Ketoprofen is a potent non-steroidal anti-inflammatory drug which is used for the treatment of rheumatoid arthritis. The oral administration of ketoprofen can cause gastric irritation and adverse renal effects. Transdermal delivery of the drug can bypass gastrointestinal disturbances and provide relatively consistent drug concentrations at the site of administration. The release of ketoprofen from proprietary gel products from three different countries was evaluated by comparing the in vitro release profiles. Twenty extemporaneously prepared ketoprofen gel formulations using Carbopol® polymers were manufactured. The effect of polymer, drug concentration, pH and solvent systems on the in vitro release of ketoprofen from these formulations were investigated. The gels were evaluated for drug content and pH. The release of the drug from all the formulations obeyed the Higuchi principle. Two static FDA approved diffusion cells, namely the modified Franz diffusion cell and the European Pharmacopoeia diffusion cell, were compared by measuring the in vitro release rate of ketoprofen from all the gel formulations through a synthetic silicone membrane. High-performance liquid chromatography and ultraviolet spectrophotometric analytical techniques were both used for the analysis of ketoprofen. The validated methods were employed for the determination of ketoprofen in the sample solutions taken from the receptor fluid. Two of the three proprietary products registered under the same manufacturing license exhibited similar results whereas the third product differed significantly. Among the variables investigated, the vehicle pH and solvent composition were found have the most significant effect on the in vitro release of ketoprofen from Carbopol® polymers. The different grades of Carbopol® polymers showed statistically significantly different release kinetics with respect to lag time. When evaluating the proprietary products, both the modified Franz diffusion cell and the European Pharmacopoeia diffusion cell were deemed adequate although higher profiles were generally obtained from the European Pharmacopoeia diffusion cells. Smoother diffusion profiles were obtained from samples analysed by high-performance liquid chromatography than by ultraviolet spectrophotometry in both diffusion cells. Sample solutions taken from Franz diffusion cells and analysed by ultraviolet spectrophotometry also produced smooth diffusion profiles. Erratic and higher diffusion profiles were observed with samples taken from the European Pharmacopoeia diffusion cell and analysed by ultraviolet spectrophotometry. The choice of diffusion cells and analytical procedure in product development must be weighed against the relatively poor reproducibility as observed with the European Pharmacopoeia diffusion cell.

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Martina Kobrlová Supervisor: prof. PharmDr. Petr Pávek, Ph.D. Title of diploma thesis: Pharmacogenetics in rheumatoid arthritis Based on scientific progress in the research of human genome and the discovery of polymorphisms, which are involved in the interindividual differences in human population, there is also a growing interest in pharmacogenetics. It is a field combining pharmacology and genetics with the aim of identifying specific features that could explain the different responses of patients to treatment by clinically used drugs. Applying this knowledge could contribute to a simpler choice of medication for a particular patient and it could reduce the risk of side effects or poor response. In this diploma thesis I dealt with the latest scientific knowledge on pharmacogenetics in rheumatology, in particular the rheumatoid arthritis. From available studies, reviews, and meta-analyzes that have been published, I summarized current data on the relationship between polymorphisms and disease modifying drugs (DMARDs) used for the treatment of this disease. The largest amount of data was found on the most commonly used methotrexate. Further, the work examines the leflunomide and other...

Reumatoid artrit är en progressiv inflammatorisk sjukdom som bland annat orsakar smärta och fatigue i kroppen där hela personen och dess liv påverkas. Brist på kunskap om hur det är att leva med reumatoid artrit kan leda till att personer inte får den vård de behöver. Syftet med litteraturstudien var att belysa erfarenheter av att leva med reumatoid artrit. Studien genomfördes som en litteraturstudie. Databassökningar gjordes systematiskt med sökord som var relevanta utifrån syftet. Litteraturstudien baserades på åtta kvalitativa artiklar som kvalitetsgranskades och analyserades, vilka resulterade i två teman, begränsningar och strategier. Erfarenheterna av att leva med reumatoid artrit var att symtomen begränsade personen i det dagliga livet. För att hantera begränsningarna och ta sig vidare i livet krävdes planering och acceptans. Genom att fokusera på det som var bra insåg personerna att de är så mycket mer än sjukdomen. Personcentrerad vård är en av sjuksköterskans kärnkompetenser som fokuserar på att se hela människan utifrån respekt och lyhördhet vilket leder till individualiserad vård där personer känner sig sedda och hörda. / Rheumatoid arthritis is a progressive inflammatory disease that causes pain and fatigue. The whole individual and their life is affected by living with rheumatoid arthritis. Lack of knowledge about living with rheumatoid arthritis can lead to that people don´t get the care they need. The purpose of the literature study was to highlight experiences of living with rheumatoid arthritis. The study was conducted as a literature study. Database searches were made systematically using keywords relevant to the purpose. The literature study was based on eight qualitative articles which were analyzed and graded for quality. The themes of the result were limitations and strategies. The experience of living with rheumatoid arthritis was that the symptoms limited the person and their daily life. To manage the constraints and move on in life, planning and acceptance were required. By focusing on what was good, the people realized that they were so much more than the disease. Person-centered care is one of the nurse's core competencies, focusing on seeing the whole human being based on respect and responsiveness, which leads to individualized care where people feel seen and heard.

OBJETIVOS: Avaliar o perfil e o número absoluto e percentual de células NK verdadeiras (CD56+CD16+CD3-) e de células NK e NKT (CD56+) no sangue periférico de uma coorte de pacientes com artrite reumatóide (AR) antes e durante o tratamento com rituximabe (RTX). MÉTODOS: Foram analisados dez pacientes do grupo controle (doadores de sangue) e dez pacientes com AR que receberam duas infusões de RTX 1g separadas por intervalo de 14 dias. As análises imunofenotípicas para avaliação do perfil e quantificação de células NK foram realizadas pré e após a infusão ou até a recaída clínica. Pacientes respondedores e não respondedores foram classificados de acordo com os critérios do Colégio Americano de Reumatologia (ACR) em 6 meses. RESULTADOS: A quantidade de células NK verdadeiras não demonstrou variação significativa pré e após o tratamento com RTX. Contudo, houve aumento percentual de células CD56+ entre o primeiro e o segundo mês após a infusão com RTX. Além disso, os pacientes respondedores apresentaram uma tendência de aumento do número absoluto de células NK verdadeiras após dois meses de tratamento. Já em relação ao grupo controle, observou-se um aumento significativo do número de células NK basais nos pacientes com AR (p<0,05). CONCLUSÕES: Foi identificada uma tendência de aumento nos valores absolutos de células NK verdadeiras entre os pacientes respondedores no segundo mês após a infusão com RTX. Não foi identificada uma variação significativa no perfil e quantidade de células NK nos pacientes com AR pré e após o tratamento com RTX. Contudo, foi observado que os pacientes com AR possuem uma quantidade maior de células NK do que os controles, sugerindo um possível envolvimento destas células na AR. / OBJECTIVES: To assess the profile as well as the absolute number and percentage of true NK cells (CD56+CD16+CD3-), and NK and NKT cells (CD56+) in the peripheral blood of a cohort of patients with rheumatoid arthritis (RA) before and during rituximab (RTX) therapy. METHODS: Ten control patients (blood donors) and ten patients with RA were assessed. The latter group received two intravenous infusions of 1g RTX, separated by a 14 day interval. Immunophenotypic analyses of NK cells were conducted before and after infusion, or until clinical relapse. After six months, respondents and nonrespondents were reassessed according to American Rheumatology Criteria (ARC). RESULTS: The number of true NK cells did not significantly change after treatment with RTX. However, an increase in the percentage of CD56+ cells was observed between the first and second month after RTX infusion. Respondents also displayed a tendency toward an increased number of true NK cells after two months of treatment. At baseline, the number of NK cells was also found to be significantly higher in patients with RA than in control individuals (p<0.05). CONCLUSIONS: Respondents displayed a tendency toward an increase in the absolute number of true NK cells in the second month after RTX infusion. No significant changes in the profile and frequency of NK cells were found between preand post-RTX treatment assessments of patients with RA. However, it was found that patients with RA have a higher number of NK cells than control partcipants, suggesting a possible role of these cells in RA.

Submitted by Nadir Basilio (nadirsb@uninove.br) on 2018-06-21T15:40:31Z No. of bitstreams: 1 Ana Carolina Araruna Alves.pdf: 1155545 bytes, checksum: 5e9fda24285778105d5ac0d8276f9036 (MD5) / Made available in DSpace on 2018-06-21T15:40:31Z (GMT). No. of bitstreams: 1 Ana Carolina Araruna Alves.pdf: 1155545 bytes, checksum: 5e9fda24285778105d5ac0d8276f9036 (MD5) Previous issue date: 2016-03-29 / Rheumatoid Arthritis (RA) treatment in the last decades has evolved medications with cytokine inhibitors, which though they are today considered the gold standard in RA treatment, still do not guarantee complete remission from the disease or from hyperalgesia, and it is know that the Photobiostimulation (PBM) is also capable of acting on the expression of pro-inflammatory cytokines, being in this way an alternative treatment for RA. This research had as its objective the analysis and comparison of the effects of PBM in the acute phase of experimental RA in terms of nociception, of the number of inflammatory cells (neutrophils, macrophages, lymphocytes), and of the protein expression of inflammatory mediators (TNF-α, IL-6, IL-10), in the lavage and joint cartilage of rats. 49 male Wistar rats were used, with an approximate age of 90 days and a body weight varying from 250 to 300g. The animals were randomly distributed in 3 groups, with 21 animals in each experimental group (Rheumatoid arthritis group – RA) and the group treated with PBM (RA-LLLT), and with 7 animals in the control group. Each group was arrranged in three distinct experimental periods according to the time of euthanasia (6 hours, 24 hours, and 48 hours). For the reproduction of the lesion the animals were submitted to two inductions: the first consisted in a subcutaneous infiltration in the dorsal of each animal (in the RA and RA-LLLT groups) using 500 µg of mBSA diluted in a solution of 100 µL of complete Adjuvant de Freund (CFA), and 100 µL of PBS, with this procedure being repeated weekly for 21 days. On the 28th day, an intrajoint induction was carried out with 10 µg of mBSA diluted in 10µL of PBS, in the joint space of the right hind paw of each animal. After the joint induction, the treatment on the RA-LLLT group began, using a DMC® brand Photon Laser III, with a wave length of λ 808 nm, active medium of Gallium Arsenide, and Aluminium (AsGaAI), with a potency of 50 mW (potency density of 1,78 W/cm2) and area of the beam 0,028cm2. The final dose was of 2J, the density of energy of 71,2 J/cm2, and time of 40s by point. At the end of each test period (6 hours, 24 hours, and 48 hours) the animals were submitted to an evaluation of their nociception, followed by the extraction of joint lavage and joint cartilage, which were sent for analysis of the total count and differential cells, and for protein expression of the inflammatory mediators described. The results demonstrate biomodulation in the expression of neutrophils and macrophages amongst the control group and RA, as well as in the cytokines IL-6 and IL-10, contributing daily to the attenuation of hyperalgesia by the PBM. / O tratamento para Artrite Reumatoide (AR) com inibidores de citocinas tem sido considerado padrão-ouro nas últimas décadas embora ainda não garantam a remissão completa da doença ou da hiperalgesia, sabe-se que a Laserterapia de Baixa Intensidade (LBI) também é capaz de atuar na expressão das citocinas pró-inflamatórias, podendo ser assim, uma alternativa de tratamento para a AR. Este trabalho teve como objetivo analisar os efeitos da LBI na fase aguda de AR experimental sobre a nocicepção, sobre o número de células inflamatórias (neutrófilos, macrófagos, e linfócitos), e sobre a expressão proteica de mediadores inflamatórios (TNF-α, IL-6, IL-10) no lavado e cartilagem articular de ratos. Foram utilizados 49 ratos Wistar, machos, com idade aproximada de 90 dias e peso corporal variando de 250 a 300 g. Os animais foram distribuídos aleatoriamente em 3 grupos, com 21 animais em cada grupo experimental (grupo Artrite Reumatoide - RA e o grupo tratado com laser AR-LBI), e 7 animais no grupo controle. Cada grupo foi composto por 3 tempos experimentais distintos de acordo com o tempo da eutanásia (6 horas, 24 horas e 48 horas). Para a reprodução da lesão os animais foram submetidos a duas induções: a primeira consistiu em uma infiltração subcutânea no dorso de cada animal (nos grupos AR e AR-LBI) utilizando-se 500 µg de mBSA diluído em uma solução de 100 µL de Adjuvante completo de Freund (CFA), e 100µL de PBS, esse procedimento foi repetido semanalmente por 21 dias. No 28º dia, foi realizada uma indução intra-articular com 10 µg de mBSA diluído em 10µL PBS, no espaço articular da pata direita traseira de cada animal. Após a indução articular, deu-se início ao tratamento no grupo AR-LBI, por meio do laser da marca DMC® modelo Photon Laser III, com comprimento de onda de λ 808 nm, meio ativo de Arsenieto de Gálio e Alumínio (AsGaAI), com potência de 50 mW (densidade de potência de 1,78 W/cm2), área do feixe de 0,028cm2 e aplicação sob a forma de dois pontos pelo método transcutâneo nos compartimentos medial e lateral da articulação, dose final foi de 2J, densidade de energia de 71,2 J/cm2, e tempo 40s, por ponto. Ao final de cada período experimental (6 horas, 24 horas e 48 horas) os animais foram submetidos à avaliação da nocicepção, seguido de extração do lavado articular e da cartilagem articular, que foram encaminhados às análises de contagem total e diferencial de células, e da expressão proteica dos mediadores inflamatórios descritos. Os resultados demonstram biomodulação da LBI principalmente na expressão de neutrófilos e macrófagos entre os grupos controle e AR, bem como nas citocinas IL-6 e IL-10, contribuindo diretamente para a atenuação da hiperalgesia.

Nous avons développé des nanoparticules d’une prodrogue de glucocorticoïde, la dexaméthasone palmitate (DXP) à visée thérapeutique dans le traitement de la polyarthrite rhumatoïde (PR). Cette maladie auto-immune est caractérisée par une inflammation articulaire, une érosion osseuse et cartilagineuse et une dérégulation du système immunitaire. Parmi les traitements indiqués dans la PR, l'utilisation des glucocorticoïdes est limitée par leurs effets secondaires importants induits par leur pharmacocinétique défavorable. Afin de traiter la PR par voie intraveineuse, la formulation de nanoparticules PEGylées semble indispensable afin d’échapper au phénomène d’opsonisation et d’espérer obtenir une accumulation spécifique au niveau des articulations inflammées. Pour cela, nous avons développé des nanoparticules de DXP (DXP-NP) stabilisées par le DSPE-PEG2000.Les caractéristiques physico-chimiques des nanoparticules obtenues ont été évaluées ainsi que leur stabilité au cours du temps. La structure interne des nanoparticules définie comme amorphe ainsi que leur très fort taux de charge prouvent ainsi l’impact du DSPE-PEG2000 dans l’organisation moléculaire des DXP-NP. In vivo, l’étude de la pharmacocinétique et de la biodistribution des DXP-NP suite à leur administration intraveineuse a démontré une circulation prolongée du système. Dans un modèle murin de polyarthrite rhumatoïde, les DXP-NP ont démontré leur accumulation spécifique dans les articulations inflammées en corrélation avec une supériorité thérapeutique significative en comparaison avec la molécule libre hydrosoluble. Des études histologiques ainsi que l’évaluation du traitement sur l’apparition d’effets indésirables complètent l’étude in vivo. / We developed nanoparticles of a glucocorticoid prodrug, dexamethasone palmitate (DXP) for the treatment of rheumatoid arthritis (RA). Joint inflammation, bone and cartilage erosion and dysregulation of the immune system characterize this autoimmune disease. Among the treatments indicated in RA, the use of glucocorticoids is hampered by their side effects induced by their unfavorable pharmacokinetics. To treat RA intravenously, the formulation of PEGylated nanoparticles seems essential in order to escape from opsonization and to obtain a specific accumulation in the joints inflamed. Therefore, we developed DXP nanoparticles (DXP-NPs) stabilized by the DSPE-PEG2000.The physicochemical characteristics of the nanoparticles obtained were evaluated as well as their stability over time. The amorphous internal structure of the nanoparticles and their very high drug loading thus prove the impact of DSPE-PEG2000 in the molecular organization of DXP-NPs. In vivo, the study of the pharmacokinetics and biodistribution of DXP-NPs following intravenous administration demonstrated prolonged circulation of the system. In a mouse model of rheumatoid arthritis, DXP-NPs their demonstrated specific accumulation in inflamed joints in correlation with significant therapeutic superiority in comparison with the water-soluble free molecule. Histological studies as well as adverse events evaluation supplemented the in vivo study.

Ample studies have shown that perturbation of metabolic phenotype is correlated with gene mutation and pathogenesis of colorectal cancer (CRC) and rheumatoid arthritis (RA). Mass spectrometry (MS)-based metabolomics as a powerful and stable approach is widely applied to bridge the gap from genotype/metabolites to phenotype. In CRC suffers, KRAS mutation accounts for 35%-45%. In previous study, SLC25A22 that encodes the mitochondrial glutamate transporter was found to be overexpressed in CRC tumor and thus to be essential for the proliferation of CRC cells harboring KRAS mutations. However, the role of SLC25A22 on metabolic regulation in KRAS-mutant CRC cells has not been comprehensively characterized. We performed non-targeted metabolomics, targeted metabolomics and isotope kinetic analysis of KRAS-mutant DLD1 cells with or without SLC25A22 knockdown using ultra-high performance liquid chromatography (UHPLC) coupled to Orbitrap MS and tandem MS (MS/MS). In global metabolomics analysis, 35 differentially regulated metabolites were identified, which were primarily involved in alanine, aspartate and glutamate metabolism, urea cycle and polyamine metabolism. Then targeted metabolomics analysis on intracellular metabolites, including tricarboxylic acid (TCA) cycle intermediates, amino acids and polyamines, was established by using LC-MS/MS coupled with an Amide BEH column. Targeted metabolomics analysis revealed that most TCA cycle intermediates, aspartate (Asp)-derived asparagine, alanine and ornithine (Orn)-derived polyamines were strongly down-regulated in SLC25A22 knockdown cells. Moreover, the targeted kinetic isotope analysis using [U-13C5]-glutamine as isotope tracer showed that most of the 13C-labeled TCA cycle intermediates were down-regulated in SLC25A22-silencing cells. Orn-derived polyamines were significantly decreased in SLC25A22 knockdown cells and culture medium. Meanwhile, accumulation of Asp in knockdown of GOT1 cells indicated that oxaloacetate (OAA) was majorly converted from Asp through GOT1. Exogenous addition of polyamines could significantly promote cell proliferation in DLD1 cells, highlighting their potential role as oncogenic metabolites that function downstream of SLC25A22-mediated glutamine metabolism. SLC25A22 acts as an essential metabolic regulator during CRC progression as promotes the synthesis of TCA cycle intermediates, Asp-derived amino acids and polyamines in KRAS-mutant CRC cells. Moreover, OAA and polyamine could promote KRAS-mutant CRC cell growth and survival. Rheumatoid arthritis (RA) is a chronic, inflammatory and symmetric autoimmune disease and a major cause of disability. However, there is insufficient pathological evidence in term of metabolic signatures of rheumatoid arthritis, especially the metabolic perturbation associated with gut microbiota (GM). Based on consistent criteria without special diet and therapeutic intervention to GM, we enrolled 50 RA patients and 50 healthy controls. On basis of the platform of UHPLC-MS and GC-MS, were performed for the non-targeted metabolomics to investigate alterations of endogenous metabolites in response to RA inflammation and interaction with GM. 32 and 34 significantly changed metabolites were identified in urine and serum of patients with RA, respectively. The altered metabolites were identified by HMDB, METLIN database or authentic standards, and mostly metabolites were attributed into tryptophan and phenylalanine metabolism, valine, leucine and isoleucine biosynthesis, aminoacyl-tRNA biosynthesis and citrate cycle. To obtain alterations of more components in tryptophan and phenylalanine metabolism, we developed and validated a targeted metabolomics method of 19 metabolites by using LC-QqQ MS. Combining the results of targeted metabolomics with global metabolomics, significantly up-regulated kynurenine (KYN), anthranilic acid (AA) and 5-hydroxylindoleacetic acid (HIAA) simultaneously in urine and serum was found to implicate the activation of tryptophan metabolism under the condition of RA, which acted pro-inflammatory roles in inflammation and was closely correlated with GM. IDO/TDO functioned as a pro-inflammation mediator was overexpressed in RA patients. Urinary kynurenic acid and serum serotonin that have impacts on anti-inflammation in immune system were down-regulated in RA patients. The levels of phenylacetic acid and phenyllactic acid serving as a pro-inflammatory and an anti-inflammatory agent, respectively, increased in serum of patients with RA. Moreover, certain essential amino acids (EAAs), and mostly conditional EAAs were decreased in RA patients, which have been reported to inhibit cell proliferation of immune cells. In particular, deficiency of branched chain amino acids (BCAAs, valine and isoleucine) was observed in serum of patients with RA, which may lead to muscle loss and cartilage damage. The specificity of all altered metabolites resulted from RA was considerably contributed through the GM-derived metabolites. The findings revealed that GM-modulated RA inflammation was mainly resulted from tryptophan and phenylalanine metabolism, and amino acid biosynthesis, which may provide more information for better understanding the RA mechanism.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22144号 / 医博第4535号 / 新制||医||1039(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 松田 秀一, 教授 生田 宏一, 教授 杉田 昌彦 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

付記する学位プログラム名: 充実した健康長寿社会を築く総合医療開発リーダー育成プログラム / 京都大学 / 新制・課程博士 / 博士(人間健康科学) / 甲第23127号 / 人健博第89号 / 新制||人健||6(附属図書館) / 京都大学大学院医学研究科人間健康科学系専攻 / (主査)教授 杉本 直三, 教授 黒木 裕士, 教授 松田 秀一 / 学位規則第4条第1項該当 / Doctor of Human Health Sciences / Kyoto University / DFAM

La polyarthrite rhumatoïde (PR) est le rhumatisme chronique le plus fréquent caractérisé par une augmentation du risque de mortalité cardiovasculaire (CV). Ce sur-risque est la conséquence d’une athérogenèse accélérée, mais aussi d’atteintes microvasculaires, secondaires à une dysfonction endothéliale (DE). La réversibilité de la DE en fait une cible thérapeutique pertinente pour réduire la surmortalité en cas de PR. Ces dernières années, la physiopathologie de la DE au niveau des artères de conductance a été comprise grâce aux modèles animaux d’arthrite, principalement par le modèle d’arthrite induite à l’adjuvant (AIA). Toutefois, les effets du méthotrexate (MTX), traitement de première intention de la PR, sur la DE sont toujours inconnus. De même, les études sur la DE des vaisseaux coronariens et cérébraux font défaut.L’objectif de ce travail de thèse a été d’étudier les effets du MTX sur la DE aortique et de comprendre les mécanismes impliqués, mais aussi d’explorer et caractériser la DE au niveau des artères cérébrales moyennes et des artères coronaires dans le modèle AIA chez le rat.Dans une première étude, nos résultats ont montré qu’un traitement de 21 jours par MTX (1 mg/kg/semaine) n’améliore pas la DE aortique en dépit d’effets positifs surla sévérité clinique et l’inflammation systémique.Dans une seconde étude, nos données révèlent que le modèle AIA est associé à une DE au niveau des artères cérébrales moyennes, avec un rôle déterminant de la suractivité de l’arginase, comme en témoigne la capacité d’un traitement par nor-NOHA (40 mg/kg/jour), un inhibiteur d’arginase, à complétement inverser la DE cérébrovasculaire.Dans une troisième étude, nous avons démontré que le modèle AIA est caractérisé par une DE coronarienne positivement corrélée avec les taux plasmatiques d’endothéline-1 et d’angiotensine II, par une inflammation et une hypertrophie cardiaque, et une augmentation de la susceptibilité à l’ischémie myocardique.En conclusion, nos données montrent que les effets positifs du MTX sur le risque CV ne sont pas dus à une amélioration de la DE aortique, soulignant la nécessité de trouver des traitements ciblant spécifiquement le système CV en tant que traitements adjuvants des médicaments antirhumatismaux actuels. De plus, nos données montrent que le modèle AIA est un bon modèle pour mimer les atteintes cardiaques et cérébrovasculaires et pour étudier l’impact des nouveaux traitements visant à réduire le risque CV dans la PR. / Rheumatoid arthritis (RA) is the more frequent chronic rheumatism characterized by an increased risk of cardiovascular (CV) mortality. This over-risk is the consequence of accelerated atherosclerosis, but also of microvascular damages, secondary to an endothelial dysfunction (ED). The reversibility of the ED makes of it a relevant therapeutic target to reducing excessive mortality in RA. In recent years, pathophysiology of ED in conductance arteries has been understood thank to animal models of arthritis, mainly the adjuvant-induced arthritis (AIA) model in rats. However, the effects of methotrexate (MTX), first-line treatment in RA, on ED are still unknown. Likewise, studies on ED of the coronary and cerebral vasculature are lacking.The aim of this thesis work was to study the effect of MTX on aortic ED and to dissect the mechanisms involved, but also to explore and characterize ED on MCA and coronary arteries in the widely-used model of adjuvant-induced arthritis (AIA) in rats.In a first study, our results showed that a 21-days treatment with MTX (1 mg/kg/week) did not improveaortic ED despite positive effects on clinical severity and systemic inflammation.In a second study, our data reported that the AIA model was associated with ED in middle cerebral arteries, involving a seminal role of arginase upregulation, as attested by the capacity of a treatment with nor-NOHA (40 mg/kg/day), an arginase inhibitor, to fully reverse cerebrovascular ED.In a third study, we demonstrated that the AIA model was characterized by a coronary ED positively correlated to plasma levels of endothelin-1 and angiotensin II, by a cardiac inflammation and hypertrophy, and an increased susceptibility to myocardial ischemia.In conclusion, our data indicate that positive effects on CV risk of MTX do not involve improving aortic ED, underlying the necessity to find accurate therapies targeting specifically the CV system as adjuvant treatments to the current anti-rheumatic drugs. Moreover, our data showed that the AIA model is relevant for mimicking cardiac and cerebrovascular impairments, and to study the impact of new treatments for reducing CV risk in RA.