高糖状态下人视网膜色素上皮细胞氧化应激损伤情况以及上皮-间质转化情况评估

Assessment of the oxidative stress damage of human retinal pigment epithelium and epithelial-mesenchymal transition under high glucose state

目的:研究高糖环境对人视网膜色素上皮(RPE)细胞氧化应激损伤情况以及上皮-间质转化的影响。方法:培养人RPE细胞,并分为低糖对照组、高渗对照组、高糖干预组,分别用葡萄糖浓度为5.5mmol/L的DMEM培养基、渗透浓度为60.0mmol/L且葡萄糖浓度为5.5 mmol/L的DMEM培养基、葡萄糖浓度为60.0mmol/L的DMEM培养基处理,24 h后测定细胞中氧化应激分子、细胞凋亡分子以及间质细胞标志分子的含量。结果:高糖干预组细胞中ROS、MDA、3-NT、Nrf2、ARE、Caspase-3、Bax、JNK、c-JUN、α-SMA、Vimentin、N-cadherin的含量均显著高于低糖对照组和高渗对照组,GST、HO-1的含量均显著低于低糖对照组和高渗对照组;低糖对照组和高渗对照组细胞中ROS、MDA、3-NT、Nrf2、ARE、Caspase-3、Bax、JNK、c-JUN、α-SMA、Vimentin、N-cadherin、GST、HO-1的含量无显著性差异。结论:高糖环境能够通过增强RPE细胞的氧化应激反应来启动细胞的凋亡过程以及上皮间质转化过程。

Objective：To study the effect of high glucose environment on oxidative stress injury of human retinal pigment epithelium（RPE）as well as pithelial-mesenchymal transition.Methods：Human RPE cells were cultivated and divided into low glucose control group,hyperosmosis control group and high glucose intervention group that were treated with DMEM medium with glucose concentration 5.5 mmol/L,DMEM medium with osmotic concentration 60.0 mmol/L and glucose concentration5.5 mmol/L as well as the DMEM medium with glucose concentration60.0mmol/L respectively,and after 24 h,the levels of oxidative stress molecules,cell apoptosis molecules and mesenchymal cell marker molecules in cells were determined.Results：ROS,MDA,3-NT,Nrf2,ARE,Caspase-3,Bax,JNK,c-JUN,α-SMA,Vimentin and N-cadherin levels in high glucose intervention group were significantly higher than those of low glucose control group and hyperosmosis control group while GST and HO-1 levels were significantly lower than those of low glucose control group and hyperosmosis control group;ROS,MDA,3-NT,Nrf2,ARE,Caspase-3,Bax,JNK,c-JUN,α-SMA,Vimentin,N-cadherin,GST and HO-1levels werenot significantly different between low glucose control group and hyperosmosis control group.Conclusions：High glucose environment can enhance the oxidative stress response of RPE cells to start the cell apoptosis and epithelial-mesenchymal transition process.