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Fabíola Mara Ribeirohttp://lattes.cnpq.br/8721252207572060Vinícius de Toledo RibasAllysson Thiago Cramer Soareshttp://lattes.cnpq.br/1266795827093960Nathália Costa Silva2023-05-15T16:07:53Z2023-05-15T16:07:53Z2022-07-21http://hdl.handle.net/1843/53371A esclerose múltipla (EM) é uma doença neuroinflamatória, autoimune, crônica e incapacitante, em que se desenvolve uma resposta imune auto-reativa voltada para proteínas da bainha de mielina, bem como infiltrado inflamatório no SNC, excitotoxicidade, astrogliose e neurodegeneração. Os astrócitos, que constituem o tipo celular mais abundante no SNC de mamíferos, se tornam reativos mediante injúria. Já foi demonstrado que a ativação do receptor metabotrópico de glutamato 5 (mGluR5) é neuroprotetora mediante insulto de glutamato e pode inibir a sinalização microglial pró-inflamatória in vitro, bem como induzir a expressão microglial do fator neurotrófico derivado do cérebro (BDNF) em modelo murino da EM. Atualmente, não existem abordagens terapêuticas capazes de tratar de modo simultâneo a neuroinflamação e a neurodegeneração que ocorrem durante a EM. No presente trabalho, buscou-se estabelecer estímulo pró-inflamatório com as citocinas associadas à patologia da EM e à reatividade astrocitária, fator de necrose tumoral alfa (TNF-α), interleucina 1 beta (IL-1β) e fator estimulador de colônias de granulócitos e macrófagos (GM-CSF), e avaliar possível efeito protetor do modulador alostérico positivo (MAP) do mGluR5, o VU0409551, em culturas de astrócitos derivadas de células tronco humanas de pluripotência induzida (hiPSCs) submetidas ao insulto pró-inflamatório. Observamos aumento da expressão de TNF-α e IL-1β a partir do estímulo com TNF-α e IL-1β por 24h. Além disso, o VU0409551 induziu aumento da expressão de IL-6, de modo independente do estímulo com TNF-α e IL- 1β. Observou-se também efeito marginal do tratamento com 1μM de VU0409551 na prevenção do aumento da expresão de IL-1β induzida pelo estímulo pró-inflamatório com TNF-α e IL-1β ao longo de 24h. Os resultados não sugerem alterações na expressão dos demais alvos avaliados, IL-10, GNR, BDNF e EAAT1, e nas concentrações de glutamato dos meios condicionados de astrócitos estimulados com TNF-α, IL-1β e tratados com VU0409551. Desse modo, o possível potencial anti- infamatório de VU0409551 não foi confirmado pelos resultados no presente trabalho.Multiple sclerosis (MS) is a neuroinflammatory, autoimmune, chronic and disabling disease that affects the central nervous system due to an autoimmune response against myelin proteins, along with the formation of inflammatory infiltrate, demyelination, excitotoxicity, astrogliosis and neurodegeneration. Astrocytes are the most abundant cell type in the mammalian central nervous system (CNS) and, upon injury, they become reactive. Activation of metabotropic glutamate receptor subtype 5 (mGluR5) has been shown to be neuroprotective upon glutamate insult and can inhibit pro-inflammatory microglial signaling in vitro, as well as induce microglial and astrocytic BDNF expression in a murine model of MS. Currently, there are no therapeutic approache that could mitigate both MS-related neuroinflamation and neurogeneration. In the present study, we aimed to establish a pro-inflammatory stimulus with cytokines associated with MS pathology and astrocytic reactivity, tumor necrosis factor alpha (TNF -α), interleukin 1 beta (IL-1β) and granulocyte and macrophage colony stimulating factor (GM-CSF) and to evaluate the effect of the positive modulator (PAM) of mGluR551, VU0409551, in stimulated astrocytes derived from human induced pluripotency cells (hiPSCs). We observed an increase in the expression of TNF-α and IL-1β 24 hours following TNF-α and IL-1β stimulation. VU0409551 was found to increase the expression of IL-6, regardless of TNF-α and IL-1β stimulation. Furthermore, a marginal effect of treatment with 1μM of VU0409551 was observed in the prevention of the elevation of IL-1β expression induced by the pro-inflammatory stimulus with TNF-α and IL-1β over 24h. The results indicate that TNF-α, IL-1β and VU0409551 treatment does not change the expression of the other evaluated targets IL-10, GNR, BDNF, EAAT1, and does not modify glutamate levels in astrocyte conditioned media. Therefore, the assumed anti- inflammatory effect of VU0409551 was not confirmed by the results in this study.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorporUniversidade Federal de Minas GeraisCurso de Especialização em Bioquímica e ImunologiaUFMGBrasilICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIABioquímica e imunologiaEsclerose MúltiplaAstrócitosReceptor de Glutamato MetabotrópicoEsclerose Múltiplareatividade astrocitáriamGluR5PAMVU0409551Avaliação do tratamento com VU0409551 em astrócitos derivados de hiPSCs estimulados com citocinas pró-inflamatórias associadas à Esclerose Múltiplainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALDissertação Bioquímica e Imunologia Nathália Costa Silva 07.2022.pdfDissertação Bioquímica e Imunologia Nathália Costa Silva 07.2022.pdfapplication/pdf1915482https://repositorio.ufmg.br/bitstream/1843/53371/3/Disserta%c3%a7%c3%a3o%20Bioqu%c3%admica%20e%20Imunologia%20Nath%c3%a1lia%20Costa%20Silva%2007.2022.pdf55458dd6f00c3df7a55dc9701e931082MD53LICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/53371/4/license.txtcda590c95a0b51b4d15f60c9642ca272MD541843/533712023-05-15 13:07:54.151oai:repositorio.ufmg.br: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ório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-05-15T16:07:54Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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