Toxicity Of Imidazoles Ionic Liquid [C₁₂mim]Cl/[C₁₆mim]Cl To HepG2,Hela And L929 Cells

Ionic liquids are low melting salt at room temperature.Since its many excellent physical and chemical properties,it is considered to be green solvent in the new century.In recent years,though for all kinds of application research progress of skyriver rapidly,however its harm to ecological environment and the organism has been neglected,research on toxicity of ionic liquids are rarely reported.In order to evaluate the toxicity of ionic liquids,we use HepG2,Hela and L929 cell line,studied the [C₁₂mim]Cl and [C₁₆mim]Cl on three kinds of cell toxicity,explore the possible toxic mechanism of cells.This study determined by MTT,single cell gel electrophoresis?comet assay?physiological toxicity tests?SOD?GSH?MDA?and flow cytometry,fluorescence quantitative methods,Then we detect the two kinds of ionic liquids on the toxic effects of HepG2?Hela and L929 cells.Results showed that the ionic liquids of [C₁₂mim]Cl and [C₁₆mim]Cl can inhibit the growth of HepG2,Hela and L929 cells,cause cell oxidative damage,and has genetic toxicity to the cells,induce apoptosis,influence the cell cycle,and apoptosis related gene p53,Bax,the quantity of the Bcl-2 expression.Determined by MTT experiment we found that toxicity of ionic liquids on the three kinds of cells have obvious dose effect relationship,and the toxic of [C₁₆mim]Cl is higher than [C₁₂mim] Cl.Hela and L929 cells are more sensitive to the drug.According to the determined by MTT experiment we concluded that the time of half inhibitory concentration,selected dosing interval for [C₁₂mim] Cl,we set up in 0.2,0.6,1.0,1.4 ?g/mL;for [C₁₆mim] Cl,we set up in 0.2,0.4,0.6,0.8 ?g/mL.Using single cell gel electrophoresis to check the ionic liquid [C₁₂mim]Cl(C₁₆H31ClN2)and [C₁₆mim]Cl?C20H39ClN2?on three kind of cells genetic damage.Results show that with the increase of drug concentration and the extension of time,the Tail DNA %,Tail Length,,Olive Tail moment have appeared different degree of rise.0.8 g/mL C₁₂ processing HepG2 cell,the Tail DNA % as high as 64.42%,tail length 441,Olive tail moment is 154.17.With the increase of dose and processing time,the nucleus has a comet-like tail,a nucleus head gradually narrowed,trailing degree increase gradually and become emission.Then based on the superoxide dismutase?SOD?,malondialdehyde?MDA?and reduced glutathione?GSH?content as an index,physiological toxicity test,to discuss whether the use of ionic liquids caused cell oxidative damage.Experimental group's MDA level is obvious rise,this may be due to the body through the enzyme system with the enzyme system to produce oxygen free radicals after the cells been damaged,and the rise of SOD and reduced GSH illustrate organism removal of oxygen free radical ability enhancement,also means that the cells undergo oxidative stress.Annexin V FITC/PI double dye apoptosis detection,the results showed that with the increase of ionic liquid concentration and time,cell necrosis and apoptosis rate increased.At the same time,we use the PI staining to detect changes in the cell cycle,found that with the increase of concentration and time,cells appeared the sub G1 phase,namely apoptosis peak.The two experiments have proved that the two kinds of ionic liquids can induced HepG2,Hela and L929 cell apoptosis.Using fluorescent real-time RT-PCR to detect the expression of the apoptosis related gene p53,Bax,Bcl–2,and the ratio of Bax/Bcl–2.Results showed that with the increase of ionic liquid concentration and time,the expression of p53 and Bax in three kinds of cell increased gradually,but the expression of Bcl-2 decreased gradually,the ratio of Bax/Bcl-2 also gradually rise.Showed that ionic liquids can through enhance the expression of p53 and Bax gene in HepG2,Hela and L929 cell,and reduce the expression of Bcl-2 inducing cell apoptosis.This study proved that two kinds of ionic liquids [C₁₂mim]Cl and [C₁₆mim]Cl on HepG2,Hela and L929 cell has the genetic toxicity,physiological toxicity and induction of apoptosis,it is not the true sense of green solvent.And the cell toxicity of [C₁₆mim]Cl is higher than [C₁₂mim]Cl.In addition,the drug resistance of HepG2 cell is higher than Hela and L929 cell.