Cytotoxicity of settling particulate matter and sediments of the Neckar River (Germany) during a winter flood
Cytotoxicity of settling particulate matter and sediments of the Neckar River (Germany) during a winter flood
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DOI：
10.1002/etc.5620190302
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发表时间：
2000-03-01
影响因子：
4.1
通讯作者：
Braunbeck, T
中科院分区：
文献类型：
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作者：
Hollert, H;Dürr, M;Braunbeck, T
为了研究内卡河（中欧一条锁定调节的河流）携带的沉淀颗粒物（SPM）的细胞毒性和基因毒性潜力，在一场严重的急性细胞毒性战争中，使用成纤维细胞样鱼细胞系RTG-2进行了研究，以中性红色保留、琥珀酸脱氢酶（MTT）和乳酸脱氢酶（LDH）释放试验以及显微镜检查为终点。采用Ames试验评估水、孔隙水、沉积物和SPM的遗传毒性。不同的提取方法（可变极性溶剂索氏提取以及根据pH值的流体/流体提取）以及补充生物试验，从-萘-黄酮/苯巴比妥诱导的大鼠肝脏中提取S9馏分，可以进一步表征生物损伤。沉积物和SPM提取物均对RTG-2细胞有细胞毒性作用。细胞毒性随着提取溶剂的极性而显著增加（NR50 =中性红试验中50%细胞死亡的有效浓度：乙醇、丙酮、二氯甲烷和正己烷提取物分别为80[65]、100[70]、180[220]和225 [270]mg/ml，如果添加[未添加]S9进行测量）。根据pH提取后，细胞毒性主要归因于中性物质（中性部分和酸性部分分别为NR50: 80和218 mg干SPM/ml试验介质），而微酸和碱性部分已经表现出很少或没有细胞毒性。胡德上升期间的样品显示出最高的细胞毒活性。添加S9制剂可显著增强细胞毒性。相比之下，在天然地表水、孔隙水和SPM中没有发现遗传毒性活性。
To investigate the cytotoxic and genotoxic potentials of settling particulate matter (SPM) carried by the Neckar River, a well-studied model for a lock-regulated river in central Europe, during a hood, acute cytotoxicity war investigated using the fibroblast-like fish cell line RTG-2 with the neutral red retention, the succinic acid dehydrogenase (MTT), and the lactatedehydrogenase (LDH) release assays as well as microscopic inspection as endpoints. Genotoxicity of water, pore water, sediments, and SPM were assessed using the Ames test. Different extraction methods (Soxhlet extraction with solvents of variable polarity as well as a fluid/fluid extraction according to pH) in addition to a supplementation of biotests with S9 fractions from the liver of beta-naphthoflavone/phenobarbital-induced rats allowed a further characterization of the biological damage. Both sediments and SPM extracts caused cytotoxic effects in RTG-2 cells. Cytotoxicity was found to increase significantly with polarity of extracting solvents (NR50 = effective concentration for 50% cell death in the neutral red test: 80 [65], 100 [70], 180 [220], and 225 [270] mg/ml for ethanol, acetone, dichloromethane, and n-hexane extracts, respectively, if measured with [without] S9 supplementation). Following extraction according to pH, cytotoxicity could be attributed mainly to neutral substances (NR50: 80 and 218 mg dry SPM/ml test medium for the neutral and the acid fractions, respectively), whereas the slightly acid and basic fractions already showed little or no cytotoxicity. Samples taken during the period of Hood rise showed the highest cytotoxic activities. Cytotoxicity was significantly enhanced by the addition of S9 preparations. In contrast, no genotoxic activity was found in native surface waters, pore waters, and SPM.