高氧对胎鼠肺泡Ⅱ型上皮细胞NADH脱氢酶亚单位4、5表达的影响  

Effect of hyperoxia on expression of NADH-dehydrogenase subunit 4,5 in alveolar epithelial type Ⅱ cells from fetal SD rats

作　　者：黄光焰[1] 常立文[1] 汪鸿[1] 陈燕[1] 蔡成[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：《浙江医学》2010年第10期1478-1482,1485,共6页Zhejiang Medical Journal

摘　　要：目的 探讨高氧对胎鼠肺泡Ⅱ型上皮细胞（AEC Ⅱ）还原型烟酰胺腺嘌呤二核苷酸（NADH）脱氢酶亚单位4、5表达的影响.方法 原代培养Sprague-Dawley（SD）胎鼠AEC Ⅱ,待其生长至接近汇合状态时随机分为高氧组和正常对照组,高氧组持续暴露于常压高浓度氧中（氧浓度〉90%.CO2浓度5%）,正常对照组仍置于5%CO2培养箱中,两组均继续分别培养6、12、24h;采用免疫细胞化学染色SP法检测NADH脱氢酶亚单位4（DN4）蛋白的表达.采取半定量逆转录聚合酶链反应（RT-PCR）测定ND4、NADH脱氢酶亚单位5（ND5）mRNA的表达.结果 （1）SP法结果显示,与正常对照组比较,高氧组6h AEC Ⅱ ND4蛋白的表达增强,12h AEC Ⅱ ND4蛋白的表达减弱,但差异均无统计学意义（均P〉0.05）,24hAEC Ⅱ ND4蛋白的表达显著减弱（P〈0.05）;（2）RT-PCR检测结果显示,与正常对照组比较,高氧组6h AEC Ⅱ ND4、ND5 mRNA表达增强,差异均无统计学意义（均P〉0.05）.12、24h AECⅡ ND4、ND5 mRNA表达显著减弱（P〈0.05）.结论 持续高氧下调胎鼠AEC Ⅱ ND4、ND5 mRNA和AEC Ⅱ ND4蛋白的表达,这种改变可能参与高氧肺损伤的发病过程.Objective To investigate effect of hyperoxia on the expression of NADH-dehydrogenase subunit 4 （ND4）, NADH-dehydrogenase subunit 5 （ND5） in alveolar epithelial type Ⅱ cells （AEC Ⅱ ） from fetal SD rats. Methods Alveolar epithelial type Ⅱcells were obtained by primary culture from lungs of 19-d fetal SD rats. The cultured cells were randomly divided into con- trol and hyperoxia groups, cells in hyperoxia group were consistently exposed to hyperoxia （oxygen〉90%, carbon dioxide 5%）, while cells in control group were cultured in CO2 culture chamber （37~C, carbon dioxide 5%）. After culture for 6, 12 and 24 h, ND4 protein in cell slides were detected by immunocytochemistry, and ND4 and ND5 mRNA was detected by reverse transcription polymerase chain reaction （RT-PCR）. Results （1）Compared to control group, the expression of ND4 protein in hyperoxia group decreased significantly after exposure to hyperoxia for 24 h （P 〈 0.05）. （2）Compared to control group, the expression of ND4 and ND5 mRNA in hyperoxia group was decreased significantly after exposure to hyperoxia for 12 and 24 h （P 〈 0.05）. Conclusion Continuous exposure to hyperoxia down-regulates the expression of NADH-dehydrogenase subunit 4, 5 in alveolar epithelial type Ⅱ cells from fetal SD rats, which may be involved in the hyperoxia-induced lung injury.

关 键 词：高氧 胎鼠 肺泡Ⅱ型上皮细胞 NADH脱氢酶亚单位4 NADH脱氢酶亚单位5 

分 类 号：R383.34[医药卫生—医学寄生虫学]