离子液体氯化1-辛基-3-甲基咪唑对EMT6细胞的毒性及其机理研究

为评估咪唑类离子液体的生物毒性,研究了氯化1-辛基-3-甲基咪唑(C8mim] C1])对EMT6细胞的毒性作用和可能的机制.不同浓度(0.06、0.25、1 mmol·L-)的C8 mim]Cl]对EMT6细胞染毒12h后,采用MTT方法检测细胞活力,二乙酸荧光素(FDA)方法检测细胞膜通透性的变化,Rhodamine 123染色方法检测线粒体膜电位的变化,ELISA方法检测了Caspase-3的活性,并测定了细胞内活性氧(ROS)的含量.结果表明,经Csmim] Cl]染毒12h后,EMT6细胞活力下降,并呈剂量依赖关系.当Csmim] Cl]浓度高于0.25 mmol·L-1时,细胞活力与对照相比,差异显著.研究还发现,Csmim]Cl]染毒增加了EMT6细胞膜通透性,降低了线粒体膜电位并诱导产生过量的活性氧,增强了Caspase-3活性.实验结果表明,C8mim] Cl]染毒造成了EMT6细胞膜通透性的改变、活性氧的过量产生和凋亡分子表达的增强,这可能是Csmim] Cl]导致细胞凋亡和活力下降的主要原因.

Cytotoxicity and possible mechanism of 1-octyl-3-methylimidazolium chloride on EMT6 cells

Cytotoxicity and possible mechanism of 1-octyl-3-methylimidazolium chloride (C₈mim]Cl]) on EMT6 cells were investigated in the present study. The cytotoxicity was evaluated by MTT assay, plasma membrane permeability was detected by FDA-staining, the change of mitochondrial membrane potential was determined by Rhodamine 123, the activity of Caspase-3 was determined by ELISA, and the intracellular ROS levels were also detected in EMT6 cells after exposure for 12 h at 0.06, 0.25, 1 mmol·L^-1 of C₈mim]Cl], respectively. The results showed that C₈mim]Cl] inhibited EMT6 cell growth and decreased their viabilities in a dose-dependent manner after exposure for 12 h. Compared to the control group, the difference of cell viability was significant when the concentration of C₈mim]Cl] was more than 0.25 mmol·L^-1. We also found that -exposure increased the plasma membrane permeability, decreased mitochondrial membrane potential, induced overproduction of intracellular ROS, and enhanced the activities of Caspase-3. These results suggest that C₈mim]Cl] may induce EMT6 cell apoptosis mediated by permeability transition and mitochondrial depolarization and triggered by excessive ROS and enhanced-expression of apoptotic molecules.