Carbendazim induz morte em células epiteliais alveolares: cultura celular submersa e em interface ar-líquido

• Ano de defesa: 2019
• Autor(a) principal: Tollstadius, Bruna Ferreira
• Orientador(a): Valadares, Marize Campos
• Banca de defesa: Valadares, Marize Campos, Biancardi , Manoel Francisco, Silva , Luís Antônio Dantas da, Oliveira , Gisele Augusto Rodrigues de
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Goiás
• Programa de Pós-Graduação: Programa de Pós-graduação em Ciências Farmacêuticas (FF)
• Departamento: Faculdade Farmácia - FF (RG)
• País: Brasil
• Palavras-chave em Português: Carbendazim; Toxicidade inalatória; Modelo epitelial alveolar 3D; Exposição por nebulização; Interface ar-líquido
• Palavras-chave em Inglês: Inhalation toxicity; Alveolar 3D epithelial model; Nebulization exposure; Air-liquid interface
• Área do conhecimento CNPq: FARMACOLOGIA::TOXICOLOGIA
• Link de acesso: http://repositorio.bc.ufg.br/tede/handle/tede/10345

Introduction: The fungicide Carbendazim is widely used in agriculture and preservation of films and fibers. It’s well stablished that in mammals it can promote germ cell mutagenicity, carcinogenicity, and reproductive toxicity. However, few data about the effects of this toxicant upon the respiratory system are available. Objectives: Evaluate Carbendazim toxicity upon A549 alveolar cells in monolayer, as well as on a reconstructed alveolar epithelial model using 3D culture strategy. Methodology: Carbendazim cytotoxicity upon A549 cells was assessed through MTT reduction method. For mechanistic studies, also performed on monolayer, cells were exposed to the fungicide (12.5µM) for 24 hours, analyzed by flow cytometry, being investigated for cell cycle progression, ROS generation, and apoptosis potential induction. For 3D model obtention, A549 cell were cultivated on transwell inserts, upon collagen type I matrix, on an air-liquid interface for two days. The 3D model was then characterized usingindirect immunofluorescence, and evaluated for citokeratin, CD44, MUC-1, α-tubulin and E- cadherine expression, which showing a similar expression pattern to the human alveoli. After characterized, this model was exposed to Carbendazim (12.5µM) on the exposure chamber with aerosol production, and assessed for Nrf2, Cleaved Caspase-3 and α-tubuline expression by indirect immunofluorescence, as well as the mitochondrial activity and ROS generation.