Le travail décrit dans cette thèse a commencé avec la découverte d’effets anti-inflammatoires chez certaines souches de L. delbrueckii. Il avait été montré que l’effet anti-inflammatoire est souche-dépendant, et implique l’action de protéines exposées à la surface de la bactérie. Dans le but d’identifier l’effecteur bactérien à l’origine de l’effet immuno-modulateur, 8 souches de L. delbrueckii ont été sélectionnées. Deux de ces souches sont à fort effet anti-inflammatoires, et les 6 restantes sont à effet faible ou intermédiaire. Pour l’identification des protéines potentiellement responsables pour l’effet anti-inflammatoire, des études de génomique et transcriptomique comparatives des 8 souches de L.delbrueckii ont été entreprises, ainsi qu’une étude comparative du protéome de surface bactérienne.La première partie de cette thèse décrit les résultats de finition du génome d’une des deux souches hautement anti-inflammatoires. Cette étape a révélé que la partie manquante de la séquence génomique était principalement composée de séquences répétées de type séquences d’insertions (IS), dont le nombre s’avère particulièrement élevé.La deuxième partie de la thèse décrit une étape de valorisation des données génomiques à travers une étude comparative entre souches de la ssp. lactis et souches de la ssp. bulgaricus. Cette étude révèle que les deux ssp. de L. delbrueckii évoluent en adaptation au milieu lait. Toutefois, la ssp. bulgaricus semble avoir atteint un stade d’adaptation plus avancé que celui de la ssp. lactis. L’adaptation des deux ssp. à leur environnement se fait principalement par un phénomène de perte spontanée de gènes devenus superflus.Une étude plus avancée de la structure génomique des deux ssp. révèle deux nouveaux aspects des différences de structure génomique. Tout d’abord, au sein du core génome des deux sous-espèces, les évènements d’échange génétique et recombinaison ont contribué plus à la diversité au sein de la ssp. lactis qu’à la diversité chez la ssp. bulgaricus. Ensuite, une structure inversée répétée de grande taille, rarement observée dans les génomes bactériens, s’avère caractéristique de la ssp. bulgaricus. La troisième partie de thèse décrit les trois approches comparatives menées dans le but d’identifier les protéines bactériennes à l’origine de l’effet anti-inflammatoire. Au bout de ces études, nous avons sélectionné 56 gènes candidats, dont 41 ont été clonés dans un système d’expression hétérologue. Pour l’instant, 17 clones d’expression ont été testés in vitro pour leur potentiel immuno-modulateur. Les résultats préliminaires ont permis l’identification d’une protéine à effet anti-inflammatoire. / The work described in this thesis began with the discovery of anti -inflammatory effects in certain strains of L. delbrueckii. The anti- inflammatory effect had been shown to be strain - dependent, and implicate the action of bacterial surface exposed proteins.In order to identify the bacterial effector responsible for the immunomodulatory effect, eight strains of L. delbrueckii were selected. Two of these strains have a strong anti -inflammatory effect, whereas the remaining strains have a weak to intermediary effect. To identify proteins that may be responsible for the anti- inflammatory effect, comparative genomic and transcriptomic studies of the 8 L.delbrueckii strains were conducted, as well as a comparative study of the bacterial surface proteome.The first part of this thesis describes the genome finishing of one of the highly anti- inflammatory strains. This step revealed that the missing part of the genome sequence was mainly composed of repeated sequences such as insertions sequences (IS), that were present in a particularly high number.The second part of the thesis describes the valorisation of genomic data through a comparative study between ssp. lactis strains and ssp. bulgaricus strains. This study reveals that both L. delbrueckii ssp. are evolving in adaptation to the milk environment. However, the ssp. bulgaricus appears to have reached a more advanced stage of adaptation than the ssp. lactis. The adaptation of both ssp. to their environment is primarily a phenomenon of spontaneous loss of genes that have become superfluous. A more detailed study of the genome structure of the two ssp. reveals two important differences. Firstly, within the L. delbrueckii core genome, genetic exchange and recombination contributed much more to the ssp. lactis diversity than to the ssp. bulgaricus diversity. Furthermore, a large inverted repeat structure, rarely observed in bacterial genomes, appears to be characteristic of the ssp. bulgaricus.The third part of the thesis describes the three comparative approaches used to identify bacterial proteins responsible for the anti- inflammatory effect.We selected 56 candidate genes, of which 41 were cloned in a heterologous expression system. So far, 17 expression clones were tested in vitro for their immunomodulatory potential. The preliminary results allowed the identification of one protein with anti-inflammatory effects.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22830号 / 医博第4669号 / 新制||医||1047(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 江藤 浩之, 教授 武藤 学, 教授 伊藤 貴浩 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Rheumatoid arthritis (RA) is a chronic inflammatory condition of unknown etiology for which there is no cure. It is one of the most disabling diseases and affects about 1% of the world‟s population. Recent developments in the field of molecular biology have resulted in the production of new drugs used in the treatment RA. Despite these advancements, achieving optimal disease control and prevention of disease progression is still difficult in many patients, leading to a continued search for treatment methods that will improve patient outcomes. Non-biologic forms of treatment that are still being investigated include the use of statins as an adjunct therapy due to their reported antiinflammatory effects. Some studies have shown that the use of statins in patients with RA help in reducing disease activity and swollen joint count in addition to lowering cardiovascular risk. As evidence continue to increase on the anti-inflammatory effect of statins, researchers have started investigating possible benefits that may result from the use of statins in treatment of RA, a chronic disease marked by high levels of systemic and local inflammation. This study investigated the anti-inflammatory effect of statins in rheumatoid arthritis patients with associated risks for cardiovascular disease, using simvastatin as the investigational product. Patients with moderately active RA despite being on maximum disease-modifying antirheumatic drugs (DMARDs) therapy and having associated risks for cardiovascular disease were screened for the study. Eligible patients were randomized into two groups, 1 and 2. Patients in group 1 received simvastatin treatment (20mg/day) for a period of 3 months in addition to their usual DMARDs after which they stopped simvastatin treatment and were followed up for a further 3 months off simvastatin treatment. Those in group 2 were allowed to continue on their usual DMARDs without simvastatin treatment for the first 3 months of the study after which they received 20mg/day simvastatin for a period of 3 months in addition to their usual DMARDs. The anti-inflammatory effect of simvastatin was assessed by monitoring the inflammatory variables; erythrocyte sedimentation rate (ESR) and c-reactive protein (CRP) and disease activity in the two groups at screening, at the crossover point and at end of the study. Disease activity was significantly reduced with simvastatin treatment in the two groups. The mean change in disease activity score with simvastatin treatment was 1.30 (p = 0.01) in group 1 and 1.74 (p = 0.01) in group 2. ESR was significantly reduced with simvastatin treatment in group 1 with a mean change of 19.0 (p = 0.005) and marginally reduced in group 2 with a mean change 26.0 (p = 0.09). There was no significant change in CRP with simvastatin treatment in the two groups. The mean change in CRP with simvastatin treatment was 7.0 (p = 0.24) in group 1 and 14.7 (p =0.20) in group 2. All the patients benefited from the lipid-lowering effect of simvastatin. These findings suggest that statins may have mild anti-inflammatory properties and will be good adjuvant in RA patients with associated risks for cardiovascular disease. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Pharmacology / unrestricted

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-09-27T21:13:17Z No. of bitstreams: 2 Dissertação - Patricia Antônia Vasconcelos - 2016.pdf: 2072536 bytes, checksum: 9bdd8a9eab646f3aefc3f2e50a4429e6 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-09-28T12:21:06Z (GMT) No. of bitstreams: 2 Dissertação - Patricia Antônia Vasconcelos - 2016.pdf: 2072536 bytes, checksum: 9bdd8a9eab646f3aefc3f2e50a4429e6 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-09-28T12:21:06Z (GMT). No. of bitstreams: 2 Dissertação - Patricia Antônia Vasconcelos - 2016.pdf: 2072536 bytes, checksum: 9bdd8a9eab646f3aefc3f2e50a4429e6 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-09-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Inflammation is a body protection process in response to tissue damage in order to eliminate the offending agent and promote tissue repair. It is characterized by the appearance of the classic signs such as pain, heat, redness and edema, which may occur due to loss as a result of poor resolution of the inflammatory process. The use of antiinflammatory drugs to control inflammation is widely used, but have many side effects, making necessary the search for new drugs with fewer side effects and more effective. The objective of this study was to evaluate the anti-inflammatory activity and / or antinociceptive the LQFM 147 compound using acute tests of nociception and inflammation, to characterize the mechanisms involved in such effects. For this study, male mice were used, weighing between 30 and 35g. Treatment with LQFM 147 at doses of 25, 50 and 100 mg / kg V.O. reduced the number of writhes in a dose-dependent in 16.79; 32.97 and 46.04%, respectively. In the formalin test, treatment with LQFM 147 at a dose of 50 mg / kg V.O. It reduced the time of reactivity to pain testing only the inflammatory phase 43.38%. The anti-inflammatory activity was confirmed, since the treatment with LQFM 147 at a dose of 50 mg / kg V.O. reduced formation of paw edema induced by carrageenan at all hours of the test, reduced migration of polymorphonuclear leukocytes 40.82% in pleurisy test. Treatment with LQFM 147 was able to reduce the activity of myeloperoxidase in 31.54% and TNF-α levels of 65.51%. To obtain the results the group treated with LQFM 147 was compared to the control group treated with vehicle 10 ml / kg v.o .. The data obtained demonstrate that LQFM 147 has anti-inflammatory and antinociceptive activity involving the reduction of migration of polymorphonuclear leukocytes and reduction in TNF-α levels. / Resumo: A inflamação é um processo de defesa do organismo em resposta a lesões teciduais com o objetivo de eliminar o agente agressor e promover o reparo tecidual. É caracterizada pelo aparecimento dos sinais clássicos como dor, calor, rubor e edema, podendo ocorrer a perda da função em consequência da má resolução do processo inflamatório. O uso de fármacos anti-inflamatórios para o controle da inflamação é bastante utilizado, porém possuem muitos efeitos adversos, fazendo necessária à busca por novos fármacos com menos efeitos adversos e mais eficazes. O objetivo deste trabalho foi avaliar a atividade anti-inflamatória e/ou antinociceptiva do composto LQFM 147 utilizando testes agudos de nocicepção e inflamação, visando caracterizar os mecanismos de ação envolvidos em tais efeitos. Para este estudo, foram utilizados camundongos machos, pesando entre 30 e 35 g. O tratamento com LQFM 147 nas doses de 25, 50 e 100 mg/Kg v.o. reduziram o número de contorções abdominais de maneira dose dependente em 16,79; 32,97 e 46,04%, respectivamente. No teste da formalina, o tratamento com LQFM 147 na dose de 50 mg/Kg v.o. reduziu o tempo de reatividade à dor somente na fase inflamatória do teste em 43,38%. A atividade antiinflamatória foi confirmada, uma vez que o tratamento com LQFM 147 na dose de 50 mg/Kg v.o. reduziu a formação do edema de pata induzido por carragenina em todas as horas deste teste, reduziu a migração de leucócitos polimorfonucleares em 40,82% no teste de pleurisia. O tratamento com LQFM 147 foi capaz de reduzir a atividade da enzima mieloperoxidase em 31,54% e os níveis de TNF-α em 65,51%. Para obtenção dos resultados o grupo tratado com LQFM 147 foi comparado com o grupo controle tratado com Veículo 10 mL/kg v.o.. Os dados obtidos demonstram que LQFM 147 possui atividade anti-inflamatória e antinociceptiva envolvendo a redução da migração de leucócitos polimorfonucleares e redução dos níveis de TNF-α.

In West African medicine, Entada africana Guill. & Perr. from the family of Fabaceae is used to treat inflammatory conditions in the management of fractures, wounds, and sprains in the northern region of the Republic of Benin. The aim of the present study was to isolate and elucidate phenolic compounds from a hydroalcoholic leaf extract from E. africana and to identify compounds with anti-inflammatory activity in vitro. Eleven compounds were purified from three fractions, which have shown strong to medium anti-inflammatory activity. The isolated compounds were characterized by HRESI-MS and NMR methods as gallic acid (1), ethyl gallate (2), 5,7-dihydroxychromen-4-one (3), 3′,4′,7-trihydroxyflavone (4), dihydrokaempferol-7-O-glucoside (5), catechin (6), quercetin-3-O-[β-apiosyl- (1‴→2″)-β-glucoside] (7), quercetin-3-O-glucoside (8), naringenin-7-O-glucoside (9), aromadendrin (10), and myricetin-3-O-glucoside (11). Nine of the major phenolic compounds were tested using TNF-α stimulated human keratinocytes (HaCaT) as skin inflammation model to identify molecules, which may explain the use of the plant leaves as an anti-inflammatory remedy by assessing the release of proinflammatory cytokines IL-8 and IL-6. The hydroacoholic leaf extract of E. africana exerted a medium inhibitory effect on the release of IL-8. 3′,4′,7-trihydroxyflavone, aromadendrin, dihydrokaempferol-7- O-glucoside and ethyl gallate demonstrated a strong to medium effect on the release of IL-6. For the release of IL-8, 3′,4′,7-trihydroxyflavone demonstrated a medium activity. This study provides for the first time a detailed screening of phenolic compounds occurring in the hydroethanolic leaf extract of E. africana. Additionally, it is shown that E. africana contains active compounds which may justify its traditional medicinal use as an antiinflammatory remedy to treat inflammatory and pain-related skin conditions in the Republic of Benin.

Les bactéries commensales digestives jouent un rôle primordial dans l’homéostasie épithéliale et la santé de l’hôte, avec notamment un rôle modulateur du système immunitaire. Des effets bénéfiques dans le traitement des pathologies inflammatoires intestinales ont été caractérisés chez certaines souches de bactéries commensales. La compréhension de ces effets dans le maintien de l’homéostasie intestinale repose sur la connaissance des interactions entre les bactéries, l’épithélium intestinal et le système immunitaire muqueux. Streptococcus salivarius est l’un des premiers colonisateurs de la cavité buccale et du tractus digestif de l’homme. Cette bactérie a été utilisée comme modèle pour rechercher des mécanismes impliquée dans l’homéostasie.La recherche d’interactions entre des souches de l’espèce S. salivarius et les cellules humaines a été réalisée pour caractériser leurs éventuelles propriétés immunomodulatrices. Nous avons montré que les bactéries vivantes et les surnageants de cultures des souches de cette espèce modulent la réponse inflammatoire in vitro via un effet inhibiteur sur l’activation de la voie NF-B dans les cellules épithéliales intestinales (HT-29 et Caco-2) et les monocytes (THP-1). Cette modulation de l’inflammation a été confirmée par la capacité des surnageants bactériens à inhiber la sécrétion d’IL-8 par les cellules épithéliales. Ces surnageants agissent via une étape impliquant IB-, un inhibiteur du facteur NF-B. Ils inhibent la dégradation de la protéine IB- phosphorylée et diminuent ainsi la translocation nucléaire des composants NF-B. Nous avons également identifié et caractérisé un métabolite bactérien présent dans ces surnageants exerçant cette activité anti-inflammatoire. L’utilisation de ce métabolite et son isomère miment in vitro l’effet inhibiteur des surnageants sur l’activation de la voie NF-B dans les cellules épithéliales et les monocytes. Nous avons ainsi caractérisé un métabolite secrété par la bactérie commensale S. salivarius qui est capable d’inhiber une des voies centrales de signalisation impliquée dans la réponse inflammatoire intestinale. Enfin, une capacité anti-inflammatoire de S. salivarius a également été montrée dans un modèle murin d’inflammation digestive dans lequel les bactéries métaboliquement actives ont protégé les animaux de colites induites avec du TNBS. Ces travaux ouvrent la voie pour le développement d’applications thérapeutiques dans le traitement de pathologies inflammatoires de l’intestin basées sur ce composé actif ou l’utilisation de S. salivarius comme probiotique. / Commensal bacteria play a vital role in epithelial homeostasis and host health, including a modulatory role of the immune system. Their beneficial effects in the treatment of inflammatory bowel disease have been characterized in some strains of commensal bacteria. Understanding these effects in maintaining intestinal homeostasis is based on the knowledge of interactions among bacteria, the intestinal epithelium and the mucosal immune system. Streptococcus salivarius is one of the first colonizers of human oral cavity and digestive tract. This bacterium was used as a template to investigate mechanisms involved in homeostasis.The research for interactions between strains of S. salivarius species and human cells was performed to characterize their possible immunomodulatory properties. We have shown that living bacteria and culture supernatants of strains of this species modulate the inflammatory response in vitro via an inhibitory effect on the activation of NF-B in intestinal epithelial cells (HT-29 and Caco-2) and monocytes (THP-1). This modulation of inflammation was confirmed by the ability of bacterial supernatants to suppress the secretion of IL-8 by epithelial cells. These supernatants act via a step involving IκB-α, an inhibitor of NF-B. They inhibit the degradation of IκB-α phosphorylated protein and thus decrease the nuclear translocation of NF-B components. We also identified and characterized a bacterial metabolite present in these supernatants exercising this anti-inflammatory activity. Use of this metabolite and its isomer in vitro mimic the repressive effect of supernatants on activation of NF-B in epithelial cells and monocytes. We have characterized a metabolite secreted by commensal bacterium S. salivarius that is capable of inhibiting one of the central signaling pathways involved in the intestinal inflammatory response. Finally, an anti-inflammatory capacity of S. salivarius was also shown in a mouse model of gastrointestinal inflammation in which the metabolically active bacteria protected the animals from colitis induced with TNBS.This work paves way for the development of therapeutic applications in the treatment of inflammatory bowel disease based on the active compound or the use of S. salivarius as a probiotic.

Les progrès réalisés en immunologie ont orienté les recherches vers des approches et des stratégies de plus en plus prometteuses et innovantes afin de mieux manipuler la réponse immunitaire. Le but de nos recherches est la prévention et le traitement des maladies liées aux dysfonctionnements du système immunitaire, telles que les maladies auto-immunes, inflammatoires et malignes. Bien que l’inflammation constitue un processus physiologique indispensable au maintien de l’homéostasie suite à une infection ou à une lésion, elle est également associée à des pathologies infectieuses, auto-immunes et tumorales. Les stratégies thérapeutiques les plus utilisées pour traiter l’inflammation sont basées sur la neutralisation des médiateurs inflammatoires par des anticorps, des antagonistes moléculaires, des immunoglobulines intraveineuses, des corticostéroïdes, des médicaments anti-inflammatoires non stéroïdiens. En plus des traitements mentionnés, des produits issus de la phytothérapie ont été largement utilisés afin d’atténuer l'inflammation et la douleur dans plusieurs maladies inflammatoires et dans le cancer. Depuis des décennies, les préparations de Viscum album, connu sous le nom de « gui européen », sont largement utilisées dans le traitement du cancer comme thérapie auxiliaire. Bien que les mécanismes d’action soient partiellement connus, plusieurs hypothèses ont été proposées. En effet, les mécanismes anti-tumoraux du Viscum album impliquent des propriétés induisant une cytotoxicité, l'apoptose, l'inhibition de l'angiogenèse et plusieurs autres mécanismes immunomodulateurs. Ce travail décrit un nouveau mécanisme anti-inflammatoire de Viscum album, qui participe à l’effet thérapeutique de ces préparations. De plus, l’effet bénéfique anti-inflammatoire observé est associé à l’inhibition des voies pro- inflammatoires de COX2 et PGE2 dans les cellules épithéliales issues d’adénocarcinome du poumon. Ce travail a identifié un des mécanismes moléculaires de Viscum album associé à son effet anti-inflammatoire participant à ses bénéfices thérapeutiques. Ainsi, ces préparations pourraient être utilisées en combinaison avec d’autres traitements dans des maladies inflammatoires et dans le cancer. / Recent advances in immunology research have led us towards more promising approaches and strategies to manipulate the immune response to prevent or treat the diseases related to immune dysfunction such as autoimmune, inflammatory pathologies and malignant diseases. Although, immuno inflammation is a basal physiological phenomenon required to eliminate the causative agent and to initiate the healing process, it is a physiopathological symptom in a diverse conditions of infectious, autoimmune and tumoral origin. Various therapeutic strategies have been developed in order to reduce inflammation and pain, including the treatment with cytokine neutralizing antibodies, molecular antagonists, intravenous immunoglobulins, corticosteroids, non-steroid anti-inflammatory drugs (NSAID) and several others. In addition to these well known anti-inflammatory therapeutic strategies, treatment with various phytotherapeutics has also contributed enormously to control inflammation and pain, associated with various severe inflammatory disorders and cancer. Viscum album (VA) preparations, commonly known as European mistletoe, are extensively used as complementary therapy in cancer for decades. However the mechanisms of action have been partially understood. Several mutually non-exclusive mechanisms have been proposed such as anti-tumor properties which involve the cytotoxic properties, induction of apoptosis, inhibition of angiogenesis and several other immunomodulatory mechanisms. This study reveals anti-inflammatory mechanism as another important mechanism of action of these phytotherapeutics, which is responsible for their therapeutic benefit and addresses the molecular mechanisms in the pro-inflammatory axis of COX-2 and PGE2 using in vitro experimental model of human lung adenocarcinoma. The present work contributes for a better understanding of mechanisms of action of Viscum album preparations underlying their therapeutic benefit and allows us to revitalize the therapeutic strategies used in treatment of inflammatory disorders and cancer.

Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2017-10-04T21:38:34Z No. of bitstreams: 1 Disserta??o - D?bora Marchesine. vers?o final.pdf: 1771951 bytes, checksum: 203519d3cb711fb7c54beb6fc48672cb (MD5) / Made available in DSpace on 2017-10-04T21:38:34Z (GMT). No. of bitstreams: 1 Disserta??o - D?bora Marchesine. vers?o final.pdf: 1771951 bytes, checksum: 203519d3cb711fb7c54beb6fc48672cb (MD5) Previous issue date: 2017-03-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Aromatic plants of the genus Lippia (Verbenaceae) are widely used in traditional medicine. A number of Lippia species can be found in the Caatinga biome in Northeastern Brazil, such as Lippia bromelyana Moldenke, endemic to the state of Bahia. This study investigated the chemical composition, acute toxicity, antinociceptive and anti-inflammatory effect of the leaf essential oil of L. bromelyana (EOLB) in mice.The chemical composition of EOLB, assessed by gas chromatography with flame ionization detection (GC/FID) and gas chromatography-mass spectrometry (GC/MS), detected limonene (29.90%) and piperitenone oxide (26.91%) as major compounds. Intraperitoneal administration of 300 mg/kg of EOLB caused no acute toxic effects or deaths in the mice. At doses of 75, 150 and 300 mg/kg caused no changes in their motor activity in the rotarod test, but promoted a significant antinociceptive effect on abdominal contortions in the acetic acid-induced constriction test, in both phases of the formalin test, glutamate or cinnamaldehyde-induced nociception tests and the Randall-Selitto test. In the hot plate test, EOLB (150 or 300 mg/kg) increased latency at 30, 60 and 120 min after administration; at a dose of 300 mg/kg, the action reversed by naloxone suggests participation of the opioid pathway. Moreover, EOLB (75, 150 or 300 mg/kg) significantly reduced the inflammatory effect of carrageenan in the paw edema test. Thus, these results suggest that EOLB promotes antinociceptive and anti-inflammatory activity by central and peripheral action. / Plantas arom?ticas do g?nero Lippia (Verbenaceae) s?o muito utilizadas pela medicina tradicional no tratamento de dist?rbios relacionados ao sistema respirat?rio e a problemas gastrointestinais. Algumas esp?cies de Lippia podem ser encontradas na caatinga, localizado na regi?o Nordeste do Brasil. Dentre essas esp?cies Lippia bromelyana Moldenke, ? end?mica do estado da Bahia. Este trabalho procurou investigar a composi??o qu?mica, o efeito t?xico agudo, antinociceptivo e anti-inflamat?rio do ?leo essencial das folhas de L. bromelyana (OELB) em camundongos. A composi??o qu?mica do OELB avaliada por CG/EM e CG/DIC, detectou limoneno (29,90%) e ?xido de piperitenona (26,91%) como compostos majorit?rios. A administra??o intraperitoneal do OELB na dose 300 mg/kg n?o causou efeitos t?xicos agudos nem mortes nos camundongos. Nas doses 75, 150 ou 300 mg/kg OELB n?o causou altera??es na atividade motora dos camundongos no teste do rota-rod, mas promoveu significante efeito antinociceptivo no teste de contor??es abdominais induzidas por ?cido ac?tico, em ambas as fases do teste da formalina, nos testes de nocicep??o induzida por glutamato ou cinamalde?do e no teste de Randall-Selitto. No teste da placa quente, OELB (150 ou 300 mg/kg) provocou o aumento na lat?ncia aos 30, 60 e 120 min ap?s sua administra??o; na dose 300 mg/kg a a??o do OELB revertida pela naloxona, sugere a participa??o da via opi?ide. Al?m disso, OELB (75, 150 ou 300 mg/kg) reduziu significativamente o efeito inflamat?rio da carragenina no teste de edema de pata. Esses resultados sugerem, portanto, que OELB promove a??o antinociceptiva e anti-inflamatoria por a??o central e perif?rica.

O hormônio alfa estimulador de melanócito (alfa-MSH) é um neuropeptídeo com atividade anti-inflamatória que apresenta efeitos benéficos em modelos experimentais de doenças autoimunes. Entretanto pouca atenção tem sido dada a seus efeitos no lúpus eritematoso sistêmico (LES). No presente estudo utilizou-se o tratamento com o super análogo NDP-MSH em modelo experimental de LES induzido por pristane. Grupos de camundongos fêmeas foram tratados diariamente com 1,25 mg/Kg de NDP-MSH ou solução salina por 180 dias. Foram avaliados os seguintes parâmetros séricos: isotipos de IgG, anticorpos antinucleares (FAN) e citocinas. A intensidade da artrite foi mensurada por graduação de edema e eritema. A função renal foi estimada por proteinúria e escore histopatológico. A expressão de IgG, alfa-SMA, iNOS, C3, CD3, MC1R, CRF e alfa-MSH glomerular foi quantificada por imunohistoquímica. Camundongos com LES apresentaram aumento de IgG, FAN, IL-6, IL-10 e TNFalfa, escore de artrite, disfunção renal e celularidade mesangial quando comparados aos animais controle normais. O tratamento dos animais LES com NDP-MSH reduziu os títulos de IgG1 e IgG2a (p < 0,05 e p < 0,001 respectivamente), bem como a incidência de FAN+ (p < 0,05). Níveis séricos de IL-6 e IL-10 foram abrandados e houve aumento dos níveis de TNFalfa (p < 0,05). O escore de artrite foi reduzido em 70% (p < 0,01). A proteinúria não foi afetada pelo tratamento, entretanto houve 50% de redução do grau de lesão glomerular (p < 0,05). A avaliação imunohistoquímica mostrou que o tratamento reduziu os depósitos de IgG e a expressão de alfa-SMA e iNOS glomerular (p < 0,01; p < 0,01 e p < 0,05 respectivamente). A expressão do receptor MC1R foi menor nos grupos LES, tratados ou não. A expressão de alfa-MSH e CRF estavam diminuídas somente no grupo tratado com NDP-MSH enquanto a expressão de C3 e CD3 não diferiu entre os grupos. Em conjunto nossos resultados sugerem, pela primeira vez, que a o tratamento com o análogo de alfa-MSH melhora a atividade da doença em modelo experimental de LES / Alpha-melanocyte stimulating hormone (alfa-MSH) is a neuropeptide with anti-inflammatory activity that has beneficial effects in experimental models of autoimmune diseases. However little attention has been paid to its effects on systemic lupus erythematosus (SLE). Herein we employed the treatment with the super analogue NDP-MSH in experimental SLE induced by pristane. Groups of female mice were treated daily with 1.25 mg/Kg of NDP-MSH (SLE-MSH) or saline for 180 days. The following parameters were evaluated: serum IgG isotypes, antinuclear antibodies (ANA) and cytokines. Arthritis was graded by edema and erythema in each paw. Renal function was estimated by proteinuria and histopathological score. The expression of IgG, alfa-SMA, iNOS, C3, CD3, MC1R, CRF and alfa-MSH was quantified by immunohistochemistry in glomerulus. SLE mice presented increased IgG levels, ANA, IL-6, IL-10 and TNFalfa, arthritis incidence, renal dysfunction and mesangial cellularity when compared with normal animals. Animals SLE-MSH presented reduction of IgG1 and IgG2a titles (p < 0.05 and p < 0.001 respectively), as well as the incidence of FAN+ (p < 0.05). The treatment reduced IL-6 and IL-10 serum levels and increase of TNFalfa levels (p < 0.05). Arthritis score was reduced in 70% in SLE-MSH animals (p < 0.01). Proteinuria was not affected by the treatment, however it was registered a 50% reduction of the glomerular lesion (p < 0.05). The immunohistochemical evaluation revealed reduced IgG deposits and glomerular expression of alfa-SMA and iNOS in SLE-MSH animals (p < 0.01, p < 0.01 and p < 0.05 vs.LES, respectively). MC1R was sub expressed in SLE animals, treated or not. The expression of alfa-MSH and CRF were impaired only in the SLE-MSH group. Taken together our results suggest for the first time that treatment with alfa-MSH analogue improve disease activity in an experimental model of SLE

O hormônio alfa estimulador de melanócito (alfa-MSH) é um neuropeptídeo com atividade anti-inflamatória que apresenta efeitos benéficos em modelos experimentais de doenças autoimunes. Entretanto pouca atenção tem sido dada a seus efeitos no lúpus eritematoso sistêmico (LES). No presente estudo utilizou-se o tratamento com o super análogo NDP-MSH em modelo experimental de LES induzido por pristane. Grupos de camundongos fêmeas foram tratados diariamente com 1,25 mg/Kg de NDP-MSH ou solução salina por 180 dias. Foram avaliados os seguintes parâmetros séricos: isotipos de IgG, anticorpos antinucleares (FAN) e citocinas. A intensidade da artrite foi mensurada por graduação de edema e eritema. A função renal foi estimada por proteinúria e escore histopatológico. A expressão de IgG, alfa-SMA, iNOS, C3, CD3, MC1R, CRF e alfa-MSH glomerular foi quantificada por imunohistoquímica. Camundongos com LES apresentaram aumento de IgG, FAN, IL-6, IL-10 e TNFalfa, escore de artrite, disfunção renal e celularidade mesangial quando comparados aos animais controle normais. O tratamento dos animais LES com NDP-MSH reduziu os títulos de IgG1 e IgG2a (p < 0,05 e p < 0,001 respectivamente), bem como a incidência de FAN+ (p < 0,05). Níveis séricos de IL-6 e IL-10 foram abrandados e houve aumento dos níveis de TNFalfa (p < 0,05). O escore de artrite foi reduzido em 70% (p < 0,01). A proteinúria não foi afetada pelo tratamento, entretanto houve 50% de redução do grau de lesão glomerular (p < 0,05). A avaliação imunohistoquímica mostrou que o tratamento reduziu os depósitos de IgG e a expressão de alfa-SMA e iNOS glomerular (p < 0,01; p < 0,01 e p < 0,05 respectivamente). A expressão do receptor MC1R foi menor nos grupos LES, tratados ou não. A expressão de alfa-MSH e CRF estavam diminuídas somente no grupo tratado com NDP-MSH enquanto a expressão de C3 e CD3 não diferiu entre os grupos. Em conjunto nossos resultados sugerem, pela primeira vez, que a o tratamento com o análogo de alfa-MSH melhora a atividade da doença em modelo experimental de LES / Alpha-melanocyte stimulating hormone (alfa-MSH) is a neuropeptide with anti-inflammatory activity that has beneficial effects in experimental models of autoimmune diseases. However little attention has been paid to its effects on systemic lupus erythematosus (SLE). Herein we employed the treatment with the super analogue NDP-MSH in experimental SLE induced by pristane. Groups of female mice were treated daily with 1.25 mg/Kg of NDP-MSH (SLE-MSH) or saline for 180 days. The following parameters were evaluated: serum IgG isotypes, antinuclear antibodies (ANA) and cytokines. Arthritis was graded by edema and erythema in each paw. Renal function was estimated by proteinuria and histopathological score. The expression of IgG, alfa-SMA, iNOS, C3, CD3, MC1R, CRF and alfa-MSH was quantified by immunohistochemistry in glomerulus. SLE mice presented increased IgG levels, ANA, IL-6, IL-10 and TNFalfa, arthritis incidence, renal dysfunction and mesangial cellularity when compared with normal animals. Animals SLE-MSH presented reduction of IgG1 and IgG2a titles (p < 0.05 and p < 0.001 respectively), as well as the incidence of FAN+ (p < 0.05). The treatment reduced IL-6 and IL-10 serum levels and increase of TNFalfa levels (p < 0.05). Arthritis score was reduced in 70% in SLE-MSH animals (p < 0.01). Proteinuria was not affected by the treatment, however it was registered a 50% reduction of the glomerular lesion (p < 0.05). The immunohistochemical evaluation revealed reduced IgG deposits and glomerular expression of alfa-SMA and iNOS in SLE-MSH animals (p < 0.01, p < 0.01 and p < 0.05 vs.LES, respectively). MC1R was sub expressed in SLE animals, treated or not. The expression of alfa-MSH and CRF were impaired only in the SLE-MSH group. Taken together our results suggest for the first time that treatment with alfa-MSH analogue improve disease activity in an experimental model of SLE