Bianchi Jaqueline, Cabral-de-Mello Diogo Cavalcanti, Marin-Morales Maria Aparecida
Department of Biology, Institute of Biosciences, São Paulo State University (UNESP), Av. 24A, 1515, Bela Vista, Rio Claro, São Paulo CEP 13506-900, Brazil.
Department of Biology, Institute of Biosciences, São Paulo State University (UNESP), Av. 24A, 1515, Bela Vista, Rio Claro, São Paulo CEP 13506-900, Brazil.
Ecotoxicol Environ Saf. 2015 Oct;120:174-83. doi: 10.1016/j.ecoenv.2015.05.040. Epub 2015 Jun 12.
The insecticide imidacloprid and the herbicide sulfentrazone are two different classes of pesticides that are used for pest control in sugarcane agriculture. To evaluate the genotoxic potential of low concentrations of these two pesticides alone and in mixture, the comet assay and the micronucleus (MN) test employing fluorescence in situ hybridization (FISH) with a centromeric probe were applied in human hepatoma cell lines (HepG2), in a 24-h assay. Mutagenicity was assessed by Salmonella/microsome assay with TA98 and TA100 strains in the absence and presence of an exogenous metabolizing system (S9). The results showed significant inductions of MN in HepG2 cells by both pesticides, for all the tested concentrations. As evidenced in the comet assay, only the imidacloprid presented significant responses. When the two pesticides were associated, a significant induction of damage was observed in the HepG2 cells by the comet assay, but not by the MN test. Moreover, the MN induced by the mixtures of the pesticides appeared at lower levels than those induced by sulfentrazone and imidacloprid when tested alone. According to the FISH results, the damage induced by imidacloprid in the HepG2 cells resulted from a clastogenic action of this insecticide (76.6% of the MN did not present a centromeric signal). For the herbicide sulfentrazone and for the mixture of the pesticides, a similar frequency of MN with and without the presence of the centromeric signal (herbicide: 52.45% of the MN without centromeric signal and 47.54% of the MN with centromeric signal; mixture: 48.71% of the MN without centromeric signal and 51.42% of the MN with centromeric signal) was verified. Based on these results, it was concluded that each one of the pesticides evaluated interacts with the DNA of HepG2 cells and causes irreparable alterations in the cells. However, the combination of the pesticides showed an antagonistic effect on the cells and the damage induced was milder and not persistent in HepG2 cells. The results obtained by the Ames test did not point out significant results.
杀虫剂吡虫啉和除草剂乙磺隆是用于甘蔗种植中害虫防治的两类不同农药。为评估低浓度的这两种农药单独及混合使用时的遗传毒性潜力，在人肝癌细胞系（HepG2）中进行了24小时试验，采用彗星试验和使用着丝粒探针的荧光原位杂交（FISH）微核（MN）试验。在有无外源性代谢系统（S9）的情况下，用TA98和TA100菌株通过沙门氏菌/微粒体试验评估致突变性。结果显示，对于所有测试浓度，两种农药均在HepG2细胞中显著诱导微核形成。如彗星试验所示，只有吡虫啉呈现出显著反应。当两种农药联合使用时，彗星试验显示HepG2细胞中损伤显著诱导，但微核试验未显示。此外，与单独测试时相比，农药混合物诱导的微核水平低于乙磺隆和吡虫啉诱导的微核水平。根据FISH结果，吡虫啉在HepG2细胞中诱导的损伤是由该杀虫剂的断裂作用引起的（76.6%的微核未呈现着丝粒信号）。对于除草剂乙磺隆和农药混合物，有和没有着丝粒信号的微核频率相似（除草剂：52.45%的微核无着丝粒信号，47.54%的微核有着丝粒信号；混合物：48.71%的微核无着丝粒信号，51.42%的微核有着丝粒信号）。基于这些结果得出结论，所评估的每种农药均与HepG2细胞的DNA相互作用并导致细胞中不可修复的改变。然而，农药组合对细胞显示出拮抗作用，且在HepG2细胞中诱导的损伤较轻且不持久。艾姆斯试验获得的结果未显示出显著结果。
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