高氧暴露对早产新生大鼠肺组织转录因子NF-E2相关因子2和Keap1表达的影响  

Effects of hyperoxia on expression of nuclear factor-erythroid 2-related factor 2 and Keap1 in prema-ture newborn rats＇ lung

作　　者：蔡成[1,3] 吴军华 陈黎丽[1] 王明欢[1] 赵焕虎[2] 

机构地区：[1]宁波大学医学院附属宁波市妇女儿童医院NICU,315012 [2]中央民族大学中国少数民族传统医学研究所,北京100081 [3]上海交通大学附属儿童医院新生儿科,200062

出　　处：《中国小儿急救医学》2015年第10期685-688,692,共5页Chinese Pediatric Emergency Medicine

基　　金：浙江省自然科学基金资助项目（Y2110273）

摘　　要：目的探讨高氧暴露对早产新生大鼠肺组织中转录因子NF-E2相关因子2（ nuclearfactor-erythroid 2-related factor 2, Nrf2）和胞浆蛋白伴侣分子Keap1表达的影响。方法早产新生SD大鼠生后1d完全随机设计方法分为两组：高氧组与空气组。高氧组持续暴露于常压氧舱中，氧质量浓度〉85％；空气组置于同一室常压空气中。两组分别于高浓度氧或空气暴露后1…4710、14d提取肺组织标本。采用石蜡包埋切片行HE染色，观察肺组织的病理学变化。采取RT-PCR方法测定Nrf2和KeaplmRNA水平的动态表达。Westernblot检测Nrf2蛋白水平的表达。结果（1）与空气组相比，高氧组4、7d早产鼠肺组织Nrf2mRNA表达显著增强（4d：0．314±0．064VS．0．521±0．086，7d：0．440±0．121VS．0．658±0．076），10d出现下降趋势，14d明显减弱（P〈0．05）。（2）与空气组相比，高氧组1、4d肺组织中KeaplmRNA表达均显著增强（1d：0．352±0．052VS．0．547±0．075，4d：0．363±0．074VS．0．658±0．076）（P〈0．05），但7d后呈下降趋势，10、14d明显减弱（P〈0．05）。（3）与空气组相比，高氧暴露1、4d，Nrf2蛋白水平表达稍增强（P〉0．05）；7d后Nrf2蛋白呈下降趋势，其表达较空气组减弱，但7d两者相比差异无统计学意义（P〉0．05），而10、14d显著减弱（10d：1．325±0．464VS．0．755±0．348，14d：1．662±0．474VS．0．867±0．115）（P〈0．05）。结论氧化暴发诱导肺组织中Nrf2与Keapl的异常表达，调节机体氧化应激水平，参与高氧肺损伤发病过程。高氧暴露可能通过上调Nrf2活性，提高机体抗氧化，在减轻高氧肺损伤过程中发挥一定抗氧化作用。Objective To explore the expression of nuclear factor-erythroid 2-related factor 2 （Nrf2） and the molecular chaperone of cytoplasmic Keapl in premature newborn rats exposed to hyperoxia. Methods Completely randomized design method was performed, one-day old preterm SD rats were randomly divided into two groups：hyperoxia group and air group. The preterm SD rats in hyperoxia group were continuously exposed to oxygen （oxygen 〉 0. 85 ）and air group in room air. After 1,4,7,10,14 days of exposure, the pre- term SD rats of two groups were sacrificed, whole lung of these rats were isolated, the lung histological chan- ges were observed by HE staining. Total lung RNA was extracted, Nrf2 and Keapl mRNA were detected by RT-PCR. Western-blot was used to detect the changes of Nrf2 protein expression. Results （ 1 ） Compaired with air group ,the expression of Nrf2 in lung tissue of hyperoxia group significantly increased after 4,7 days of exposure（4 d：0. 314±0. 064 vs. 0. 521 ±0. 086,7 d：0. 440±0. 121 vs. 0. 65± 0. 076） （P 〈0. 05） ,the general tendency decreased after 10 days,but the expression of Nrf2 became significantly weak after 14 days of exposure in hyperoxia group （P 〈 0.05 ）. （2） The expression of Keapl mRNA in hyperoxia group signifi- cantly increased in 1,4 days（ 1 d：0. 352± 0. 052 vs. 0. 547 ± 0. 075,4 d：0. 363 ± 0. 074 vs. 0. 658 ±0. 076 ） （P 〈 0. 05 ）, the general tendency decreased after 7 days of exposure, but the expression of Keapl mRNA in hyperoxia group became significantly weaker than its expression after 10,14 days of air group （ P 〈 0.05 ）. （3） In comparison with air group ,Nrf2 protein expression in hyperoxia group increased after 1,4 days of ex- posure with no significant difference（P 〉0. 05） ,but had a tendency of decreasing after 7 days. On day 10, 14,its expression in hyperoxia group became significantly weak compared with that of air group （ 10 d：1. 325 ±0.464 vs. 0.755 ±0.348,14 d：1.662 ±0.474 vs. 0.867±0. 115�

关 键 词：高浓度氧 肺损伤 转录因子NF-E2相关因子2 早产 KEAP1 

分 类 号：R722.6[医药卫生—儿科]