College of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, China.
Institute of Bast Fiber Crops and Center of Southern Economic Crops, Chinese Academy of Agricultural Sciences, Changsha, 410205, China.
J Hazard Mater. 2018 Apr 15;348:1-9. doi: 10.1016/j.jhazmat.2018.01.028. Epub 2018 Jan 30.
Cytotoxicity studies are important tools for the assessment of the toxicity of ionic liquids (ILs). In the present study, the cytotoxicity of eleven ILs against Spodoptera frugiperda 9 (Sf-9) cell lines were evaluated via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. The effect on cellular morphology, ultrastructural morphology, and nuclear morphology induced by 1-ethyl-3-methylimidazolium bromide ([Cmim][Br]) was studied via inverted light microscopy observation, acridine orange staining, and transmission electron microscope (TEM) analysis, respectively. The effect on cell DNA fragmentation, cell apoptosis and cell cycle induced by [Cmim][Br] was also investigated via DNA agarose gel electrophoresis and flow cytometry analysis, respectively. The results showed that the cytotoxic effect of ILs on Sf-9 cells was related to the IL structures, concentrations, and length of exposure. The morphological features of apoptosis induced by [Cmim][Br] such as cell shrinkage and convolution, apoptotic bodies, pyknosis, and karyorrhesis were observed. All these phenomena confirmed that Sf-9 cells exposed to [Cmim][Br] died via apoptosis. This study complements the current knowledge about the cytotoxic properties of ILs on insect cells and highlights the mechanism by which ILs kill these cells. Furthermore, it provides a basis for further studies on the future applications of ILs as insecticides.
细胞毒性研究是评估离子液体（ILs）毒性的重要工具。在本研究中，通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法评估了十一 ILs 对 Spodoptera frugiperda 9（Sf-9）细胞系的细胞毒性。通过倒置相差显微镜观察、吖啶橙染色和透射电子显微镜（TEM）分析分别研究了 1-乙基-3-甲基咪唑溴化物([Cmim][Br])引起的细胞形态、超微结构形态和核形态的影响。还通过琼脂糖凝胶电泳和流式细胞术分析分别研究了[Cmim][Br]引起的细胞 DNA 片段化、细胞凋亡和细胞周期的影响。结果表明，ILs 对 Sf-9 细胞的细胞毒性效应与 IL 结构、浓度和暴露时间有关。观察到[Cmim][Br]诱导的凋亡的形态特征，如细胞皱缩和卷曲、凋亡小体、固缩和核碎裂。所有这些现象都证实 Sf-9 细胞通过凋亡死亡。这项研究补充了关于 ILs 对昆虫细胞的细胞毒性特性的现有知识，并强调了 ILs 杀死这些细胞的机制。此外，它为进一步研究 ILs 作为杀虫剂的未来应用提供了基础。
