Universidade Federal do Rio Grande-FURG, Instituto de Ciências Biológicas (ICB), Campus Carreiros, Av. Itália km 8 s/n (96200-970), Rio Grande, RS, Brazil; Programa de Pós Graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas (ICB), FURG, Brazil; Rede de Nanotoxicologia (MCTI/CNPq), Nanotoxicologia ocupacional e ambiental: subsídios científicos para estabelecer marcos regulatórios e avaliação de riscos, Rio Grande, RS, Brazil.
Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Buenos Aires, Argentina.
Aquat Toxicol. 2014 Feb;147:76-83. doi: 10.1016/j.aquatox.2013.12.007. Epub 2013 Dec 14.
Compounds from the nanotechnology industry, such as carbon-based nanomaterials, are strong candidates to contaminate aquatic environments because their production and disposal have exponentially grown in a few years. Previous evidence shows that fullerene C60, a carbon nanomaterial, can facilitate the intake of metals or PAHs both in vivo and in vitro, potentially amplifying the deleterious effects of these toxicants in organisms. The present work aimed to investigate the effects of fullerene C60 in a Danio rerio (zebrafish) hepatocyte cell lineage exposed to benzo[a]pyrene (BaP) in terms of cell viability, oxidative stress parameters and BaP intracellular accumulation. Additionally, a computational docking was performed to investigate the interaction of the fullerene C60 molecule with the detoxificatory and antioxidant enzyme πGST. Fullerene C60 provoked a significant (p<0.05) loss in cellular viability when co-exposed with BaP at 0.01, 0.1 and 1.0 μg/L, and induced an increase (p<0.05) in BaP accumulation in the cells after 3 and 4h of exposure. The levels of reactive oxygen species (ROS) in the cells exposed to BaP were diminished (p<0.05) by the fullerene addition, and the increase of the GST activity observed in the BaP-only treated cells was reduced to the basal levels by co-exposure to fullerene. However, despite the potential of the fullerene molecule to inhibit π GST activity, demonstrated by the computational docking, the nanomaterial did not significantly (p>0.05) alter the enzyme activity when added to GST purified extracts from the zebrafish hepatocyte cells. These results show that fullerene C60 can increase the intake of BaP into the cells, decreasing cell viability and impairing the detoxificatory response by phase II enzymes, such as GST, and this latter effect should be occurring at the transcriptional level.
来自纳米技术行业的化合物，如基于碳的纳米材料，是污染水生环境的强有力候选物，因为它们的生产和处置在短短几年内呈指数级增长。先前的证据表明，富勒烯 C60 是一种碳纳米材料，它可以促进金属或多环芳烃在体内和体外的摄入，从而可能放大这些有毒物质对生物体的有害影响。本研究旨在研究富勒烯 C60 在斑马鱼（斑马鱼）肝细胞系中暴露于苯并[a]芘（BaP）时对细胞活力、氧化应激参数和 BaP 细胞内积累的影响。此外，还进行了计算对接，以研究富勒烯 C60 分子与解毒和抗氧化酶π GST 的相互作用。当富勒烯 C60 与 0.01、0.1 和 1.0μg/L 的 BaP 共同暴露时，会显著（p<0.05）降低细胞活力，并在暴露 3 和 4 小时后诱导细胞内 BaP 积累增加（p<0.05）。暴露于 BaP 的细胞中的活性氧（ROS）水平（p<0.05）因富勒烯的加入而降低，而仅用 BaP 处理的细胞中 GST 活性的增加则因与富勒烯共同暴露而降低到基础水平。然而，尽管计算对接表明富勒烯分子具有抑制π GST 活性的潜力，但当将其添加到来自斑马鱼肝细胞的 GST 纯化提取物中时，纳米材料并未显著（p>0.05）改变酶活性。这些结果表明，富勒烯 C60 可以增加 BaP 进入细胞的摄取，降低细胞活力，并通过 GST 等 II 相酶损害解毒反应，而后者的影响可能发生在转录水平。
