[C_(8)mim]Cl对HepG2细胞凋亡和内质网应激通路的影响

Effects of[C_(8)mim]Cl on apoptosis and endoplasmic reticulum stress pathway in HepG2 cells

为研究离子液体氯化1-辛基-3-甲基咪唑([C8mim]Cl)是否通过内质网应激(ERS)通路诱导细胞凋亡,在MTT法检测细胞活力的基础上,用0,50,100,200μmol/L[C8mim]Cl处理HepG2细胞24h后,采用流式细胞仪检测细胞凋亡,western blot检测ERS通路相关蛋白表达.结果显示:[C8mim]Cl处理后HepG2细胞凋亡呈浓度依赖性增高.ERS相关蛋白葡萄糖调节蛋白78(GRP78)、磷酸化RNA依赖的蛋白激酶样内质网激酶(p-PERK)、磷酸化真核起始因子2α(p-eIF2α)、磷酸化肌醇需求酶-1(p-IRE1)、激活转录因子4(ATF4)和ATF6显著上调.[C8mim]Cl还显著诱导了C/EBP同源蛋白(CHOP)和半胱氨酸天冬氨酸蛋白酶4(caspase 4)蛋白表达,促进了caspase 9和caspase 3活性升高.因此,[C8mim]Cl可通过ERS通路诱导HepG2细胞凋亡.

In order to explore whether ionic liquid 1-octyl-3-methylimidazolium chloride([C8mim]Cl)can induce apoptosis through endoplasmic reticulum stress(ERS)pathway,HepG2 cells were treated with 0,50,100,200μmol/L[C8mim]Cl for 24h based on the MTT cell viability assay.The apoptosis was detected by flow cytometry and western blot was used to assay the expressions of ERS related proteins.The results showed that[C8mim]Cl increased the apoptosis in HepG2 cells with a concentration-dependent manner.[C8mim]Cl significantly increased the expressions of ERS related proteins,including glucose regulated protein 78(GRP78),phosphorylated protein kinase RNA-like endoplasmic reticulum kinase(p-PERK),phosphorylated eukaryotic initiation factor 2α(p-eIF2α),phosphorylated inositol-requiring enzyme 1(p-IRE1),activating transcription factor 4(ATF4)and ATF6.[C8mim]Cl also significantly induced the expressions of C/EBP homologous protein(CHOP)and caspase 4 protein,lead to the increase of caspase 9 and caspase 3 activities.Therefore,[C8mim]Cl could induce apoptosis in HepG2 cells through ERS pathway.