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通过生化和流式细胞术检测方法确定农达经典款及其成分对NE - 4C和MC3T3 - E1细胞系的细胞毒性作用。

Cytotoxic effects of Roundup Classic and its components on NE-4C and MC3T3-E1 cell lines determined by biochemical and flow cytometric assays.

作者信息

Oláh Marianna, Farkas Enikő, Székács Inna, Horvath Robert, Székács András

机构信息

Agro-Environmental Research Centre, Institute of Environmental Sciences, Hungarian University of Agriculture and Life Sciences, Herman Ottó u. 15, H-1022 Budapest, Hungary.

Nanobiosensorics Laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research, Konkoly-Thege M. u. 29-33, H-1121 Budapest, Hungary.

出版信息

Toxicol Rep. 2022 Apr 20;9:914-926. doi: 10.1016/j.toxrep.2022.04.014. eCollection 2022.

Abstract

Cytotoxic effects of the market leading broad-spectrum, synthetic herbicide product Roundup Classic, its active ingredient glyphosate (in a form of its isopropylamine (IPA) salt) and its formulating surfactant polyethoxylated tallowamine (POE-15) were determined on two murine cell lines, a neuroectodermal stem cell-like (NE-4C) and a high alkaline phosphatase activity osteoblastic cell line (MC3T3-E1). Cytotoxicity, genotoxicity, effects on cell viability and cell cycles were examined in five flow cytometry tests, the two former of which were compared by the enzymatic-assay and the alkaline single cell gel electrophoresis (Comet) assay. All of the tests indicated the NE-4C cells being more sensitive, than the MC3T3-E1 cell line to the treatments with the target compounds. Higher sensitivity differences were detected in the viability test by flow cytometry (7-9-fold), than by the MTT assay (1.5-3-fold); in the genotoxicity test by the Comet assay (3.5-403-fold), than by the DNA-damage test (9.3-158-fold); and in the apoptosis test by the Annexin V dead cell kit (1.1-12.7-fold), than by the Caspase 3/7 kit (1-6.5-fold). Cell cycle assays indicated high count of cells (70%) in the G0/G1 phase for MC3T3-E1 cells, than in NE-4C cell (40%) after 24 h. The order of the inhibitory potency of the target substances has unequivocally been POE-15 > Roundup Classic > > glyphosate IPA salt.

摘要

测定了市场领先的广谱合成除草剂产品农达经典、其活性成分草甘膦(异丙胺(IPA)盐形式)及其配方表面活性剂聚乙氧基化牛脂胺(POE - 15)对两种小鼠细胞系的细胞毒性,即神经外胚层干细胞样细胞系(NE - 4C)和高碱性磷酸酶活性成骨细胞系(MC3T3 - E1)。通过五项流式细胞术检测来检查细胞毒性、遗传毒性、对细胞活力和细胞周期的影响,其中前两项通过酶促测定和碱性单细胞凝胶电泳(彗星试验)进行比较。所有测试均表明,NE - 4C细胞比MC3T3 - E1细胞系对目标化合物处理更敏感。在流式细胞术活力测试中检测到的敏感性差异(7 - 9倍)高于MTT测定(1.5 - 3倍);在彗星试验遗传毒性测试中(3.5 - 403倍)高于DNA损伤测试(9.3 - 158倍);在膜联蛋白V死细胞试剂盒凋亡测试中(1.1 - 12.7倍)高于半胱天冬酶3/7试剂盒(1 - 6.5倍)。细胞周期分析表明,24小时后,MC3T3 - E1细胞处于G0/G1期的细胞数量(约70%)高于NE - 4C细胞(约40%)。目标物质的抑制效力顺序明确为POE - 15>农达经典>>草甘膦IPA盐。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b947/9301602/a32a6e61fc51/ga1.jpg

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