摘要
目的探讨化学性低氧模拟剂氯化钻(CoCl_2)对小鼠视网膜神经节细胞(RGC)中核因子E2相关因子2(Nrf2)及其下游调控的抗氧化酶超氧化物歧化酶(SOD)和过氧化氢酶(CAT)表达的影响。方法以小鼠视网膜神经节细胞株RGC-5为研究对象,四甲基偶氮唑蓝法(MTT)检测细胞生长活性筛选最适CoCl_2作用浓度;最适浓度的CoCl_2作用RGC-5细胞2、6、12、24h,Western Blot检测细胞内总的及细胞核内Nrf2蛋白的表达;Real-time RT-PCR检测SOD和CAT的mRNA表达。结果最适CoCl_2浓度为200μmol/L,CoCl_2作用2hRGC-5细胞内和细胞核内Nrf2蛋白表达水平开始上调,6h达高峰,12h下降,24h恢复正常。而CoCl_2作用2h,SOD和CATmRNA在RGC-5细胞表达水平开始上升,12h达高峰,24h开始回落但仍高于正常组。结论化学性低氧模拟剂CoCl_2能够诱导细胞内Nrf2蛋白表达,促其转位进入细胞核,上调其介导的抗氧化酶SOD和CAT基因的转录。
Objective To investigate the effect of chemical hypoxia-mimetic agent（cobalt chloride,CoCl_2） on the expressions of Nrf2 and its downstream antioxidant enzymes（SOD,CAT） in mouse retinal ganglion cells（RGC-5）.Methods MTT was used to detect the growth activity of RGC-5 for screening the best suitable concentration of CoCl_2 to establish the hypoxia model.The total level and nuclear level of Nrf2 protein in RGC-5 cells were evaluated by Western blot,and the expressions of SOD mRNA and CAT mRNA were measured by Real-time RT-PCR.Results The optimal concentration of CoCl_2 was 200μmol/L,the total level and nuclear level of Nrf2 protein in RGC-5 cells were increased 2h after administration of CoCl_2,peaked at 6 h,declined 12 h,and turned to normal level 24 h.The expression levels of SOD mRNA and CAT mRNA in RGC-5 cells were increased 2h after administration of CoCl_2,peaked 12 h？ and began to decline 24 h but still higher than normal cells.Conclusion CoCl_2 induced the expression and translocation of Nrf2 into the nucleus of RGC-5 cells,and activated the transcription of the regulated downstream antioxidase SOD and CAT in RGC-5 cells.
出处
《解剖科学进展》
2016年第1期26-28,共3页
Progress of Anatomical Sciences
基金
辽宁省自然科学基金(2014020184)
