Department of Biology, University of Milan, Milan, Italy.
Ecotoxicology. 2012 Mar;21(2):379-92. doi: 10.1007/s10646-011-0799-6. Epub 2011 Oct 5.
Non-steroidal anti-inflammatory drugs (NSAIDs) are the sixth top-selling drugs worldwide and are commonly found in freshwater ecosystems in the high ng/l to low μg/l range. Recent studies have investigated both the acute and the chronic toxicity of single NSAIDs on different biological models, but these studies have completely neglected the fact that, in the environment, non-target organisms are exposed to mixtures of drugs that have unforeseeable toxicological behavior. This work investigated the sub-lethal effects induced by a mixture of three common NSAIDs, namely, diclofenac, ibuprofen and paracetamol, on the freshwater bivalve, the zebra mussel (Dreissena polymorpha). The mussels were exposed to three different environmental concentrations of the mixture (Low, Mid and High). A multi-biomarker approach was used to highlight cyto-genotoxic effects and the imbalance of the oxidative status of the treated specimens. The Neutral Red Retention Assay (NRRA) was used as a biomarker of cytotoxicity, whereas the activities of catalase, superoxide dismutase, glutathione peroxidase and glutathione S-transferase were measured to assess the role played by the oxidative stress enzymes. In addition, the single cell gel electrophoresis assay, the DNA Diffusion assay and the micronucleus test were used to investigate possible genotoxic effects. According to our NRRA results, each treatment was able to induce a significant cellular stress in bivalves, probably due to the raise of oxidative stress, as indicated by the alteration of enzyme activities measured in treated specimens. Moreover, the mixture induced significant enhancements of DNA fragmentation, which preluded fixed genetic damage, as highlighted by the increase of both apoptotic and micronucleated cells.
非甾体抗炎药（NSAIDs）是全球销量第六的药物，在高 ng/L 到低 μg/L 的范围内普遍存在于淡水生态系统中。最近的研究调查了单一 NSAIDs 对不同生物模型的急性和慢性毒性，但这些研究完全忽略了一个事实，即在环境中，非靶标生物会暴露于具有不可预测的毒理学行为的药物混合物中。本工作研究了三种常见 NSAIDs（双氯芬酸、布洛芬和扑热息痛）混合物对淡水双壳贝类斑马贻贝（Dreissena polymorpha）的亚致死效应。贻贝暴露于混合物的三种不同环境浓度（低、中和高）中。采用多生物标志物方法来突出细胞毒性和处理标本氧化状态失衡的影响。中性红保留试验（NRRA）被用作细胞毒性的生物标志物，而过氧化氢酶、超氧化物歧化酶、谷胱甘肽过氧化物酶和谷胱甘肽 S-转移酶的活性则用于评估氧化应激酶所起的作用。此外，单细胞凝胶电泳试验、DNA 扩散试验和微核试验用于研究可能的遗传毒性效应。根据我们的 NRRA 结果，每种处理都能在双壳贝类中诱导出明显的细胞应激，这可能是由于氧化应激的增加，这是通过测量处理标本中的酶活性变化来表明的。此外，该混合物诱导了 DNA 片段化的显著增强，预示着固定遗传损伤，这一点可以从凋亡和微核细胞的增加中得到突出体现。
