体外新霉素损伤HEI-OC-1细胞的作用和机制研究

Effects of Neomycin on HEI-OC-1 Cells and Mechanisms-an in Vitro Study

目的目前尚不清楚HEI-OC-1细胞对耳毒性药物的反应和机制是否与耳蜗毛细胞相似。本研究以新霉素为耳毒性药物诱导HEI-OC-1细胞死亡,并探讨新霉素对HEI-OC-1细胞的损伤机制。方法HEI-OC-1细胞培养,加入新霉素诱导死亡,应用光镜、CCK-8检测HEI-OC-1细胞对新霉素的敏感性,应用流式细胞、免疫荧光、qRT-PCR检测HEI-OC-1细胞对新霉素的氧化应激机制。结果新霉素以浓度和时间依赖的方式诱导HEI-OC-1细胞死亡。氧化应激参与了HEI-OC-1细胞的凋亡和坏死。新霉素作用后,HEI-OC-1细胞的抗氧化-促氧化平衡被破坏,促进线粒体ROS积累,诱导MMP丢失,进而诱导HEI-OC-1细胞死亡。这种机制与新霉素在体内的损伤耳蜗毛细胞的机制相似。结论HEI-OC1细胞系可以提供一种细胞模型系统来研究新霉素等耳毒性药物对耳蜗毛细胞的损伤机制。

Objective Using neomycin as a toxic agent to induce HEI-OC-1 cells death to investigate underlying mechanisms of HEI-OC-1 cells injury by neomycin to help understand whether HEI-OC-1 cells reactions to ototoxic agents and mechanisms are similar to those in cochlear hair cells(HCs).Methods HEI-OC-1 cells were cultured and exposed to neomycin.The sensitivity of HEI-OC-1 cells to neomycin was detected by light microscope and CCK-8,and the oxidative stress was tested by flow cytometry,immunofluorescence and qRT-PCR.Results Neomycin induced death of HEI-OC-1 cells in a dose-and time-dependent manner.Besides,oxidative stress was involved in the apoptosis and necrosis of HEI-OC-1 cells.After neomycin treatment,antioxidant and pro-oxidative balance in HEI-OC-1 cells was destroyed,which promoted the accumulation of mitochondrial reactive oxygen species(ROS),induced the loss of mitochondrial transmembrane potentials(MMPs),and subsequently induced the death of HEI-OC-1 cells.These mechanisms were similar to in vivo neomycin injury in cochlear hair cells.Conclusion HEI-OC-1 cells may provide a cell model system for studying injury mechanisms of cochlear hair cells induced by neomycin and other ototoxic drugs.