Department of Biology, School of Natural Sciences, University of Patras, GR-26500 Rio, Patras, Greece.
Department of Sustainable Agriculture, School of Agricultural Sciences, University of Patras, GR-30131 Agrinio, Greece.
Sci Total Environ. 2024 Dec 1;954:176484. doi: 10.1016/j.scitotenv.2024.176484. Epub 2024 Sep 24.
The present study investigated the oxidative and cytogenotoxic potential of Tetraethylene glycol dimethyl ether (known as Tetraglyme) on healthy human peripheral blood lymphocytes, widely used as an in vitro model for assessing the human health risk posed by different chemical compounds. In a first step, Nuclear Magnetic Resonance (H NMR) spectroscopy, and Ultra-High Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) were employed to estimate Tetraglyme's stability under a wide range of pH values (4-12), and thus to identify potential by-products. Thereafter, isolated lymphocytes were treated with different concentrations of Tetraglyme (0.02-20 mg L) for assessing its oxidative (using the DCFH-DA staining), and cytogenotoxic potential (using the trypan blue exclusion test for estimating cell viability, Comet assay, as well as the cytokinesis-block micronucleus assay, with or without the addition of S9 metabolic activation system). According to the results, Tetraglyme remains stable at pH 4, but two additional derivatives (i.e. 1-[2-(2-ethoxyethoxy)ethoxy]-2-methoxyethane [CHO] and 1-ethoxy-2-(2-ethoxyethoxy)ethane (Diethylene glycol diethyl ether) [CHO]) were found in traces, under alkaline conditions (pH ≥7). Moreover, although Tetraglyme (and/or its derivatives) showed negligible alterations of cell viability (>92 %) in all cases, the pronounced ROS formation, DNA damage, cell proliferation arrest, and MN frequencies in challenged cells are indicative of its oxidative and cytogenotoxic potential. The significant alterations of Cytokinesis-Block Proliferation Index (CBPI) and Micronucleus (MN) frequencies in S9 challenged cells give further evidence for the potential involvement of Tetraglyme's metabolites in the observed cytogenotoxic mode of action. Although not conclusive, the present findings give rise to further research, utilizing different cell types and biological models, for elucidating Tetraglyme's toxic mode of action, as well as its environmental and human risk.
本研究旨在探讨四乙二醇二甲醚（又称四甘醇二甲醚）对健康人外周血淋巴细胞的氧化和细胞遗传毒性，该细胞常用于评估不同化学化合物对人类健康风险的体外模型。在第一步中，采用核磁共振（NMR）波谱和超高效液相色谱-质谱联用（UHPLC-MS）技术，研究了四甘醇二甲醚在广泛 pH 值（4-12）范围内的稳定性，并由此确定了潜在的副产物。随后，用不同浓度的四甘醇二甲醚（0.02-20 mg L）处理分离的淋巴细胞，以评估其氧化（使用 DCFH-DA 染色）和细胞遗传毒性（使用台盼蓝排斥试验评估细胞活力、彗星试验以及有丝分裂阻断微核试验，同时或不添加 S9 代谢激活系统）。结果表明，四甘醇二甲醚在 pH 值为 4 时保持稳定，但在碱性条件下（pH≥7），还发现了两种额外的衍生物（即 1-[2-(2-乙氧基乙氧基）乙氧基]-2-甲氧基乙烷[CHO]和 1-乙氧基-2-(2-乙氧基乙氧基）乙烷（二乙二醇二乙醚）[CHO]）。此外，尽管四甘醇二甲醚（和/或其衍生物）在所有情况下对细胞活力的改变可忽略不计（>92%），但在受到挑战的细胞中，ROS 形成、DNA 损伤、细胞增殖停滞和微核频率的明显增加表明其具有氧化和细胞遗传毒性。在添加 S9 的细胞中，细胞有丝分裂阻断增殖指数（CBPI）和微核（MN）频率的显著改变进一步证明了四甘醇二甲醚代谢物可能参与了观察到的细胞遗传毒性作用模式。尽管这并不具有结论性，但这些发现引发了进一步的研究，利用不同的细胞类型和生物学模型，阐明四甘醇二甲醚的毒性作用模式及其环境和人类风险。
