Carbendazim induz morte em células epiteliais alveolares: cultura celular submersa e em interface ar-líquido
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2019-03-21
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Universidade Federal de Goiás
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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.
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TOLLSTADIUS, Bruna Ferreira. Carbendazim induz morte em células epiteliais alveolares: cultura celular submersa e em interface ar-líquido. 2019. 60 f. Dissertação (Mestrado em Ciências Farmacêuticas) - Universidade Federal de Goiás, Goiânia, 2019.