小分子干扰RNA抑制高氧暴露下人肺腺癌A549细胞中的硫氧还蛋白-2对还原型烟酰胺腺嘌呤二核苷酸脱氢酶亚单位1、细胞色素C氧化酶I表达的影响  

Suppressed expression of thioredoxin-2 by small interference RNA in A549 cells exposed to hyperoxia reduced expression of nicotinamide-adenine dinucleotide dehydrogenase subunit 1 and cytochrome C oxidase I

作　　者：蔡成[1] 常立文[1] 李文斌[1] 陈燕[1] 单瑞艳[1] 刘伟[1] 潘睿[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科,武汉430030

出　　处：《中华围产医学杂志》2010年第2期128-133,共6页Chinese Journal of Perinatal Medicine

基　　金：国家自然科学基金（30672261、30770944）

摘　　要：目的探讨小分子干扰RNA（small interference RNA，siRNA）抑制高氧暴露下人肺腺癌A549细胞中硫氧还蛋白-2（thioredoxin-2，Trx-2）对还原型烟酰胺腺嘌呤二核苷酸（nicotinamide adenine dinucleotide，NADH）脱氢酶亚单位1（NADHdehydrogenase subunit1，NDl）及细胞色素c氧化酶I（eytochrome C oxidase I，COXI）表达的影响。方法体外传代培养A549细胞系，将Trx-2序列特异性的SiRNA转染A549细胞，随机分为无干扰空气组、无干扰高氧组、干扰后空气组与干扰后高氧组。分别于高氧或空气暴露12、24、48h提取A549细胞总RNA和蛋白。采用逆转录一聚合酶链反应技术测定Trx-2、ND1和COX I mRNA水平的表达，Western印迹技术检测Trx-2蛋白的表达。结果（1）序列特异性SiRNA可显著抑制Trx-2表达。（2）无干扰高氧组24hTrx-2mRNA水平为0．7799±0．1249，显著高于无干扰空气组（0．4424±0．1140）（P〈0．05）。干扰后高氧组24hTrx-2mRNA水平为0．2774±0．0174、48h为0．2587±0．0069，均显著低于干扰后空气组和无干扰高氧组（P〈0．05）。（3）无干扰高氧组24hND1 mRNA水平为0．6609±0．0368，显著低于无干扰空气组（0．8898±0．1049）（P〈0．05）。干扰后高氧组12hND1 mRNA水平为0．8848±0．0135，显著高于干扰后空气组（P〈0．05）。干扰后高氧组48hND1 mRNA水平为0．3808土0．0937，均显著低于干扰后空气组和无干扰高氧组（P〈0．05）。（4）无干扰高氧组12hCox 1 mRNA水平为1．7313±0．4331、24h为2．1929±0．6722、48h为2．0754±0．2584，均显著高于无干扰空气组（P〈0．05）。结论ND1、COXI参与高氧肺损伤发生发展过程，Trx-2对高氧肺损伤线粒体起重要的保护作用。Objective To explore the effects of expression of thioredoxin-2（Trx-2） suppressed by small interference RNA（SiRNA） in A549 cells exposed to hyperoxia on expression of nicotinamide adenine dinucleotide （NADH） dehydrogenase subunit 1 （ND1） and cytochrome C oxidase I （COX I ）. Methods A549 cells were gained by serial subcultivation in vitro and transfered with synthetic Trx-2 sequence-specific SiRNA and then were randomly divided into air group without interference, hyperoxia group without interference, air group after interference, and hyperoxia group after interference. After exposure to oxygen or room air for 12, 24 and 48 h, expressions of Trx-2, ND1 and COX I mRNA of these cells were detected by reverse transcription-polymerase chain reaction （RT-PCR）, and Trx-2 protein was detected by Western blot. Results （1）Sequence-specific SiRNA targeting Trx-2 could significantly down-regulate its expression in A549 cells. （2） Trx-2 mRNA levels in hyperoxia group without interference at 24 h was higher than those in air group without interference （0. 7799 ±0. 1249 vs 0. 4424±0. 1140, P〈0.05）. Trx-2 mRNA levels in hyperoxia group after interference at 24 h and 48 h were 0. 2774±0. 0174 and 0. 2587zk0. 0069,lower than those in air group after interference and hyperoxia group without interference （P 〈 0.05）. （3） ND1 mRNA levels in hyperoxia group without interference at 24 h was 0. 6609±0. 0368, lower than those in air group without interference （0. 8898±0. 1049）（P〈0. 05）. ND1 mRNA levels in hyperoxia group after interference at 12 h was 0. 8848_±0. 0135, higher than those in air group after interference（P〈0. 05）. ND1 mRNA levels in hyperoxia group after interference at 48 h was 0. 3808 ±0. 0937, lower than those in air group after interference and hyperoxia group without interference（P〈0. 05）. （4） COXI mRNA levels in hyperoxia group without inteference at 12, 24 and 48 h were 1. 7313±0. 4331, 2. 1929±0. 6722 and 2. 0754±0. 2584, higher than

关 键 词：RNA 小分子干扰 硫氧还蛋白质类 NADH脱氢酶 电子传递复合物IV 高氧症 肺疾病 

分 类 号：R722.6[医药卫生—儿科]