O sistema nervoso simpático (SNS) desempenha importante papel sobre o controle da pressão arterial assim como o endotélio, pela liberação de fatores de relaxamento e contração que atuam sobre a modulação do tônus vascular. A hipertensão renovascular (2R-1C) está associada à elevada produção de espécies reativas de oxigênio, hiperatividade do SNS e disfunção endotelial. A hipótese deste trabalho é de que os agonistas adrenérgicos noradrenalina (NOR) e adrenalina (ADR), catecolaminas endógenas, promovam efeito anti-contrátil devido à ativação da eNOS em aorta de ratos 2R-1C. Este estudo teve por objetivo investigar se a ativação de adrenoceptores (AR) com NOR ou ADR leva à maior ativação da eNOS em aortas de ratos 2R-1C do que em 2R e os mecanismos relacionados. Realizamos curvas concentração-efeito para NOR ou ADR, em aortas com ou sem endotélio de ratos 2R e 2R-1C em ausência (controle) ou presença dos antagonistas ?-AR (propranolol), ?2-AR (ioimbina), e inibidor não seletivo da NOS (L-NAME). Por western blot, verificamos a fosforilação do resíduo de ativação da enzima eNOS, Serina1177 (Ser1177), via ativação de ?-AR ou ?-AR, pela NOR ou ADR em aortas com endotélio, de ratos 2R e 2R-1C e se a via PI3K/AKT e o H2O2 estariam envolvidos nesse processo. Avaliamos a produção de NO pelas células endoteliais isoladas de ratos 2R e 2R-1C, por citometria de fluxo. Realizamos a dosagem de NOR e ADR plasmática e tecidual (adrenais) por meio de HPLC. Nos estudos de reatividade vascular avaliamos a potência (pD2) e eficácia (Emax) dos agonistas em induzir contração. O Emax da NOR foi menor na contração de aorta de ratos 2R-1C comparada a 2R, provavelmente devido à maior atividade da eNOS evidenciada pelo efeito do L-NAME em aorta de 2R-1C. A particularidade mais significativa da resposta da NOR é de que em aorta de ratos 2R-1C, a NOR promove a maior fosforilação de Ser1177 via ?-AR, e esta envolve a participação da via PI3K/AKT e do H2O2, não havendo alteração dos níveis plasmáticos e tecidual de NOR entre 2R e 2R-1C. O estímulo com ADR, em aorta de 2R-1C, promoveu aumento da atividade da eNOS, certificada pelo efeito do L-NAME, que pode contribuir para o menor Emax da ADR em 2R-1C do que em 2R. Entretanto, a ADR promoveu maior fosforilação de Ser1177 via ?-AR, em aorta de ratos 2R-1C, e esta não envolve participação da via PI3K/AKT e do H2O2. Os níveis teciduais de ADR foram semelhantes entre 2R e 2R- 1C, mas a concentração plasmática de ADR foi menor em 2R-1C do que em 2R. Não houve diferença na produção de NO pelas células endoteliais entre 2R e 2R-1C. Os resultados obtidos sugerem que a ativação de ?-AR com NOR envolve participação de H2O2 e da via PI3K/AKT para maior ativação da eNOS em aortas de ratos 2R-1C, mecanismo que pode contribuir para o menor Emax da NOR em aorta de 2R-1C. A ADR ao ativar ?-AR leva à maior ativação da eNOS, porém sem participação efetiva de H2O2 e da via PI3K/AKT em aortas de ratos 2R-1C. / The sympathetic nervous system (SNS) plays important role on the arterial pressure control as well the vascular endothelium by relaxing and contractile factors release that modulates the vascular tone. The renovascular hypertension (2K-1C) is related to the increased production of oxygen reactive species, SNS hyperactivity and endothelium dysfunction. The hypothesis of this work is that the adrenoceptor (AR) agonists noradrenaline (NOR) and adrenaline (ADR), the endogenous catecholamine promote anti-contractile effect due to eNOS activation in 2K-1C rat aorta. This study aimed to investigate if AR activation by NOR or ADR leads to the increased activation of eNOS in 2K-1C rat aorta, and the mechanisms activated by these agonists. Concentration-effect curves were constructed for NOR or ADR, in intactendothelium or denuded aortas isolated from 2K-1C and 2K rats in the absence (control) or in the presence of the AR antagonists propranolol (?-AR) or yohimbine (?2-AR), or the non-selective NOS inhibitor, L-NAME. By using western blot, we have veryfied the the effects of activation of ?-AR ou ?-AR and the phosphorylation of NOS activation site Serine1177 (Ser1177) by NOR or ADR in intact endothelium aorta from 2K and 2K-1C and also whether the PI3K/AKT pathway and hydrogen peroxide (H2O2) are related to this phosphorylation. We evaluated by flow cytometry the NO production in the isolated endotelial cells from 2K and 2K-1C. Plasma and tissue (adrenal) levels of NOR and ADR were measured by HPLC. In the vascular reactivity studies, we evaluated the potency (pD2) and efficacy (Emax) of the agonists in inducing contraction. The Emax induced by NOR was lower in 2K-1C than in 2K rat probably due to the higher activity of eNOS as shown by the effect of L-NAME. The most interesting finding was in 2K-1C aorta that NOR increases the Ser1177 phosphorylation via ?-AR activation that involves the signaling trough PI3K/AKT and H2O2. There is no differences in NOR at the plasma and tissue levels between 2K-1C and 2K. ADR activates more eNOS in 2K-1C rat aorta as shown by the effect of LNAME. It could contribute to the lower Emax of ADR in 2K-1C than in 2K. However, ADR increased Ser1177 phosphorylation via ?-AR activation in 2K-1C rat aorta, which does not involve PI3K/AKT and H2O2 pathway. The tissue levels of ADR were not different between 2K-1C and 2K, but the plasma concentration of ADR was lower in 2K-1C than in 2K. There was no difference in the NO production in the endothelial cells from 2K-1C and 2K. Taken together, our results suggest that ?-AR activation by NOR involves H2O2 and PI3K/AKT that activates eNOS in 2K-1C rat aorta that could contribute to the lower contractile effect induced by NOR in 2K-1C. ?-AR activation by ADR leads to the eNOS activation without activation of H2O2 and PI3K/AKT pathway in 2K-1C rat aorta.

Philosophiae Doctor - PhD / Plants used in traditional medicines have served as sources of some of the drug compounds used in medicines today, and could still serve as leads for then development of new drugs to treat existing chronic diseases such as hypertension. This study was aimed at the isolation and identification of a cardio-active compound from L. leonurus, a plant commonly used in traditional medicines in South Africa for the treatment of hypertension and other cardiac problems. The possible mechanisms by which the isolated compound produced its effect on the cardiovascular system were explored using the anaesthetized normotensive rat model.Fractionation of the organic extracts of the leaves led to the isolation of a novel diterpene,(13S)-9 , 13 -epoxylabda-6 (19),15(14)diol dilactone (EDD) whose structure was elucidated using infra red (IR), nuclear magnetic resonance (NMR), mass spectroscopy(MS), and X-ray diffraction analysis. In anaesthetized normotensive male Wistar rats, EDD(0.5 mg/kg – 5.0 mg/kg; IV) produced slight non-significant decreases in systolic pressure(SP), diastolic pressure (DP), and mean arterial pressure (MAP) with the lower (0.5 mg/kg– 2.0 mg/kg) doses, while significant increases in SP, DP and MAP occurred with the higher (3.0 mg/kg – 5.0 mg/kg) doses. All doses of EDD administered also produced significant decreases in heart rate (HR).Prazosin and reserpine pre-treatment abolished the vasoconstrictive effect of EDD,suggesting an indirect vasoconstrictive effect for EDD via the release of catecholamines.Atenolol pre-treatment led to increases in the negative chronotropic effect of EDD, while the positive chronotropic effect of dobutamine was significantly decreased by EDD,suggesting the involvement of the 1 adrenoceptor in the negative chronotropic effect of EDD. In animals pre-treated with verapamil, a cardio-selective Ca2+ channel blocker, no significant changes in HR occurred with all EDD doses, but HR values were significantly lower than those obtained with EDD in non pre-treated animals.The results of this study indicate that (13S)-9 , 13 -epoxylabda-6 (19),15(14)diol dilactone, a novel dilactone diterpene isolated from the leaves of L. leonurus has an effect on the cardiovascular system. EDD exhibits a dual effect on the cardiovascular system by producing a vasoconstrictive effect accompanied by bradycardia. The vasoconstrictive effect of EDD is probably due to the release of catecholamines, while the negative chronotropic effect is probably due to 1 adrenoceptor antagonism. Further studies are however required to fully determine the mechanism by which EDD produces its cardiovascular effects.

This thesis is concerned with a very common disorder of iron homeostasis: iron deficiency. The specific focus is the manner in which iron deficiency influences physiological responses to hypoxia in humans. This work is predicated on observations made over many decades in vitro and in vivo, suggesting that variations in the bioavailability of iron have important consequences for certain biological processes known to depend on oxygen availability. Three separate but related studies together form the basis for this thesis. The first two, Study A and Study B, adopt a similar approach in recruiting healthy volunteers who differ according to iron status, yielding iron-deficient and iron-replete groups in both cases. In Study A, the behaviour of the pulmonary circulation is investigated during a sustained hypoxic exposure, before and after an intravenous infusion of iron. In Study B, skeletal muscle metabolism is explored, both at the level of high-energy phosphate metabolism and the integrated physiological responses to exercise on a cycle ergometer. In the third study, Study C, a different approach is taken, recruiting patients with chronic obstructive pulmonary disease (COPD), and exploring the prevalence and associations of iron deficiency in this condition. Chapters 2 and 3 describe experiments using sustained hypoxia in a normobaric chamber, during which the pulmonary circulation is assessed non-invasively using Doppler echocardiography. These reveal augmented hypoxic pulmonary vasoconstriction (HPV) in iron-deficient individuals, who also exhibit greater sensitivity to the effects of an infusion of intravenous iron. Additionally, the way in which certain circulating mediators important for iron haemostasis change over the course of these hypoxic exposures, and how iron status influences these responses, is explored. Chapter 4 reports the findings of experiments using 31P-magnetic resonance spectroscopy and cardiopulmonary exercise testing, which demonstrate abnormal whole-body metabolism in iron-deficient individuals during large muscle-mass exercise, despite the absence of a clear defect in mitochondrial oxidative phosphorylation. Intravenous iron is found to have significant effects to alter the lactate threshold in healthy individuals, but the effects are more striking in iron-deficient individuals. Collectively, these experiments imply that iron deficiency promotes a more glycolytic phenotype. Chapter 5 explores iron deficiency in COPD, a condition in which pulmonary vascular disease, hypoxia and skeletal muscle dysfunction coexist, and examines some of the difficulties in assessing iron status in the setting of a chronic inflammatory disorder. Iron deficiency is found to be common, and unexpectedly associated with significantly more severe hypoxaemia, in patients with COPD. Possible reasons for these findings, and their clinical implications, are considered. Chapter 6 provides a summary of the main conclusions to be drawn from the studies presented in this thesis.

This thesis is concerned with the role of iron in modulating right ventricular (RV) afterload during exercise in healthy people aged between 50 and 80 years. This is predicated on the requirement of the hypoxia-inducible factor (HIF) pathway for ferrous iron. A secondary objective is to examine the reactive oxygen species (ROS) hypothesis in human hypoxic pulmonary vasoconstriction (HPV) using exposure to hyperoxia. Chapters 3 and 4 describe basal relationships that may affect the HIF pathway and exercise capacity during ageing. These were explored in 113 participants using blood tests and exercise tests. Age and inflammatory factors, C-reactive protein, and ferritin were associated with impaired exercise capacity. In addition, ageing did not significantly affect haematological variables or iron status indicators. Chapters 5 and 6 describe the effect of a single intravenous iron infusion on the haematological variables in 32 participants in a randomised, placebo-controlled and double-blinded study. The effects of iron infusion on RV afterload during light exercise, and exercise capacity during heavy exercise, were examined in these participants. With iron infusion, erythropoietin production, and the increase in RV afterload during light exercise were blunted, potentially indicating involvement of the HIF pathway. However, blunting of RV afterload neither influenced the cardiac output during light exercise nor exercise capacity. Chapter 7 describes a study of 11 healthy volunteers, which investigated the ROS hypothesis in HPV using acute isocapnic hypoxia following an 8-hour exposure to hyperoxia. This sustained hyperoxic exposure did not influence the hypoxic behavior of the pulmonary vasculature. This thesis demonstrates the complex relationship between iron status and exercise capacity in older adults. It shows that the decrease in RV afterload during exercise caused by intravenous iron supplementation does not lead to an augmented cardiac output or exercise capacity. Finally, it calls into question the role of ROS in HPV.

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-02-02T13:02:03Z No. of bitstreams: 1 arquivototal.pdf: 2110108 bytes, checksum: 608a6dd3b7ef97ff56ce7a532de666a1 (MD5) / Made available in DSpace on 2017-02-02T13:02:03Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2110108 bytes, checksum: 608a6dd3b7ef97ff56ce7a532de666a1 (MD5) Previous issue date: 2016-04-26 / This study aimed to quantify the content of phenolic compounds of the bark extracts of the stem and pulp of the fruit of Hymenaea rubriflora Ducke and its effect on vascular reactivity to contractile and relaxing agents thoracic aorta of rats, as well as oxidative stress and biochemical parameters. extraction was performed phenolic compounds of the shell, the stem and the pulp using the ratio of water and alcohol solvents (1: 1). The presence of total phenolic compounds was performed by the Folin-Ciocalteu method and the antioxidant capacity for the kidnapping of free radical DPPH of hydroalcoholic extracts. The vasorelaxant effect of the hydroalcoholic extract of the stem bark (HR-HAc), as well as acute toxicity for the realization of rats supplemented rats with HR-HACwas studied at doses of 25 (G25), 50 (G50) and 150 mg / kg (G100) for four weeks (8 animals / group). Biochemical analysis was used to quantify nitrite (NO), malondialdehyde (MDA), antioxidant activity, lipid profile, and glucose. It was identified that the HR-HAcshowed higher concentration of phenolic compounds (274, 63 mg AC Gál / g) and antioxidant activity (5076.50 micromol Tx / g), the fruit pulp extract (HR-HAp) ( Ac Ga 3.49 mg / g and 29.26 micromol Tx / g, respectively). In addition, the HR-HAcrelaxed the aorta pre-contracted with mouse FEN concentration dependent manner in both, presence and absence of functional endothelium (EC50 = 5.0 ± 1.1 vs. 8.7 ± 1.8 ug / mL), and the attenuated relaxation in the presence of L-NAME (EC50 = 142.4 ± 21.6 mg / mL), however, was not observed change in the relaxation induced by the extract in the presence of indometHAcin (EC50 = 2.9 ± 0.3 ug / mL). Acute supplementation HR-HAcshowed low toxicity, while the chronic supplementation showed increased relaxation potency of ACh compared to G50 G100 (pD2 = 7.8 ± 0.1 vs. 7.6 ± 0.1). The cumulative curve with sodium nitroprusside (SNP) showed greater potency in relaxing G100 compared to G50 (pD2 = 12.1 ± 0.04 vs 11.3 ± 0.3). The contractile efficacy without endothelium in rat aorta was reduced G100 (Emax = 40.4 ± 5.1%). The cumulative curve FEN in the presence of L-NAME showed reduction in contractile potency in rat aorta with endothelium G25 and G50 in comparison GC in the presence and absence of L-NAME (pD2 = 6.4 ± 0.1, 6.4 ± 0.1 and 6.9 ± 0.1 respectively) as well as contractile efficiency of FEN was reduced in the groups G25, G50 and G100 when compared to the control group in the presence and absence of L-NAME (Emax = 61.4 ± 5.2, 74.4 ± 5.3, 76.9 ± 3.7, 100 and 100%, respectively). It was found that there was no significant difference in the values of antioxidant capacity, NO, lipid profile and glucose in plasma of the treated animals and the control group. Thus, it is concluded that this product may be indicated for prophylactic benefits to pathologies related to changes in vascular reactivity. / O presente estudo teve como objetivo quantificar o conteúdo de compostos fenólicos de extratos da casca do caule e da polpa do fruto da Hymenaea rubriflora Ducke e sua ação na reatividade vascular a agentes contráteis e relaxantes de aorta torácica de ratos, assim como extresse oxidativo e parâmetros bioquímicos. Foi realizada extração dos compostos fenólicos da casca do caule e da polpa utilizando como solventes água e álcool etílico (1:1). A presença de compostos fenólicos totais foi realizada pelo método de Folin-Ciocalteau e a capacidade antioxidante pelo sequestro do radical livre DPPH dos extratos hidroalcóolicos. O efeito vasorrelaxante do extrato hidroalcóolico da casca do caule (HR-HAc), assim como a toxicidade aguda para a realização da suplementação de ratos Wistar com o HR-HAc foram investigados nas doses de 25 (G25), 50 (G50) e 150 mg/kg (G100) por quatro semanas (8 animais/grupo). A análise bioquímica foi utilizada para quantificar nitrito (NO), malondialdeído (MDA), atividade antioxidante, perfil lipídico e glicose. Identificou-se que o HR-HAc apresentou maior concentração de compostos fenólicos (274, 63 mg ác gál/g) e atividade antioxidante (5076,50 μmol Tx/g), que o extrato da polpa do fruto (HR-HAp) (3,49 mg ác gál/g e 29,26 μmol Tx/g, respectivamente). Além disso, o HR-HAc relaxou a aorta de rato pré-contraída com FEN de maneira dependente de concentração em ambos, presença e ausência do endotélio funcional (CE50 = 5,0 ± 1,1 vs 8,7 ± 1,8 μg/mL), sendo o relaxamento atenuado na presença do L-NAME (CE50 =142,4 ± 21,6 μg/mL), entretanto, não foi observado alteração no relaxamento induzido pelo extrato na presença da indometacina (CE50 = 2,9 ± 0,3 μg/mL). A suplementação aguda do HR-HAc apresentou baixa toxicidade, enquanto que a suplementação crônica mostrou aumento da potência relaxante da ACh em G50, quando comparado ao G100 (pD2 = 7,8 ± 0,1 vs 7,6± 0,1). A curva cumulativa com nitroprussiato de sódio (NPS) apresentou maior potencia relaxante em G100 quando comparado a G50 (pD2= 12,1 ± 0,04 vs 11,3 ± 0,3). A eficácia contrátil em aorta de rato sem endotélio foi reduzida em G100 (Emax = 40,4 ± 5,1%). A curva cumulativa a FEN na presença de L-NAME mostrou diminuição na potência contrátil em aorta de rato com endotélio em G25 e G50 quando comparado GC na presença e ausência L-NAME (pD2 = 6,4 ± 0,1, 6,4± 0,1 e 6,9 ± 0,1, respectivamente), assim como a eficácia contrátil da FEN foi reduzida nos grupos G25, G50 e G100 quando comparado ao GC na presença e ausência de L-NAME (Emax = 61,4 ± 5,2; 74,4 ± 5,3; 76,9 ± 3,7; 100 e 100%, respectivamente). Verificou-se que não ocorreu diferença significativa para os valores da capacidade antioxidante, NO, perfil lipídico e glicose no plasma dos animais tratados e do grupo controle. Desse modo, conclui-se que este produto poderá ser indicado para benefícios profiláticos a patologias relacionadas à alterações na reatividade vascular.

Made available in DSpace on 2015-05-14T13:00:09Z (GMT). No. of bitstreams: 1 aquivototal.pdf: 6424549 bytes, checksum: 68f0a45898f65b35e3ddee6cfd7df481 (MD5) Previous issue date: 2014-12-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The species Spirulina platensis has been linked to improved vascular function, but the effect of its association with strength training on vascular reactivity, had not yet been investigated. The objective was to verify the effects of strength training and S. platensis on vascular reactivity, biochemical parameters and oxidative stress in aorta of Wistar rats. The animals were divided into eight groups: sedentary (SG) and supplemented with S. platensis at doses of 50 (SG50); 150 (SG150) and 500 (SG500) mg/kg and trained groups (TG) and supplemented with S. platensis at doses of 50 (TG50); 150 (TG150) and 500 (TG500) mg/kg. The TG performed strength training for 8 weeks. Biochemical analysis was used to quantify the activity of creatine kinase (CK) and lactate dehydrogenase (LDH) levels of reactive C protein (RCP), nitrite, malondialdehyde (MDA) and antioxidant activity. To evaluate the vascular response induced is a cumulative concentration-response curve to phenylephrine (PHE) (10-9-10-3 M) and acetylcholine (ACh) (10-11-10-4 M). To assess the involvement of nitric oxide used is L-NAME and then a cumulative concentration-response curve to the FEN (10-11-10-3 M) was induced. It has been shown so training reduced by around 30% of time jumps without increasing the activity of LDH, CK, CRP and MDA in any of TG when compared to its control group. The functional experiments, it was observed that for contractile reactivity between the SG groups, only SG500 (pD2 = 5.6 ± 0.04 vs. 6.1 ± 0.06, 6.2 ± 0.02, and 6.2 ± 0.04), a significant reduction in the contractile power to FEN. However GT150 and GT500 for groups reduced the contractile power to FEN (pD2 = 5.0 ± 0.06 and 5.3 ± 0.05 vs. 5.6 ± 0.07 and 5.5 ± 0.05). Being that a dose of 500 mg/kg in the trained group reduced the contractile power above the sedentary. It was also found that the ability of strength training and S. platensis in reducing the contractile response was eliminated in the absence of the endothelium and the presence of L-NAME provided greater contractile response to FEN in all groups, and this enhanced response in TG and SG groups. Regarding the relaxing activity in the SG150 groups (pD2 = 7.0 ± 0.08 vs. 6.4 ± 0.06) and SG500 (pD2 = 7.3 ± 0.02 vs. 6.4 ± 0.06) the curve was shifted to the left, similarly the TG150 groups (pD2 = 7.6 ± 0.08 vs. 7.3 ± 0.02) and TG500 (pD2 = 8.0 ± 0.04 vs. 7.3 ± 0.02) also showed the same behavior. However, supplementation at doses of 150 and 500 mg/kg with training further increased relaxant response to ACh. The role of NO was increased when there was significant increase in nitrite production in the aorta of TG150 and TG500. The production of MDA was reduced in all groups around 80%, while the oxidation inhibition percentage was increased by around 50%, mainly from the TG and SG at doses of 150 and 500 mg/kg indicating an enhancement in action antioxidant between such an association. Thus, the present study demonstrated that chronic supplementation of S. platensis at doses of 150 mg/kg and 500 mg/kg or a strength training program alone are effective in promoting an increase in reactivity to ACh relaxing and a decrease in reactivity FEN contractile in the rat aorta, and that the combination of these two procedures enhances the effects. These effects are accompanied by an increase in nitrite concentration, indicating involvement of the NO pathway, by a reduction of oxidative stress and increased antioxidant activity. / A espécie Spirulina platensis tem sido relacionada à melhoria na função vascular, porém o efeito da sua associação com o treinamento de força sobre a reatividade vascular, ainda não tinha sido investigado. Assim, objetivou-se verificar os efeitos do treinamento de força e da S. platensis sobre a reatividade vascular, parâmetros bioquímicos e o estresse oxidativo, em aorta de ratos Wistar. Os animais foram divididos em oito grupos: sedentário (GS) e suplementados com S. platensis nas doses de 50 (GS50); 150 (GS150) e 500 mg/kg (GS500) e grupos treinados (GT) e suplementados com S. platensis nas doses de 50 (GT50); 150 (GT150) e 500 mg/kg (GT500). Os grupos GT realizaram treinamento de força durante 8 semanas. A análise bioquímica foi utilizada para quantificar a atividade da creatina quinase (CK) e lactato desidrogenase (LDH), níveis da proteína C reativa (PCR), de nitrito, malondialdeído (MDA) e atividade antioxidante. Para avaliar a reatividade vascular induziu-se uma curva concentração-resposta cumulativa à fenilefrina (FEN) (10-9-10-3 M) e à acetilcolina (ACh) (10-11-10-4 M). Para avaliar a participação do óxido nítrico, utilizou-se o L-NAME e em seguida, uma curva concentração-resposta cumulativa à FEN (10-11-10-3 M) era induzida. Foi demonstrado assim que o treinamento reduziu em torno de 30% o tempo de execução dos saltos, sem aumentar a atividade da LDH, CK, PCR e MDA em nenhum dos grupos GT quando comparado com o seu grupo controle. Com relação aos experimentos funcionais, observou-se que para a reatividade contrátil, entre os grupos GS, apenas GS500 (pD2 = 5,6 ± 0,04 vs. 6,1 ± 0,06; 6,2 ± 0,02 e 6,2 ± 0,04), apresentou redução significativa na potência contrátil à FEN. No entanto para os grupos GT150 e GT500 reduziram a potência contrátil à FEN (pD2 = 5,0 ± 0,06 e 5,3 ± 0,05 vs. 5,6 ± 0,07 e 5,5 ± 0,05). Sendo que a dose de 500 mg/kg no grupo treinado reduziu a potência contrátil superior ao sedentário. Verificou-se ainda que a capacidade do treinamento de força e da S. platensis em reduzir a resposta contrátil foi eliminada na ausência do endotélio e a presença do L-NAME proporcionou uma maior resposta contrátil à FEN em todos os grupos, sendo essa resposta potencializada nos grupos GT e GS. Em relação à atividade relaxante nos grupos GS150 (pD2 = 7,0 ± 0,08 vs. 6,4 ± 0,06) e GS500 (pD2 = 7,3 ± 0,02 vs. 6,4 ± 0,06) a curva foi desviada para a esquerda, similarmente os grupos GT150 (pD2 = 7,6 ± 0,08 vs. 7,3 ± 0,02) e GT500 (pD2 = 8,0 ± 0,04 vs. 7,3 ± 0,02) também apresentaram o mesmo comportamento. No entanto, as suplementações nas doses de 150 e 500 mg/kg com o treinamento aumentaram ainda mais a resposta relaxante à ACh. A participação do NO foi reforçada quando se verificou aumento significativo na produção de nitrito na aorta de GT150 e GT500. A produção de MDA foi reduzida em todos os grupos em torno de 80%, enquanto o percentual de inibição da oxidação foi aumentado em torno de 50%, principalmente dos GT e GS nas doses de 150 e 500 mg/kg indicando uma potencialização na ação antioxidante entre essa associação. Assim, o presente estudo demonstrou que a suplementação crônica de S. platensis nas doses de 150 mg/kg e 500 mg/kg ou um programa de treinamento de força são isoladamente eficazes em promover um aumento na reatividade relaxante à ACh e uma diminuição na reatividade contrátil à FEN na aorta de rato, e ainda que a combinação destes dois procedimentos potencializa os efeitos. Estes efeitos são acompanhados por um aumento na concentração de nitrito, indicando participação da via do NO, por uma redução do estresse oxidativo e aumento na atividade antioxidante.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal. / Il est maintenant bien établi que les GHRPs constituent une classe de peptides synthétiques stimulant la sécrétion de l'hormone de croissance par interaction avec leur récepteur spécifique au niveau hypophysaire. Ce récepteur constitué de 364 acides aminés appartient à la famille des récepteurs à sept passages transmembranaires couplé aux protéines G. Il est exprimé non seulement au niveau de l'hypophyse mais également au niveau de l'hypothalamus, des noyaux arqués et paraventriculaires. Récemment, il a été découvert par étude de photomarquage covalent que l'hexarelin, un hexapeptide de la famille des GHRPs se lie de façon spécifique à une glycoprotéine membranaire de poids moléculaire de 84 kDa exprimée dans les membranes de coeur de rat. La liaison de l'hexarelin à ce récepteur au niveau cardiaque induit une forte vasoconstriction coronarienne de façon dose­dépendante dans le modèle de coeur de rat perfusé. Les buts de la présente étude sont premièrement de déterminer si la liaison est spécifique seulement à l'hexarelin ou à d'autres analogues peptidiques de la famille des GHRPs, notamment le dérivé EP 70905. Dans un deuxième temps nous déterminerons si les analogues se liant au récepteur cardiaque des GHRPs exercent un effet vasoconstricteur sur les coronaires utilisant le modèle de cœur de rat perfusé. Troisièmement nous caractériserons les voies de signalisation impliquées dans la vasoconstriction induite par les GHRPs. Les études de compétition entre le radioligand photoactivable [125] I-Tyr-Ala-Bpa­hexarelin et le EP 70905 effectuées sur les membranes de cœur de rat démontrent une affinité de liaison comparable du EP 70905 et de l'hexarelin au récepteur cardiaque des GHRPs. Cependant, le EP 70905 induit une augmentation de la pression coronarienne significativement plus élevée que celle induite par l'hexarelin. Les études des voies de signalisation démontrent que les tyrosine kinases ne semblent pas être impliquées dans la vasoconstriction induite par le EP 70905, puisque la perfusion simultanée avec la génistéine n'affecte pas la vasoconstriction induite par ce peptide. Le rôle de l'endothéline endogène dont la libération est stimulée par le EP 70905 semble être mineur dans la vasoconstriction observée considérant les taux femtomolaires d'endothéline libérés dans le perfusat. De plus, la perfusion simultanée d'un antagoniste des récepteurs d'endothéline, le LU 302872 affecte que faiblement la vasoconstriction induite par le EP 70905. Par contre, la vasoconstriction coronarienne induite par le EP 70905 est inhibée de façon majeure suite à la perfusion de la nifédipine, un bloqueur des canaux calciques de type L. La perfusion simultanée d'un inhibiteur de la protéine kinase C, la chélérythrine affecte de façon significative la vasoconstriction induite par le EP 70905.

Objetivo: checar se poluentes do ar urbano de São Paulo induzem alterações sangüíneas e cardiopulmonares. MM: camundongos Balb/c foram expostos por 7, 14, 21, 30 e 45 dias em 3 câmaras: Limpa (controle), Intermediária (seletiva ao PM) e Suja (ar externo). Após exposição, os animais foram ventilados (FlexiVent) e coletados dados de mecânica pulmonar, sangue, coração e pulmão. Foram registrados PM, CO, SO2 e NO2 diários. Resultados: aos 21 e 45 dias coincidentes com picos de poluição houve aumento da resistência de via aérea (45d p=0,012), leucocitose (21d p < 0,001 e 45d p=0,039) e vasoconstricção pulmonar (21d p=0,034) nos animais da Câmara Suja, sem alteração de coronárias. Nenhum poluente excedeu limites de qualidade de ar / Objective: verify if air pollution of São Paulo city induces alterations in blood and cardiopulmonary systems. MM: Balb/c mice were exposed during 7, 14, 21, 30 and 45 days to 3 chambers: Clean (control), Intermediate (PM only) and Dirty (external air). After exposure, animals were ventilated (FlexiVent) and collected lung mechanics data and blood, heart and lung. PM, CO, SO2 e NO2 were measured daily. Results: on day 21 and 45, coincidently to peak of pollutions, there was proximal airway resistance increase (45d p=0.012), leukocytosis (21d p < 0.001 and 45d p=0.039) and vasoconstriction of peribronchiolar arterioles (21d p=0.034) in animals of Dirty Chamber with no alterations of coronaries. Neither pollutants exceeded the standard limits

Einleitung: Das ARDS (acute respiratory distress syndrome) stellt mit einer schweren Hypoxämie aufgrund eines hohen pulmonalen Recht-Linksshunts, mit einer pulmonalen Hypertonie und einer diffusen inflammatorischen Reaktion der gesamten Lunge trotz einiger etablierter Therapieansätze ein häufig letal endendes Krankheitsbild dar. Neben zahlreichen neuen klinisch-experimentellen Therapieansätzen erlangt die i.v.-Aplikation von Almitrindimesilat größere Beachtung. Zuerst verabreicht bei chronisch obstruktiven Atemwegserkrankungen erkannte man bald, daß neben der ventilationssteigernden Wirkung auch Effekte an der pulmonalen Gefäßstrombahn im Sinne einer Optimierung des Ventilations-Perfusions-Verhältnisses für die zu beobachtende Verbesserung der pulmonalen Oxygenation verantwortlich sind. Eine Potentierung der Hypoxischen Pulmonalen Vasokonstriktion (HPV) wird dafür verantwortlich gemacht. Kontrovers diskutierte Dosierungen in Tierversuchen und vereinzelten Anwendungen am Menschen legten nahe, daß systemische Dosis-Wirkungsanalysen erfolgen mußten. Desweiteren sollte untersucht werden ob die im Rahmen des ARDS zu beobachtende Pulmonalarterielle Hypertonie durch das Medikament zunimmt. Methoden: Mit Hilfe eines durch repetitive saline bronchoalveoläre Lavage induzierten ARDS-Modells am Schwein wurden an 14 Schweinen in Narkose prospektiv randomisiert entweder sechs ansteigende Dosen (0,5, 1, 2, 4, 8, und 16 (g/kgKG/min) Almitrindimesilat oder dessen Solvens Apfelsäure appliziert. Während des Versuches wurde volumenkontrolliert beatmet (FiO2 1,0 , PEEP 5 cm H2O). Nach je 30 minütiger Infusion der entsprechenden Dosierungen wurde Parameter des Gasaustausches und der pulmonalen sowie der systemischen Hämodynamik erhoben. Innerhalb der Gruppen wurde mittels des einfachen ANOVA-Test mit Bonferonikorrekturfaktor, zwischen den Gruppen mit dem Mann-Whitney-Test auf signifikante Unterschiede hin untersucht. Signifikanz der Unterschiede wurde bei einer Irrtumswahrscheinlichkeit p ( 0,05 angenommen. Ergebnisse: Niedrige Dosierungen, 0,5 - 2 (g/kgKG/min, reduzieren die venöse Beimischung und führen zu einem statistisch signifikanten Anstieg des Sauerstoffpartialdruckes mit einem Optimum für diesen günstigen Einfluß bei einer Infusionsrate von 1 µg/kgKG/ min. Hohe Dosierungen hingegen erhöhen die venöse Beimischung und senken den Sauerstoffpartialdruck und führen damit zu einer erheblichen Verschlechterung des Krankheitsbildes im Vergleich zu niedrigen Dosen und der Ausgangssituation bzw. dem nichttherapierten Tieren. Der Pulmonalarterielle Mitteldruck nahm nach Almitrindimesilatgabe geringfügig zu. Mit zunehmender Dosierung erreichte der MPAP höhere Werte als zum Zeitpunkt des stabilen Lungenschadens, jedoch waren keine dieser Steigerungen statistisch signifikant. Diskussion: Almitrindimesilat zeigte einen dosisabhängigen Einfluß auf den pulmonalen Gasaustausch. Dieser wirkt sich in niedrigen Dosierungen positiv auf die Hypoxämie aus. Der Pulmonalarterielle Druck bleibt konstant. Als Mechanismus der positiven Auswirkung liegt eine Potenzierung der HPV für niedrige Dosierungen nahe. / Objectives: The ARDS (acute respiratory distress syndrome) is characterized by a diffuse inflammatory response, a pulmonary hypertension and a severe hypoxemia mainly because of ventilation-perfusion-mismatching. Despite of some new therapeutical strageties the mortality remains high. Almitrine bismesylate was first described to improve arterial oxygenation in COPD- patients primary by increasing alveolar ventilation due to a stimulating effect on peripheral chemoreceptors. Furthermore several experimental studies have confirmed that almitrine improves PaO2 possibly because of reducing the ventilation- perfusion-mismatching by enhancing the hypoxic pulmonary vasoconstriction (HPV). Controversal results have been reported concernig the doses of intravenous Almitrine to be beneficial. Furthermore it remains unclear to what extend the pulmonary hypertension is augmented by a drug which is inhancing or restroring the HPV. Therefore we determined a dose-response-curve of almitrine in a animal model of ARDS. Methods: After induction of anaesthesia the ARDS-Modell was induced by repeated saline wash-0,5-2 ?g/kgKG/minout of surfactant. 14 swine received almitrine or the solvent in a prospective randomized manner. During the experiment the volume controlled mode of mechanical ventilation remained unchanged (FiO2 1,0 , PEEP 5 cm H2O). After the increasing doses of 0,5, 1, 2, 4, 8 and 16 ?g/kgKG/min each for 30 min we investigated the effects of i.v. almitrine on the pulmonary gasexchange and the pulomanry and systemic hemodynamics. Statistics were carried out by Two way Analysis of Variance p