used in Brazilian pastures, for: i) morphological and production changes in shoots and roots; II) absorbed nitrogen partition to improve nitrogen use efficiency and III) nitrogen assimilation, evaluated by the variables: dry matter production, number of leaves and tillers, leaf area, concentration and content of total nitrogen, nitrate and ammonium, nitrogen use efficiency, nitrate reductase and glutamine synthetase activities and free amino acids concentration in shoots and roots of these grasses, when supplied with high and low nitrogen rates. At low nitrogen rate, Marandu, Xaraés, Piatã, Basilisk and Mombaça cultivars kept the number of tillers constant and decreased the number of leaves and leaf area, while Tanzânia cultivar kept the number of tillers and leaf area constant and reduced the number of leaves, and Aruana and Massai cultivars increased the number of tillers and leaves and reduced leaf area. The low nitrogen rate resulted in different shoots to roots proportions among cultivars when compared to the higher nitrogen rate, which favored roots growth. The cultivars changed the partition of nitrogen, nitrate and ammonium between roots and shoots, in both nitrogen rates. These differences reflect the higher nitrogen use efficiency by Mombaça and Tanzânia cultivars. The nitrate assimilation occurred mainly in the shoots of these grasses. Mombaça and Aruana cultivars showed high nitrate reductase activity in diagnostic leaves. The glutamine synthetase activity in diagnostic leaves was higher in Panicum cultivars. Ammonium concentration in the roots was high in Piatã and Xaraés, at low nitrogen rate, and Aruana and Mombaça grasses, at high nitrogen rate. The total amino acids concentration in diagnostic leaves was higher in Panicum cultivars than in Brachiaria cultivars, when high nitrogen rate was supplied, whereas Brachiaria cultivars had this concentration high in the roots, at the two nitrogen rates.
47

Aspectos metabólicos, nutricionais e produtivos de cultivares de Brachiaria e Panicum visando eficiência no uso do nitrogênio / Metabolic, nutritional and production aspects of Brachiaria and Panicum cultivars aiming nitrogen use efficiency

Tiago Barreto Garcez 01 October 2013 (has links)
O melhor entendimento sobre os aspectos metabólicos, nutricionais e produtivos das gramíneas forrageiras quanto ao uso do nitrogênio é necessário para aumentar a eficiência de uso do nutriente e para a sustentabilidade da produção pecuária. Com isso, objetivou-se analisar alguns cultivares de gramíneas forrageiras utilizados em pastagens no Brasil, quanto a: I) modificações morfológicas e produtivas da parte aérea e das raízes; II) formas de partição do nitrogênio absorvido para melhorar a eficiência de uso do nitrogênio e III) forma de assimilação do nitrogênio absorvido, sendo avaliados pelas variáveis: produção de massa seca, números de folhas e perfilhos, área foliar, concentração e conteúdos de nitrogênio total, nitrato e amônio, eficiência de uso do nitrogênio, atividades das enzimas nitrato redutase e glutamina sintetase e concentração de aminoácidos livres totais na parte aérea e nas raízes dessas gramíneas, quando submetidas a alta e baixa dose de nitrogênio. Na baixa dose de nitrogênio os cultivares Marandu, Xaraés, Piatã, Basilisk e Mombaça mantiveram o número de perfilhos constante e diminuíram o número de folhas e a área foliar, enquanto o cultivar Tanzânia manteve o número de perfilhos e área foliar constantes e reduziu o número de folhas e os cultivares Aruana e Massai aumentaram o número de perfilhos e de folhas e reduziram a área foliar. A baixa dose de nitrogênio resultou em proporções da massa seca da parte aérea e das raízes distintas entre os cultivares quando comparada à alta dose de nitrogênio, favorecendo o crescimento das raízes. Os cultivares estudados modificaram a partição de nitrogênio, nitrato e amônio da parte aérea e raízes, quando em alto e baixo suprimento de nitrogênio. Essas diferenças refletiram na mais alta eficiência de uso do nitrogênio pelos cultivares Mombaça e Tanzânia. A assimilação do nitrato ocorreu principalmente na parte aérea desses capins. Os cultivares Mombaça e Aruana apresentaram elevadas atividades da enzima nitrato redutase nas folhas diagnósticas. A atividade da enzima glutamina sintetase nas folhas diagnósticas foi mais elevada nos cultivares de Panicum. A concentração de amônio nas raízes foi mais alta nos cultivares Piatã e Xaraés, na baixa dose de nitrogênio, e nos capins Aruana e Mombaça, na dose alta de nitrogênio. A concentração de aminoácidos totais nas folhas diagnósticas foi mais elevada nos cultivares de Panicum, quando submetidos à alta dose de nitrogênio, enquanto nas raízes foi maior nos cultivares de Brachiaria, nas duas doses de nitrogênio. / A better understanding of the metabolic, nutritional and production aspects of nitrogen use by grasses are needed to increase nutrient use efficiency and livestock production sustainability. Thus, this study aimed to analyze some forage grasses cultivars used in Brazilian pastures, for: i) morphological and production changes in shoots and roots; II) absorbed nitrogen partition to improve nitrogen use efficiency and III) nitrogen assimilation, evaluated by the variables: dry matter production, number of leaves and tillers, leaf area, concentration and content of total nitrogen, nitrate and ammonium, nitrogen use efficiency, nitrate reductase and glutamine synthetase activities and free amino acids concentration in shoots and roots of these grasses, when supplied with high and low nitrogen rates. At low nitrogen rate, Marandu, Xaraés, Piatã, Basilisk and Mombaça cultivars kept the number of tillers constant and decreased the number of leaves and leaf area, while Tanzânia cultivar kept the number of tillers and leaf area constant and reduced the number of leaves, and Aruana and Massai cultivars increased the number of tillers and leaves and reduced leaf area. The low nitrogen rate resulted in different shoots to roots proportions among cultivars when compared to the higher nitrogen rate, which favored roots growth. The cultivars changed the partition of nitrogen, nitrate and ammonium between roots and shoots, in both nitrogen rates. These differences reflect the higher nitrogen use efficiency by Mombaça and Tanzânia cultivars. The nitrate assimilation occurred mainly in the shoots of these grasses. Mombaça and Aruana cultivars showed high nitrate reductase activity in diagnostic leaves. The glutamine synthetase activity in diagnostic leaves was higher in Panicum cultivars. Ammonium concentration in the roots was high in Piatã and Xaraés, at low nitrogen rate, and Aruana and Mombaça grasses, at high nitrogen rate. The total amino acids concentration in diagnostic leaves was higher in Panicum cultivars than in Brachiaria cultivars, when high nitrogen rate was supplied, whereas Brachiaria cultivars had this concentration high in the roots, at the two nitrogen rates.
48

Avaliação de parâmetros do metabolismo de carbono e nitrogênio e de respostas ao estresse na associação de trigo com a bactéria Herbaspirillum seropedicae / Evaluation of carbon and nitrogen metabolism parameters and responses to stress in wheat association with bacteria Herbaspirillum seropedicae

Ortolan, Sarah Romani 28 July 2015 (has links)
Made available in DSpace on 2017-07-10T17:37:10Z (GMT). No. of bitstreams: 1 Sarah_Romano_Ortolan.pdf: 975526 bytes, checksum: d72ab61fc7bdc88b6ddbab0eb3a86e42 (MD5) Previous issue date: 2015-07-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Wheat is considered the main cereal diet of the world population, but in recent years has achieved some gain in productivity of this culture despite having increased the use of nitrogen fertilizer. The use of plant growth promoting bacteria such as Herbaspirillum seropedicae SmR1 among others has been studied to obtain development of plants with less use of nitrogen fertilizers. However there is little information relating the effects of this interaction in plant development and grain yield. Objective of this study was to evaluate the carbon and nitrogen metabolism by certain enzymes, metabolites and indices related to the response to infectious stress on the wheat cultivars CD 104 and CD 120 in association with Herbaspirillum seropedicae bacteria. Two experiments were conducted. The experimental design was completely randomized with four replications in a 4x2. The first factor relates to the conditions inoculation with bacteria and/or nitrogen source in coverage are: control without inoculation with bacteria or added nitrogen fertilizer (C); application of nitrogen fertilizer (50 kg.ha-1) 30 days after sowing (N); inoculation with 106 cells of the bacterium H. seropedicae/seed at planting (Hs) and inoculation with bacteria combined with the application of nitrogen fertilizer (Hs + N) and the second factor refers to the phenological stages (tillering and booting). The results indicated that inoculation with H. seropedicae in wheat seeds of cv.s CD 104 and CD 120 in the two growth stages answered in relation to the indices related to stress with the involvement of enzymes of carbon and nitrogen metabolism. However prominent effect was not noticed to promote plant growth of wheat in late development, nor a deleterious effect of the bacterium for inoculation cv. CD 104 under the experimental conditions. For cv. CD 120 the differential effects indicate lower levels of stress and some level of association to positive effect on productivity when combined inoculation of bacteria to nitrogen fertilization. It was concluded that as well as pathogenic and stressors, H. seropedicae able to beneficially associate with wheat also provides similar interference pattern of carbon and nitrogen metabolism and stress levels / O trigo é considerado o principal cereal da dieta da população mundial, entretanto nos últimos anos tem se obtido pouco ganho de produtividade desta cultura apesar de se ter aumentado o uso de fertilizante nitrogenado. O uso de bactérias promotoras do crescimento vegetal, como Herbaspirillum seropedicae SmR1 entre outras, tem sido estudado para se obter desenvolvimento de plantas com menor uso de fertilizantes nitrogenados. Entretanto existem poucas informações que relacionam os efeitos desta interação no desenvolvimento da planta e de produtividade de grãos. Objetivo deste trabalho foi avaliar o metabolismo de carbono e nitrogênio através de algumas enzimas, metabólitos e índices relacionados à resposta ao estresse infeccioso em trigo das cultivares CD 104 e CD 120 em associação com a bactéria Herbaspirillum seropedicae em dois estádios fenológicos. Foram realizados dois experimentos. O delineamento experimental utilizado foi o inteiramente ao acaso, com 4 repetições, em esquema fatorial 4x2. O primeiro fator refere-se às condições de inoculação com bactéria e/ou fertilização nitrogenada em cobertura, sendo: controle, sem inoculação com bactéria ou adição de fertilizante nitrogenado (C); aplicação de fertilizante nitrogenado (50 kg.ha-1) após 30 dias da semeadura (N); inoculação de 106 células da bactéria H. seropedicae/semente na semeadura (Hs) e inoculação com a bactéria combinada com a aplicação de fertilizante nitrogenado (Hs + N) e o segundo fator refere-se aos estádios fenológicos (perfilhamento e emborrachamento). Os resultados indicaram que a inoculação com H. seropedicae em sementes de trigo das cv.s CD 104 e CD 120 nos dois estádios fenológicos responderam em relação aos índices relacionados ao estresse com envolvimento das enzimas do metabolismo de carbono e nitrogênio. Entretanto não foi percebido efeito proeminente de promoção do crescimento vegetal no final do desenvolvimento do trigo, tampouco efeito deletério da inoculação de bactéria para a cv. CD 104, nas condições experimentais. Para a cv. CD 120 os efeitos diferenciais indicam menor nível de estresse e algum nível de associação para efeito positivo na produtividade quando combinada a inoculação da bactéria com a fertilização nitrogenada. Foi possível concluir que assim como para agentes patogênicos e estressantes, a H. seropedicae, capaz de associar beneficamente com trigo também apresenta padrão semelhante de interferência do metabolismo de carbono e nitrogênio e índices de estresse
49

Untersuchungen zum Blatt- und Wurzelmetabolismus sowie zum Phloem- und Xylemtransport in Zusammenhang mit der Stickstoff-Effizienz bei Raps (Brassica napus L.) / Study on nitrogen efficiency of oilseed rape (Brassica napus L.) in relation to the metabolism in leaves and roots and to the transport in phloem and xylem

Zhou, Zewen 02 November 2000 (has links)
No description available.
50

Les neutrophiles ne sont pas résistants aux glucocorticoides

Hirsch, Gaëlle 07 1900 (has links)
Les neutrophiles sont généralement considérés résistants aux glucocorticoïdes. Cependant, peu d’études comparant l’effet de ces drogues sur les neutrophiles et les autres leucocytes sanguins (monocytes, lymphocytes et éosinophiles) ont été rapportées. Dans notre étude, nous avons évalué la réponse aux glucocorticoïdes de ces deux populations cellulaires chez le cheval et l’homme. Les cellules, préalablement isolées du sang de 6 chevaux et 4 sujets humains sains, ont été incubées pendant 5 h en présence de lipopolysaccharide (LPS; 100 ng/mL) seul ou combiné avec de l’hydrocortisone, de la prednisolone ou de la dexaméthasone (10-8M et 10-6M). L’expression d’ARNm pour l’IL-1β, le TNF-α, l’IL-8, la glutamine synthétase et le récepteur α des glucocorticoïdes (GR-α) a été quantifiée par qPCR. Les neutrophiles équins ont également été incubés pendant 20 h en présence de ces 3 glucocorticoïdes et la survie cellulaire a été évaluée par cytométrie de flux et microscopie optique. Nous avons démontré que les glucocorticoïdes inhibaient l’expression des gènes pro-inflammatoires induite par le LPS pour les deux populations cellulaires chez les deux espèces étudiées. L’expression de la glutamine synthétase était également significativement augmentée par les glucocorticoïdes chez les neutrophiles et les autres leucocytes sanguins équins. De manière générale, l’intensité de la réponse aux glucocorticoïdes s’est avérée similaire dans les 2 populations leucocytaires et chez les deux espèces. Les glucocorticoïdes augmentaient également la survie des neutrophiles équins, phénomène également rapporté dans d’autres espèces. Ainsi, les glucococorticoïdes exercent des effets d’intensité comparable sur les neutrophiles et les autres leucocytes sanguins. Nous spéculons que la faible réponse à la corticothérapie observée lors de maladies inflammatoires chroniques neutrophiliques comme l’asthme sévère ou la Maladie Pulmonaire Obstructive Chronique (MPOC) ne s’explique pas par une corticorésistance intrinsèque des neutrophiles. / Neutrophils are generally considered resistant to glucocorticoids compared to other inflammatory cells. However, there are few studies comparing the effects of glucocorticoids in neutrophils and those of other blood leukocytes (monocytes, lymphocytes and eosinophils). In our study, we assessed glucocorticoid-responsiveness in equine and human peripheral blood neutrophils and in neutrophil-depleted leukocytes. Cells were isolated from 6 healthy horses and 4 human healthy subjects. They were incubated for 5 h with or without lipopolysaccharide (LPS; 100 ng/mL) alone or combined with hydrocortisone, prednisolone or dexamethasone (10-8M and 10-6M). IL-1β, TNF-α, IL-8, glutamine synthetase and Glucocorticoid Receptor α (GR-α) mRNA expression was quantified by qPCR. Equine neutrophils were also incubated for 20 h with or without the three glucocorticoids and cell survival was assessed by flow cytometry and light microscopy. We found that glucocorticoids down-regulated LPS-induced pro-inflammatory mRNA expression in both cell populations and species. These drugs also significantly increased glutamine synthetase gene expression in both equine cell populations. The magnitude of glucocorticoid response was generally similar in both cell populations and species. As reported in other species, glucocorticoids significantly increase the survival in equine neutrophils. Based on these results, it appears that glucocorticoids exert effects of similar magnitude on neutrophils and on other blood leukocytes. We speculate that the poor response to glucocorticoids observed in some chronic neutrophilic human diseases such as severe asthma or Chronic Obstructive Pulmonary Disease (COPD) is not explained by an inherent attenuated response of neutrophils to these drugs.

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