Cellular responses of Prochilodus lineatus hepatocytes after cylindrospermopsin exposure |
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Authors: | S Liebel CA Oliveira Ribeiro RC Silva WA Ramsdorf MM Cestari VF Magalhães JRE Garcia BM Esquivel F Filipak Neto |
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Institution: | aDepartamento de Biologia Celular, Universidade Federal do Paraná, Caixa Postal 19031, CEP 81.531-990 Curitiba, PR, Brazil;bDepartamento de Genética, Universidade Federal do Paraná, Caixa Postal 19031, CEP 81.531-990 Curitiba, PR, Brazil;cInstituto de Biofísica Carlos Chagas Filho, Centro de Ciências da Saúde, Bloco G, Ilha do Fundão, Universidade Federal do Rio de Janeiro, CEP 21949-900 Rio de Janeiro, RJ, Brazil;dEstação de Piscicultura Panamá, Paulo Lopes, SC, Brazil;eCampus Laranjeiras do Sul, Universidade Federal da Fronteira Sul, CEP 85303-775 Laranjeiras do Sul, PR, Brazil |
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Abstract: | Cylindrospermopsin is a potent toxicant for eukaryotic cells produced by several cyanobacteria. Recently, primary hepatocyte cultures of Neotropical fish have been established, demonstrating to be a quite efficient in vitro model for cellular toxicology studies. In the current study, a protocol for culture of Prochilodus lineatus hepatocytes was established and utilized to investigate the cellular responses to purified cylindrospermopsin exposure. Hepatocytes were successfully dissociated with dispase, resulting in a cell yield of 6.36 × 107 cells g−1 of liver, viability of 97% and attachment on uncoated culture flasks. For investigation of cylindrospermopsin effects, hepatocytes were dissociated, cultured during 96 h and exposed to three concentrations of the toxin (0.1, 1.0 or 10 μg l−1) for 72 h. Cylindrospermopsin exposure significantly decreased cell viability (0.1 and 1 μg l−1) and multixenobiotic resistance mechanism, MXR (all exposed groups), but increased reactive oxygen/nitrogen species levels (all exposed groups) and lipid peroxidation (10 μg l−1). On the other hand no significant alterations were observed for other biochemical biomarkers as 2GSH/GSSG ratio, protein carbonyl levels and DNA strand breaks or glutathione S-transferase and glucose 6-phosphate dehydrogenase activities. In conclusion, hepatocytes might be made sensitive to cylindrospermopsin, at least in part, due to reduction of xenobiotics and endobiotics efflux capacity by MXR. Additionally, the toxin exposure suggests important issues regarding hepatocytes survival at the lowest cylindrospermopsin concentrations. |
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Keywords: | Cylindrospermopsin Hepatocytes primary culture Prochilodus lineatus Multixenobiotic resistance Oxidative stress |
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