硫化氢对H_2O_2诱导的视网膜神经节细胞氧化损伤的保护作用  

Protective effects of H_2S against H_2O_2-induced oxidative injury in RGC

作　　者：罗鑫[1] 刘太祥[1] 任荟璇 罗艳[1] 

机构地区：[1]遵义医学院附属医院贵州省眼科医院,贵州省遵义市563003

出　　处：《眼科新进展》2017年第6期515-518,共4页Recent Advances in Ophthalmology

基　　金：贵州省科学技术基金资助(编号:20147588);遵义医学院博士启动基金(编号:201307)~~

摘　　要：目的探讨硫化氢对H_2O_2诱导的视网膜神经节细胞(retinal ganglion cell,RGC)-5氧化损伤的保护作用及可能机制。方法将RGC-5分为4组,RGC-5组为正常对照组;氉RGC-5+H_2O_2组:RGC-5在500μmol·L^(-1)H_2O_2中培养24 h诱导氧化损伤;RGC-5+NaH S+H_2O_2组:RGC-5置于50μmol·L^(-1)NaH S中30 min后在500μmol·L^(-1)H_2O_2中培养24 h;RGC-5+NaH S组:RGC-5置于50μmol·L^(-1)NaH S中30 min。Western blot检测线粒体内、外细胞色素C和视神经萎缩蛋白1(optic atrophy 1,OPA1)表达;用荧光探针JC^(-1)检测线粒体膜电位,用透射电镜观察线粒体形态。结果与RGC-5组相比,RGC-5+H_2O_2组RGC-5细胞质内细胞色素C表达增加,而线粒体内的OPA1表达减少(均为P<0.05)。RGC-5组和RGC-5+NaH S+H_2O_2组线粒体内、外细胞色素C的表达差异均无统计学意义(均为P>0.05)。与RGC-5组相比,RGC-5+NaH S组细胞质内细胞色素C表达减少,而线粒体内的细胞色素C表达增加(均为P<0.05)。与RGC-5组相比,RGC-5+H_2O_2组RGC-5细胞质内OPA1表达显著增加,而线粒体内的OPA1表达减少(均为P<0.05)。RGC-5+NaH S+H_2O_2组和RGC-5+NaH S组RGC-5线粒体内、外OPA1的表达与RGC-5组相似,差异均无统计学意义(均为P>0.05)。RGC-5+H_2O_2组线粒体膜电位与其他3组比较明显下降,其余3组间线粒体膜电位比较,差异无统计学意义(P>0.05)。RGC-5+H_2O_2组线粒体肿胀呈球状,而其余3组线粒体肿胀不明显。结论硫化氢可能通过阻止线粒体释放OPA1来减轻H_2O_2导致的RGC-5氧化损伤。Objective To investigate the protective effects and possible mecha- nism of hydrogen sulfide （H2S） against oxidative stress injury induced by hydrogen per- oxide （ H2O2 ） in retinal ganglion cell-5 （ RGC-5 ）. Methods RGC-5 cells were divided into four groups： RGC-5 group （ normal control group）, RGC-5 ＋ H2O2 （ RGC-5 were cul- tured in 500 μmol L-1 H2O2 for 24 hours） group,RGC-5 ＋ NariS （ RGC-5 were cultured in 50 μmol L-1 NariS for 30 minutes） ＋ H2O2 （ RGC-5 were cultured in 500 μmol L-1 H2O2 for 24 hours） group, and RGC-5 ＋ NariS （ RGC-5 were cultured in 50 μmol L-1 NariS for 30 minutes） group. Western blots were applied to measure the expression of cytochrome c （Cyt. c） and optic atrophy 1 （OPAl）. The fluorescent dye JC-1 assay was chosen to detect the mitochondrial membrane potential （Am）, Furthermore ,transmis- sion electron microscope was used to observe the morphology of mitochondria. Re. suits Compared with RGC-5 group, the expression of Cyt. c in RGC-5 ＋ H2O2 group decreased in mitochondria, and increased in cytoplasm （ all P 〈 0.05 ）, but there was no statistical difference between RGC-5 group and RGC-5 ＋ NariS ＋ H2O2 group （ all P 〉 0. 05 ）. Compared with RGC-5 group, the expression of Cyt. c in RGC-5 ＋ NariS group in- creased in mitochondria, and decreased in cytoplasm （ all P 〈 0. 05 ）. Compared with RGC-5 group, the expression of OPAl in RGC-5 ＋ H2O2 group decreased in mitochondri- a, and increased in cytoplasm （ all P 〈 0.05 ）. In RGC-5 ＋ NaHS ＋ H： O2 group and RGC- 5 ＋ NariS group, the expression of OPAl within and outside the mitochondria had no significant difference compared with RGC-5 group （ all P 〉 0.05 ）. Compared with other three groups, the mitochondrial membrane potential in RGC-5 ＋ H2O2 group obviously decreased, but there was no statistical difference among other three groups （P 〉 0.05 ）. The mitochondria were globular swelled in RGC-5 group, but in other three groups, th

关 键 词：氧化应激 视网膜神经节细胞 硫化氢 视神经萎缩蛋白1 

分 类 号：R774[医药卫生—眼科]