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H9c2(2-1)-based sulforhodamine B assay as a possible alternative in vitro platform to investigate effluent and metals toxicity on fish
Chemosphere ( IF 8.8 ) Pub Date : 2021-02-24 , DOI: 10.1016/j.chemosphere.2021.130009
Elsa T. Rodrigues , Miguel A. Pardal , Eduarda Pereira , Joana F. Monteiro , Ana C. Certal , Paulo J. Oliveira

To overcome restrictions on the use of fish in toxicity testing, the present study proposes to compare the 50% growth inhibition potential (EC50) of four types of effluents on the rat cardiomyoblast H9c2(2-1) cell line by using the sulforhodamine B (SRB) cell mass colorimetric assay, with the corresponding fish lethal test results. Our objective was to evaluate if H9c2(2-1) cells shows comparable sensitivities, in both relative and absolute terms, to those provided by fish. In parallel, this study also compared the results of the chemical characterization with the legislation in force for environmental protection against effluent release into the receiving environment. Moreover, we tested the H9c2(2-1)-based SRB assays with the metals of concern found in the effluent samples. Both fish and cell assays showed the same toxicity rank for effluents: Metal > Oil > Municipal > Paper, and it should be stressed that the complementarity of using chemical and biological data represents a step forward to guarantee both environmental and human safety, since the chemical characterization showed a different toxicity rank: Metal > Municipal > Oil > Paper. Regarding metal elements, the short-term fish results showed a toxicity rank non-comparable with the rank obtained for cells. Nevertheless, the gathered results reveal the potentiality of the in vitro H9c2(2-1) platform as an alternative for fish lethal testing to assess, in absolute terms, the toxicity of effluents, particularly municipal effluents, and metals.



中文翻译:

基于H9c2(2-1)的磺基罗丹明B测定法可能是替代的体外平台,可用于研究废水和金属对鱼类的毒性

为了克服毒性试验中鱼类使用的限制,本研究建议比较50%生长抑制潜力(EC 50)通过使用磺基罗丹明B(SRB)细胞质量比色法测定大鼠心肌成纤维细胞H9c2(2-1)细胞系上的四种类型的废水,并获得相应的鱼致死性测试结果。我们的目标是评估H9c2(2-1)细胞是否在相对和绝对方面都可与鱼类提供的敏感性相媲美。同时,本研究还比较了化学表征的结果和现行的环境保护立法,以防止废水排放到接收环境中。此外,我们用废水样品中发现的有关金属测试了基于H9c2(2-1)的SRB分析。鱼和细胞分析均显示出相同的污水毒性等级:金属 > 石油 > 市政 > 纸张,应该强调的是,使用化学和生物学数据的互补性代表了向保证环境和人类安全的方向迈出的一步,因为化学表征显示出不同的毒性等级:金属 > 市政 > 石油 > 纸张。关于金属元素,短期鱼类结果显示其毒性等级与细胞获得的等级无法比拟。但是,收集的结果表明,体外H9c2(2-1)平台可作为鱼类致死性测试的替代品,以绝对值评估废水(尤其是市政废水和金属)的毒性。

更新日期:2021-02-28
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