PC12细胞转染人GPx-1基因后的抗氧化损伤作用  

Anti-oxidative damage effect on PC12 cells transfected with human glutathione peroxidase 1 gene

作　　者：马琳[1] 王辉[2] 王淑荣[1] 陈蓉[1] 郑俩燕[1] 

机构地区：[1]海南医学院附属医院神经内科,海南海口570102 [2]西安交通大学医学院第二附属医院老年神经科,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2013年第4期460-464,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省2008科技计划项目(No.2008k15-05)~~

摘　　要：目的探讨GPx-1高表达在H2O2介导的PC12细胞损伤时的保护作用及分子机制。方法将PC12细胞转染含人GPx-1基因的plncx质粒及空载体plncx质粒,经持续筛选建立起稳定表达人GPx-1基因和空质粒plncx的PC12细胞系。对PC12、GPx-1-PC12、plncx-PC12 3组细胞,分别给予H2O2和亚硒酸钠+H2O2干预方式,GPx活性检测试剂盒检测各组细胞GPx活性;MTT法检测细胞的存活情况;流式细胞仪检测细胞凋亡发生的情况;免疫细胞化学法观察NF-κBp65的表达情况。结果 GPx-1-PC12组细胞GPx活性较对照组明显增高;3组细胞分别给予2种干预后,PC12与plncx-PC12组细胞存活率比较,差异无统计学意义(P>0.05),GPx-1-PC12较PC12、plncx-PC12组细胞存活率明显增高(P<0.01);GPx-1-PC12较PC12、plncx-PC12细胞早期和晚期凋亡率明显下降,存在统计学差异(P<0.01);GPx-1-PC12较PC12细胞NF-κBp65表达明显减少(P<0.01)。结论 GPx-1基因高表达对H2O2所致的PC12细胞氧化损伤有很好的保护作用;GPx-1可抑制PC12细胞氧化损伤时凋亡的发生及NF-κB的表达。Objective To study the protective effect and molecular mechanism of glutathione peroxidase 1（GPx-1） high expression on PC12 cell damage mediated by hydrogen peroxide（H2O2）.Methods PC12 cells were transfected with plncx plasmid containing human GPx-1 gene and plncx plasmid,and a PC12 cell line was established that could stably express human GPx-1 gene and plncx plasmid through continuous screening.Two means of intervention,H2O2 and sodium selenite ＋ H2O2,were given to PC12,GPx-1-PC12 and plncx-PC12 cells,respectively.GPx activity of cells in each group was detected with GPx activity detection kit;the survival and apoptosis of cells were detected by MTT method and flow cytometer,respectively.The expression of NF-κBp65 was detected by immunohistochemistry technique.Results GPx activity of GPx-PC12 group cells was obviously higher than that of controls.Intervention of H2O2 and sodium selenite ＋ H2O2 did not significantly affect the cell survival rate in PC12 and plncx-PC12 groups（P0.05）.The cell survival rate was higher in GPx-1-PC12 group than in PC12 and plncx-PC12 groups（P0.01）;the early and late apoptosis rates were obviously lower in GPx-1-PC12 group than in PC12 and plncx-PC12 groups（P0.01）.The positive rate of NF-κBp65 protein decreased significantly in GPx-1-PC12 group compared with that in PC12 group（P0.01）.Conclusion The overexpression of GPx-1 gene has a great protective effect on the oxidative damage of PC12 cell mediated by H2O2.GPx-1 can inhibit the occurrence of PC12 cell apoptosis and NF-κB expression during oxidative damage.

关 键 词：PC12细胞 谷胱甘肽过氧化物酶1(GPx-1) 核因子-kappaB(NF-κB) 过氧化氢(H2O2) 细胞凋亡 

分 类 号：R741.02[医药卫生—神经病学与精神病学]