O selênio (Se) é um micronutriente importante na dieta de mamíferos e tem sido descrito com importante papel na função imune. É constituinte de mais de 25 selenoproteínas na forma do aminoácido selenocisteína, sendo este elemento crítico na manutenção do sistema de defesa antioxidante. Uma dieta complementar com Se pode ser benéfica no tratamento de doenças correlacionadas com altos níveis de estresse oxidativo, como a doença de Chagas, enfermidade negligenciada causada por Trypanosoma cruzi. O objetivo deste estudo foi avaliar os efeitos do Se em ratas Wistar prenhas infectadas pela cepa Y de T. cruzi. O tratamento com Se desencadeou aumento no peso e comprimento fetal, bem como no diâmetro e peso placentário. Também foi observada diminuição da parasitemia. Não ocorreram alterações significativas nas concentrações de NO e no número de ninhos de amastigotas no coração. A avaliação histológica das placentas mostrou elevado número de ninhos de amastigotas nos animais do grupo infectado e tratado. A redução da concentração de citocinas pró-inflamatórias e de populações de células T desencadeou uma resposta voltada ao padrão Th-2, característico da gestação, fato que provavelmente contribuiu no aumento do parasitismo placentário encontrado nos animais tratados com Se. Assim, é possível que a administração de Se, durante a prenhez, poderia alterar a resposta imune placentária local, favorecendo a instalação do parasita. Mais estudos são necessários para avaliar a interação entre o Se e a doença de Chagas durante a prenhez. / The selenium (Se) is an essential micronutrient in the diet ofmammals and has an important role in the immune function. A range of 25 selenoproteinshas Sein its structure and most of them in the form of amino acid selenocysteine, being this element involved in the in maintenance of the antioxidant defense. Diet with Se is beneficial in the treatment of diseases correlated with high levels of oxidative stress, like Chagas\' disease, a neglected illness caused by Trypanosoma cruzi. The objective of this study was to evaluate the effects of selenium in the immune response of pregnant Wistar rats infected withtheY strain of T. cruzi. Se treatment triggered enhanced fetal weight and length and placental diameter and weight. It was observed decreased parasitemia. No significant alterations in NO concentrations and amastigote nests in heart were observed. The histological evaluation of placenta displayed an enhanced number of amastigote nests in infected and Se treated animals. The reduction of pro-inflammatory cytokines and T cell populations triggered a Th-2 immune response, which is the hallmark of the gestation period. This fact probably led to the raise in parasite nests in placenta of infected and Se treated animals. So it is possible that the Se supplementation during pregnancy could impair the local placental immune response. Further studies are needed to assess the interaction between selenium and the acute Chagas\' disease during pregnancy.

A implantação embrionária determina eventos biológicos de fundamental importância para o desenvolvimento do embrião e para o sucesso da gestação. O fator de inibição de migração de macrófagos (MIF) é uma das muitas citocinas que atuam durante a gestação, desempenhando múltiplas funções biológicas e atividades pró-inflamatórias, em resposta a infecções ou à presença de toxinas bacterianas e estresse. Este estudo investigou a expressão de MIF na interface materno-placentária em condições infecto-inflamatórias simuladas no organismo materno pela administração de LPS de Escherichia coli, período imediatamente após a implantação embrionária. Na primeira etapa foi administrado LPS nas doses de 0,06, 0,1, 0,2 e 0,3 <font face=\"Symbol\">mg/g de peso corporal aos 7,5 dias de gestação e o perfil gestacional foi analisado. Na segunda etapa a dose mais adequada (de 0,1 <font face=\"Symbol\">mg/g) foi administrada e a gestação interrompida 30 minutos, 1, 3 e 6 após. Com esta dose observou-se diminuição o padrão de expressão protéica de MIF durante as 6 horas seguintes ao estímulo com LPS, enquanto que a expressão gênica permaneceu estável, aumentando significativamente apenas após 6 horas de tratamento. / The embryo implantation determines biological events essential for embryo development and the success of pregnancy. The macrophage migration inhibitory factor (MIF) is one of many cytokines that act during pregnancy, playing multiple biological functions and pro-inflammatory activities in response to infection or the presence of bacterial toxins and stress. This study investigated the expression of MIF in the maternal-placental interface in infectious and inflammatory conditions simulated in the mother by the administration of LPS from Escherichia coli, on the period immediately after embryo implantation. In the first step LPS was administered at doses of 0.06, 0.1, 0.2 and 0.3 <font face=\"symbol\">mg / g body weight at 7.5 day gestation and pregnancy profile was analyzed. In the second step the more appropriate dose (0.1 <font face=\"symbol\">mg / g) was administered 30 minutes and stopped pregnancy, 1, 3 and 6 after. At this dose there was a decrease in the pattern of protein expression of MIF during the 6 hours of stimulation with LPS, while the gene expression remained stable, significantly increasing only after 6 hours of treatment.

Perdas gestacionais, malformações, restrição de crescimento intrauterino (IUGR) são associadas a gestações diabéticas. Para ampliar o conhecimento nesse tema, nosso grupo desenvolveu um modelo de gestação complicada por diabetes tipo 1 em camundongos que, nessa tese, foi utilizado para analisar o ciclo estral, desenvolvimento fetal e organização placentária. O diabetes foi induzido por aloxana e estudado em dois períodos 30-50D (curto prazo) e 90-110D (longo prazo). Placentas e fetos foram coletados, pesados, e submetidos a técnicas moleculares, bioquímicas e morfológicas. Detectaram-se alterações no perfil temporal do ciclo estral. O grupo 30-50D apresentou altas taxas de perdas embrionárias e IUGR, e o 90-110D malformações, mortes fetais, IUGR e aumento no peso placentário. As placentas diabéticas apresentaram aumento e desorganização da zona juncional, redução do labirinto e vasodilatação. A expressão dos colágenos I e III aumentou e a do V diminuiu em 30-50D, porém, a deposição destes aumentou concomitante com a redução da atividade da MMP9. A deposição dos colágenos III e V e a atividade da MMP2 aumentaram em 90-110D. Nossos resultados reiteram a importância do fator temporal nas complicações do diabetes sobre a gestação. / Gestational loss, malformations and intrauterine growth restriction (IUGR) are often associated with pregnancies. To increase the knowledge about this topic, our group has developed a model of pregnancy complicated by type 1 diabetes in mice. In this study, was analyzed the estrous cycle and the fetal and placental development. For this, diabetes was induced by alloxan and studied in two time-periods 30-50D (short term) and 90-110D (long term). Placentas and fetuses were collected, weighed and analyzed by biochemical, morphological and molecular procedures. We detected changes in the temporal profile of the estrous cycle. The 30-50D group showed high rates of embryonic loss and IUGR whereas malformations, fetal death, IUGR and increased placental weight was detected in 90-110D. Increase and disorganization of junctional zone, reducing labirinth and vasodilation characterize diabetic placentas. The expression of collagen I and III was increased whereas collagen V decreased in the 30-50D. The deposition of this collagen, however increased concomitant with the reduction of MMP9 activity. In 90-110D deposition of collagen III and V and the MMP2 activity was increased. Together, our results reinforce the relevance of the time factor in the complications of diabetes on pregnancy.

Nos dias atuais, a utilização de fitoterápicos tem crescido acentuadamente. No Brasil, um país cuja flora nativa é riquíssima, tem-se investido substancialmente em pesquisas nesta área. Isto se deve, em parte, à necessidade de novos medicamentos, ao interesse na comercialização destes produtos, ao interesse na preservação da cultura popular e da reserva da flora nacional. Paralelamente a este cenário, está a crença de que medicamentos fitoterápicos são inofensivos em circunstâncias especiais tais como: gravidez, hipertensão, diabetes, etc. É como se os fitoterápicos atuassem especificamente sobre uma determinada patologia não sobre o metabolismo como um todo. A Mikania glomerata Sprengel, conhecida popularmente como guaco e originária da América do Sul, é uma planta subarbustiva, que nasce nas matas e cerrados e, que se adapta muito bem ao cultivo doméstico. Ela é vastamente utilizada pela população no tratamento de doenças como a asma, bronquite, e reumatismo, além de possuir efeito antifúngico, antimicrobiano, antialérgico, antiinflamatório e antiofídico, na grande maioria das vezes administrada sem supervisão de profissionais da área da saúde. Neste estudo, nosso objetivo é estudar a possível ação do extrato vegetal de Mikania glomerata Sprengel (guaco) no perfil reprodutivo e gestacional de camundongos (Mus musculus domesticus) e determinar se a administração desta droga pode comprometer o embrião/feto e placenta durante a prenhez. Este estudo mostrou que a utilização do extrato de Mikania glomerata em doses supra-terapêuticas pode atuar sobre processos morfofuncionais orgânicos, interferindo no crescimento placentário e fetal e podendo levar ao insucesso gestacional e ao aparecimento de defeitos congênitos. Além disto, a diminuição do crescimento fetal observado nas doses consideradas terapêuticas também é um alerta para o uso inadvertido do extrato de guaco sem acompanhamento médico devido, em qualquer período gestacional. / Nowadays, the use of medicinal plants has grown dramatically. In Brazil, a country whose native flora is rich, has invested substantially in research in this area. This is due in part to the need for new drugs, the interest in marketing these products, the interest in the preservation of popular culture and the reserve of national flora. In parallel with this scenario is the belief that phitotherapeutics are harmless in special circumstances such as pregnancy, hypertension, diabetes, etc. It is as if the herbal acted specifically in a determinate disease not on the metabolism as a whole. The Mikania glomerata Sprengel, popularly known as guaco and originating from South America, is a plant undergrowth, which rises in the forests and savannahs, and that lends itself very well to domestic cultivation. It is widely used by local people in the treatment of diseases such as asthma, bronchitis, and rheumatism, as well as having antifungal effect, antimicrobial, antiallergic, anti-inflammatory and anti-snakebite, in most cases administered without the supervision of health professionals. In this study, our goal is to study the possible action of plant extract of Mikania glomerata Sprengel (guaco) in pregnancy and reproductive profile of mice (Mus musculus domesticus) and determine whether the administration of this drug may affect the embryo / fetus and placenta during pregnancy . This study showed that the use of extract of Mikania glomerata at supratherapeutic doses can act on morphofunctional organic processes, interfering with fetal and placental growth and could lead to pregnancy failure as well as contribute to the appearance of birth defects. Furthermore, the decrease in fetal growth observed in therapeutic doses is also considered a warning to the inadvertent use of the extract guaco without medical supervision because, at any gestational period.

O mecanismo de imunorregulação gerado pelo catabolismo do triptofano pela IDO protege o feto contra a resposta imunológica materna. Neste estudo, esse mecanismo foi em avaliado em gestantes imunossuprimidas e portadoras de transplante renal. Examinou-se a expressão da IDO e sua atividade nos compartimentos placentários de gestantes saudáveis e portadoras de transplante. Células produtoras de IDO e IFN-<font face=\"Symbol\">g foram imunolocalizadas na região vilosa e região decidual em ambos os grupos analisados, com mudanças no tipo celular envolvido nestas expressões nas gestantes transplantadas. Os níveis de IDO e sua atividade, assim como seus fatores de regulação NF-kB, IFN-<font face=\"Symbol\">g e IL-10 estavam diminuídos na região vilosa. No compartimento decidual a atividade enzimática da IDO estava aumentada nas gestantes transplantadas, mas não dos seus reguladores. Sendo assim, eixo de imunorregulação gerado por IDO-IFN-<font face=\"Symbol\">g na interface placentária de gestantes portadoras de transplante renal responde diferencialmente a insultos ocasionados pela utilização de imunossupressores durante a gestação. / The mechanism of immunoregulation generated by the catabolism of tryptophan by IDO protects the fetus against maternal immune response. In this study, this mechanism was evaluated in renal transplanted pregnant women and immunosuppressed. We examined the expression and activity of IDO in placental compartments of healthy pregnant women and patients with transplants. IDO and IFN-<font face=\"Symbol\">g producing cells were imunolocalizated in villous and decidual region in both groups analyzed, with changes in cell type involved in these expressions in pregnant patients transplanted. The levels and activity of IDO, as well as their regulatory factors NF-kB, IFN-<font face=\"Symbol\">g and IL-10 were decreased in the villous region. In the decidual compartment IDO activity was increased in pregnant women transplanted, but not their regulators. Thus, the axis of immunoregulation generated by IDO and IFN-<font face=\"Symbol\">g in placental interface of pregnant women with renal transplantation responds differently to insults caused by the use of immunosuppressive drugs during pregnancy.

Le développement normal du placenta est la clé du succès de la grossesse. La croissance trophoblastique et vasculaire est une composante cruciale du développement placentaire. Un déficit dans ces processus conduit à des complications de la grossesse, telles que la Toxémie Gravidique (TG) ou le Retard de Croissance Intra-Utérin (RCIU). Récemment, un nouveau facteur angiogène spécifique des glandes endocrines vient d'être découvert : EG-VEGF pour « Endocrine Gland-derived Vascular Endothelial Growth Factor» ou PROK1 pour Prokineticin 1, facteur qui se lie à deux récepteurs couplés aux protéines G, PROKR1 et PROKR2. Dans des publications récentes du laboratoire, l'équipe a montré que I) EG-VEGF est fortement exprimé dans le placenta, II) son expression est majoritaire au 1er trimestre et III) les niveaux circulants d'EG-VEGF sont augmentés dans la TG, pathologie qui résulte de défauts vasculaires importants. L'ensemble de ces résultats convergeait vers un effet potentiel important d'EG-VEGF sur le placenta. Les objectifs de ma thèse ont été de caractériser (1) l'effet angiogène d'EG-VEGF sur les cellules endothéliales microvasculaires placentaires (HPEC), (2) d'étudier sa régulation par l'hCG (human Chorionic Gonadotropin), et (3) d'étudier son expression au 3ème trimestre dans les grossesses avec RCIU. J'ai montré que 1) EG-VEGF est un facteur angiogène important, et que ses effets angiogènes sont médiés par PROKR1, alors que c'est PROKR2 qui est impliqué dans ses effets sur la perméabilité, 2) l'hCG augmente la sécrétion d'EG-VEGF, ainsi que l'expression des PROKRs au 1er trimestre de la grossesse, et 3) EG-VEGF augmente la prolifération et la survie des trophoblastes, et est augmenté ainsi que ses récepteurs au 3ème trimestre dans les grossesses avec RCIU. L'ensemble de ces résultats montre qu'EG-VEGF est un nouveau facteur de croissance du placenta. Sa dérégulation dans la TG et le RCIU suggère qu'il pourrait servir de marqueur précoce prédictif pour ces pathologies et être utilisé comme cible thérapeutique potentielle dans l'avenir. / Growth of the placental villi is a key event in placental development during human pregnancy. Many growth factors have been shown to control this process; however their ubiquitous expression makes them less specific to this tissue. Recently, a new growth factor named EG-VEGF (Endocrine Gland-derived Vascular Endothelial Growth Factor), specific to endocrine glands including the placenta, has been identified. It mediates its biological activity via two G protein coupled receptors, Prokineticin Receptor 1 and 2 (PROKR1 and PROKR2). In recent work from our laboratory, we have shown that I) EG-VEGF is abundantly expressed in human placenta II) EG-VEGF levels are highest during the first trimester and III) EGVEGF circulating levels are increased in preeclampsia (PE). Altogether, our results suggested that EG-VEGF might be an important regulatory factor in the placenta. My aim was to investigate (1) EG-VEGF angiogenic effects on microvascular cells within the placenta (HPEC), (2) its hormonal regulation with hCG (human Chorionic Gonadotropin), and (3) its expression in IUGR (Intrauterine Growth Restriction) pregnancies in the third trimester. Our results show that 1) EG-VEGF increases angiogenic processes in HPEC cells via PROKR1, and also their permeability via PROKR2 2) hCG increases EG-VEGF secretion and PROKR expression in the first trimester placenta and 3) EG-VEGF increases both trophoblast proliferation and survival, and its expression is dysregulated in IUGR in the third trimester of pregnancy. Altogether, our results show that EG-VEGF is a new placental growth factor. Its dysregulation in PE and IUGR suggests that it can be considered as a potential diagnostic marker and a potential therapeutic target in future.

Les syncytines sont des gènes d'enveloppes rétrovirales (env) capturés qui sont essentiels pour l'établissement du placenta chez les mammifères. Il a été proposé que la diversité des syncytines capturées explique pourquoi le placenta est l'organe le plus variable chez les mammifères. Ici nous avons employé deux approches pour étudier le lien entre la capture d'env et l'émergence et la diversité des structures placentaires. D'abord, nous avons étudié la placentation des Hyaenidae, les seuls carnivores à présenter un placenta très invasif hémochorial, comme l'humain. Comme tous les carnivores, les hyènes expriment la syncytin-Car1 précédemment décrite, mais nous avons identifié une nouvelle env, capturée uniquement chez ces dernières, que nous avons nommée Hyena-Env2. Ce nouveau gène est présent au même locus chez toutes les hyènes, ayant été capturé pendant la radiation de la famille. Il est non-fusiogène mais a néanmoins été conservé pendant plus de 10 millions d'années et est exprimé à l'interface materno-fœtale du placenta, ce qui en fait un gène candidat pour expliquer le passage à la placentation hémochoriale qui a eu lieu chez les Hyaenidae. Ensuite, nous avons cherché des gènes syncytine dans le genre non-mammifère Mabuya, des lézards vivipares présentant un type rare de placenta très complexe et proche de celui des mammifères. Nous avons identifié une env qui a été capturée et conservée dans ce genre depuis sa radiation, il y a 25 millions d'années. Ce gène, que nous avons appelé syncytin-Mab1, est capable d'induire la fusion cellule-cellule et est exprimé dans une couche de cellules fusionnées à l'interface materno-fœtale du placenta, deux propriétés canoniques de syncytine. Nous avons aussi identifié le récepteur de syncytin-Mab1, MPZL1, et avons montré que leur interaction induit son activation et sa phosphorylation. L'activation de MPZL1 a été liée à la migration et à l'invasion cellulaire, indiquant que cette interaction env-récepteur pourrait jouer un rôle dans l'invasion placentaire du tissu maternel observée chez les Mabuya. Pour conclure, la caractérisation de ces deux nouvelles env indique que les gènes de type syncytine ont pu jouer un rôle à la fois dans l'émergence du placenta de Mabuya et dans la structure atypique du placenta des hyènes, supportant la notion que la capture d'env est une force évolutive majeure. / Syncytins are captured retroviral envelope genes (env) that are essential for the establishment of placental structures in mammals. The syncytins present in different mammalian families are highly diverse, resulting from distinct capture events, and it has been suggested that this might play a role in making the placenta the most diverse structure in mammals. Here we used two different approaches to investigate the links between env capture and emergence and diversity of placental structures. First, we investigated placentation in Hyaenidae, the only carnivorans that present a highly invasive hemochorial placenta, as is also found in humans. Hyenas express the previously identified syncytin-Car1 gene, as do all carnivorans, but we identified a new hyena-specific captured env that we named Hyena-Env2. This new gene is present at the same locus in all hyenas, having been captured during the radiation of this family. It is non-fusiogenic but still conserved over at least 10 million years of evolution and expressed at the materno-fetal interface in the hyena placenta, making it a candidate gene for explaining the endotheliochorial to hemochorial placental transition that occurred in Hyeanidae. Second, we searched for syncytin-like genes in the non-mammalian Mabuya lizards, which are viviparous and present a rare type of highly complex placenta that is very reminiscent of mammalian placentas. We identified an env gene that was captured and conserved in this genus since its radiation 25 million years ago. This gene, that we named syncytin-Mab1, is able to mediate cell-cell fusion in vitro and is expressed in a fused cell layer at the materno-fetal interface of the placenta in vivo, characteristic features of canonical mammalian syncytin genes. We also identified the cellular gene MPZL1 as the cognate receptor of syncytin-Mab1 and showed that their interaction induces activation and phosphorylation of the former. MPZL1 activation has been linked with cell migration and invasion, indicating that this env-receptor interaction could play a role in the placental invasion of maternal tissues observed in Mabuya. In conclusion, the characterization of these two novel env genes indicates that syncytin-like env might have played a role both in the emergence of the Mabuya placenta and the atypical placental structure of hyenas, reinforcing the notion that env capture is a major driving force in evolution.

: L’économie de la filière équine repose aujourd’hui sur la production de chevaux athlètes performants sur le long terme. Le métabolisme de la jument gestante peut programmer le développement du poulain, sa santé à long terme et donc ses performances sportives à l’âge adulte. De nombreuses pratiques d’élevage peuvent modifier le métabolisme maternel, telles que la nutrition durant la gestation, la surnutrition durant la vie de la jument (surpoids et obésité) et le nombre de poulains produits par la jument (parité). L’objectif de ce travail était d’étudier les effets du métabolisme maternel durant la gestation sur la fonction et la structure placentaire à terme, la croissance osseuse, le métabolisme énergétique, l’inflammation systémique et le statut ostéoarticulaire des poulains en croissance. Un premier modèle de perturbation nutritionnelle en fin de gestation a été développé en comparant des juments ayant ingéré uniquement des fourrages au cours de la gestation ou bien des fourrages et des concentrés à partir de la mi-gestation. Ce modèle a permis de montrer que la supplémentation en concentrés altérait le métabolisme glucidique maternel, la fonction placentaire ainsi que le statut ostéoarticulaire et la réponse métabolique à un challenge de surnutrition chez le poulain. D’autre part, une perte d’état trop importante associée à une qualité/quantité de foin insuffisante entrainait un retard de maturité des fonctions de métabolisme énergétique et de reproduction mâle chez les poulains. Un deuxième modèle a ensuite été développé pour étudier l’effet de la primiparité. Cette étude a confirmé que la croissance fœtale des poulains issus des juments primipares était réduite et que ces poulains demeuraient plus petits avec un métabolisme glucidique et une maturation testiculaire retardés par rapport aux poulains issus de juments multipares. Le troisième modèle développé s’est intéressé à l’effet de l’obésité maternelle dès la conception. En effet, la prévalence de surpoids et d’obésité est de plus en plus importante au sein de la filière équine. Ce dernier modèle a permis de montrer que l’obésité maternelle associée à une résistance à l’insuline et une inflammation systémique augmentées entrainait une augmentation de la résistance à l’insuline, de l’inflammation systémique et du développement de lésions d’ostéochondrose chez les poulains. L'ensemble de ces résultats met en avant la relation entre la résistance à l’insuline maternelle, l’inflammation maternelle et le développement de lésions d’ostéochondrose chez les poulains durant la croissance, mais également entre sous-nutrition utérine et retard de maturité. Ces observations vont permettre de développer de nouvelles recommandations nutritionnelles pour les poulinières. / The economy of the equine industry is based on the production of high performance athlete horses. The metabolism of the pregnant mare can program the development of the foal, its long-term health and therefore its athletic performance at adulthood. Many breeding practices can modify maternal metabolism, such as nutrition during pregnancy, overnutrition during the mare's productive life (overweight and obesity) and the number of foals produced by the mare (parity). The aim of this work was to study the effects of maternal metabolism during pregnancy on placental function and structure, as well as bone growth, energy metabolism, systemic inflammation and osteoarticular status in growing foals. In a first approach, mares fed with forage only during gestation were compared to mares fed forage and concentrates from mid-gestation. Supplementation with concentrates altered maternal carbohydrate metabolism and placental function. In weaned foals, the osteoarticular status and the metabolic response to an overnutrition were also affected by the use of concentrates in maternal nutrition. Conversely, mares fed forage only lost body condition, which led to a delay in the post-natal maturation in terms of energy metabolism and testicular function in foals. In a second approach, the effect of primiparity was studied. Foals born to primiparous mares were growth restricted at birth and had a long-term maturational delay in bone growth, carbohydrate metabolism and testicular function. Finally, given the current high prevalence of overweight and obesity in the equine species, the effects of maternal obesity were studied. Maternal obesity associated with increased maternal insulin resistance and systemic inflammation resulted in increased insulin resistance, systemic inflammation, and increased incidence of osteochondrosis in foals. Altogether, these results highlight the relationship between maternal insulin resistance, maternal inflammation and the development of osteochondrosis lesions in foals during growth, but also between in utero undernutrition and maturation delay. These observations will contribute to adjust nutritional recommendations to broodmares.

La pollution atmosphérique est un problème de santé publique majeur responsable en 2012 de 3,7 millions de décès prématurés dans le monde. La pollution de l’air, au même titre que les autres facteurs environnementaux (e.g., nutrition, stress) auxquels sont exposées les femmes enceintes, pourrait avoir un impact sur le développement foetal. L’objectif de ce travail était d’étudier les effets d’une exposition gestationnelle aux gaz d’échappement de moteur diesel, source majeure de la pollution atmosphérique, sur le développement foetoplacentaire en première (F1) et deuxième (F2) génération. Des lapines gestantes (F0) ont été exposées 2h/jour du 3e au 27e jour post-conception (jpc) à des niveaux de gaz d’échappement de moteur diesel mimant un pic de pollution aux particules fines de grandes villes européennes. L’exposition gestationnelle des lapines F0 entrainait des signes d’hypotrophie foetale (réduction de la longueur de la tête, de l’efficacité placentaire et hypoinsulinémie des foetus F1 à 28jpc) associés à un défaut de vascularisation placentaire et des dérégulations fonctionnelles du placenta identifiées par une analyse d’enrichissement de réseaux de gènes (GSEA) sur des données de transcriptomique (microarray dédié). Le transfert transplacentaire des nanoparticules inhalées a été démontré par microscopie électronique à transmission. En F2, la concentration plasmatique en triglycérides était réduite et la concentration plasmatique en cholestérol augmentée chez les foetus issus des lapines F1 exposées in utero. La biométrie foeto-placentaire et la structure du placenta n’ont pas été affectées. Les analyses GSEA du transcriptome placentaire F2 ont révélé des perturbations de réseaux de gènes impliqués dans la formation du protéasome et desexosomes et dans l’inflammation. Les modulations de profils en acides gras au niveau placentaire et fœtal suggèrent la mise en place de mécanismes adaptatifs anti-inflammatoires chez les F2. En conclusion, l’exposition gestationnelle chronique à des gaz d’échappement de moteur diesel induit des perturbations foeto-placentaires sur plusieurs générations. Ces données ont mis en évidence le risque lié aux nanoparticules émises par les moteurs diesel et qui ne sont soumises actuellement à aucune réglementation. / Atmospheric pollution is a major threat for human health, causing 3.7 million premature deaths worldwide in 2012. Air pollution, as well as other environmental factors (e.g., nutrition, stress) faced by pregnant women may affect fetal development. The objective of this work was to study the impact of gestational exposure to diesel engine exhaust (DE), a major source of air pollution, on the fetoplacental development in the first (F1) and second (F2) generation. Pregnant rabbits (F0) were exposed 2h/day from 3 to 27 days post-conception (dpc) at DE levels mimicking a pollution peak in major European cities. Gestational exposure of F0 females induced signs of fetal hypotrophia (reduced head length, placental efficiency and hypoinsulinemia in fetuses at 28 dpc) associated with placental vasculature failure and placental functional deregulations identified by gene set enrichment analysis (GSEA) on transcriptomic data (dedicated microarray). Transplacental transfer of inhaled nanoparticles was demonstrated by transmission electronic microscopy. In F2 generation, plasma triglyceride concentrations were reduced and plasma cholesterol concentration increased in fetuses of in utero exposed F1 rabbits. Placental biometry and structure were not affected. GSEA analysis of F2 placental transcriptome revealed disturbances in gene networks involved in formation of proteaome and exosome, in inflammation. Modulations of placental and fetal fatty acid profiles suggest anti-inflammatory adaptive mechanisms in F2 fetuses. In conclusion, chronic gestational exposure to diesel engine exhaust affects fetoplacental development over several generations. These data highlight health hazards caused by DE nanoparticles and currently not subject to regulation.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and toxikology Student: Zuzana Matiašková Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Title of diploma thesis: Role of selected ABC and SLC transporters in transmembrane permeability of maraviroc: effect on transport in placenta Antiretroviral drug maraviroc is an inhibitor of CCR5-trophic HIV virus and belongs to the group of entry inhibitors. Nowadays, maraviroc is administered as part of combination antiretroviral therapy (cART) primarily in adults, children over the age of two and pregnant women to reduce the risk of transmission of HIV to the fetus. The knowledge of interactions of maraviroc with drug transporters in placenta is crucial for optimizing the therapy during pregnancy, both in terms of efficacy and potential adverse effects. Maraviroc is known substrate of ABCB1 transporter, which plays a protective role to the fetus by its efflux activity in the apical membrane of trophoblast. However, the results of recent study employing dually perfused human placenta suggest involvement of other transport mechanisms in the maraviroc transplacental pharmaocokinetics, especially those operating in the opposite direction to ABCB1. The aim of this study was to evaluate in vitro studies whether, besides ABCB1,...