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Paula Rocha Moreirahttp://lattes.cnpq.br/5116898796555782http://lattes.cnpq.br/6262402481877575Simone Angélica de Faria Amormino Carvalho2024-03-08T18:46:58Z2024-03-08T18:46:58Z2016-07-12http://hdl.handle.net/1843/65567A metilação do DNA é um mecanismo epigenético catalisado por uma família de enzimas denominadas DNA metiltransferases (DNMTs). Este evento é capaz de interferir na expressão de vários genes envolvidos na resposta imune e tem sido relacionado à patogênese de doenças inflamatórias, como a periodontite. CD14 é um receptor com alta afinidade pelo LPS e ao se ligar a ele, promove uma cascata de sinalização intracelular com ativação do NFkB e consequente expressão de genes próinflamatórios. Os objetivos do presente trabalho foram avaliar o padrão de metilação e a transcrição dos genes CD14 e NFkB, assim como a transcrição das enzimas DNMTs na periodontite crônica. A influência do tabagismo foi avaliada nestes eventos. Amostras gengivais foram coletadas de 30 indivíduos com periodontite crônica e 30 sem periodontite e submetidas à análise histológica. O DNA foi extraído pelo kitQIAamp DNA Tissue Mini Kit e analisado utilizando o Methyl Profiler DNA Methylation qPCR, com posterior análise por PCR em tempo real. Para a avaliação da transcrição, foi realizada a extração do RNA pelo método do trizol, seguida da conversão em cDNA e posterior análise por PCR em tempo real com sondas específicas. Um padrão de DNA não metilado do CD14 foi observado em todos os grupos analisados. A transcrição de CD14 foi maior no grupo periodontite comparado ao controle. Um padrão de DNA não metilado do NFkB também foi um evento usual nos tecidos gengivais dos indivíduos com e sem periodontite. Maior transcrição de NFkB foi verificada no grupo sem periodontite comparado ao com periodontite, considerando os não fumantes. Não houve diferença na transcrição da DNMT1 em todos grupos avaliados. Os indivíduos com periodontite crônica e fumante apresentaram maior expressão da DNMT3A e DNMT3B que os não fumantes. DNMT3A e B foram correlacionadas com parâmetros de gravidade da doença periodontal. Os resultados mostraram pela primeira vez que o padrão hipometilado dos genes CD14 e NFkB é um evento comum nos tecidos periodontais. Além disso, os dados sugerem o fumo pode interferir na transcrição de CD14 e das DNMT3A e B. Estudos adicionais são necessários para determinar a relevância funcional dessas alterações na periodontite.DNA methylation is an epigenetic mechanism catalyzed by a family of enzymes called DNA methyltransferases (DNMTs). This event is able to interfere with the expression of several genes involved in immune response and has been related to the pathogenesis of inflammatory diseases, like periodontitis. CD14 is a receptor with high affinity for LPS and this interaction promotes a cascade of intracellular signaling with NFkB activation and consequent expression of pro-inflammatory genes.The objectives of this study were to assess the pattern of methylation and transcription of CD14 and NFkB genes, as well as the transcription of DNMT on chronic periodontitis.The influenceof smoking was evaluated in these events.Gingival tissue samples were collected from 30 individuals with chronic periodontitis and 30 without periodontitis and submitted to histological analysis.The DNA was extracted by the QIAamp DNA Tissue kit Mini Kit and analyzed using the Methyl DNA Methylation qPCR Profiler. For the evaluation of transcription RNA extraction was performed by the method of trizol, followed by conversion to cDNA and further analysis by real-time PCR with specific probes. An unmethylated pattern was observed in all groups. The CD14 transcription was higher in periodontitis compared to control. The unmethylation of NFkB was also a usual event in gingival tissues from individuals with and without periodontitis. Increased transcription of NFkB was observed in the group without periodontitis compared to with periodontitis, considering the non-smokers. There was no difference in transcription of DNMT1 in all evaluated groups. Individuals with chronic periodontitis and smoker presented a higher expression of DNMT3A and DNMT3B than nonsmokers. DNMT3A and B were correlated with severity of periodontal disease.The results showed for the first time a unmethylated patternof the CD14 and NFkB genes as a common event in periodontal tissues. In addition the data suggest that the smoking can interfere in transcription of CD14 and DNMT3A/B. Additional studies are needed to determine the functional relevance of these changes on periodontitis.porUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em Biologia CelularUFMGBrasilhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessPeriodontiteMetiltransferasesDNAReceptores de LipopolissacarídeosPeriodontiteDNADNA metiltransferases (DNMTs)CD14NFkBAvaliação do padrão de metilação dos genes CD14 e NFkB e transcrição das DNAs metiltransferases na Periodontite Crônicainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALTese doutorado Simone Angélica.pdfTese doutorado Simone Angélica.pdfapplication/pdf2002046https://repositorio.ufmg.br/bitstream/1843/65567/1/Tese%20doutorado%20Simone%20Ang%c3%a9lica.pdf447aab8c112306ba7758145f15dd31c5MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufmg.br/bitstream/1843/65567/2/license_rdfcfd6801dba008cb6adbd9838b81582abMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/65567/3/license.txtcda590c95a0b51b4d15f60c9642ca272MD531843/655672024-03-08 15:46:59.411oai:repositorio.ufmg.br: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ório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2024-03-08T18:46:59Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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