细胞色素b在早产新生大鼠高氧肺损伤肺组织中的表达和意义  

Dynamic changes of expression and significance of cytochrome b in premature rat lung during hyperoxia-induced lung injury

作　　者：蔡成[1] 常立文[1] 刘敬[1] 姜娜[1] 彭琼玲[1] 卢红艳[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,湖北武汉430030

出　　处：《临床儿科杂志》2007年第3期216-219,共4页Journal of Clinical Pediatrics

基　　金：国家自然科学基金资助项目(No.30471824)

摘　　要：目的探讨细胞色素b(cytochrome b,Cytb)与高氧肺损伤发生、发展的关系。方法早产新生SD(Sprague-Dawley)大鼠生后1d随机分为空气组、高氧组,高氧组持续暴露于常压氧舱中,氧浓度>85%;空气组置于同一室常压空气中。两组分别于高氧或空气暴露后1、4、7、10和14d时提取肺组织RNA,采用半定量逆转录聚合酶链反应(RT-PCR)测定Cytb mRNA表达;同时应用免疫组化方法检测肺组织切片中Cytb蛋白表达;应用Western-blot检测高氧肺损伤肺组织中Cytb蛋白水平的表达变化。结果与空气组相比,高氧暴露1d、4dCytb mRNA含量及其表达显著增强(P<0.05);7d后Cytb呈下降趋势,其表达较空气组减弱,但两者相比差异无统计学意义(P>0.05)。高氧暴露后,随日龄变化肺组织Cytb免化结果与Cytb mRNA表达相似;与空气组相比,高氧暴露1d、4d肺组织Cytb表达显著增强(P<0.05);7d后Cytb呈逐渐下降趋势,其表达较空气组减弱,但7、10d两组相比差异无统计学意义,14d极显著减弱(P<0.01)。Western-blot实验结果:与空气组相比,高氧暴露1d、4dCytb蛋白水平表达增强但两组相比差异无统计学意义(P>0.05);7d后Cytb呈下降趋势,其表达较空气组减弱,但7、10d两组相比差异无统计学意义(P>0.05),而14d显著减弱(P<0.05)。结论85%高浓度氧暴露诱导早产新生大鼠线粒体编码的Cytb异常表达,这种变化可能参与高氧肺损伤的发生。Objectives To investigate the relationship of cytochrome b and the development of hyperoxia-induced lung injury. Methods One-day-old preterm Sprague-Dawley （SD） rats were randomly divided into air group and hyperoxia group. Hyperoxia group was continuously exposed to oxygen （oxygen 〉 0.85） while the air group was just placed in room air. After 1, 4, 7, 10, 14 days of exposure, the rats were killed, whole lung RNA were isolated and cytochrome b mRNA were detected with reverse transcription polymerase chain reaction （RT-PCR） . Immunohistochemistry staining was used in lung sections to detect cytochrome b. Western-blot was used to detect the changes of protein expression during hyperoxia-induced lung injury. Results （ 1 ） In comparison with the air group, Cytochrome b mRNA expression was significantly higher （P 〈 0.05） after 1, 4 days of exposure to 85% oxygen; then decreased after 7 days, its expression became weak in comparison with air-breathing control rats with no significant difference （P 〉 0.05） . （2） Along with time varied, the results of Cytochrome b immunohistochemistry assay in lung section were in accordance with the Cytochrome b mRNA expression. With the expression of Cytochrome b significantly higher （P 〈 0.05） after 1, 4 days of exposure to 85% oxygen; and decreased after 7 days, its expression became weak in comparison with air-breathing control rats, with no significant difference （P 〉 0.05） . On day 14, Cytochrome b expression significantly decreased （P 〈 0.01） . （3） Results of Western-blot： in comparison with air group, Cytochrome b protein exression was increased after 1, 4 days of exposure with no significant difference （P 〉 0.05） but had a tendency of decreasing after 7 days, its expression became weak in comparison with air-breathing control rats with no significant difference after 7, 10 days of exposure （P 〉 0.05）. On day 14, its expression became significantly weak（P 〈 0.05） . Conclusions Exposure to 85% hyperoxia c

关 键 词：高氧 早产大鼠 细胞色素B 肺损伤 WESTERN-BLOT 

分 类 号：R722.6[医药卫生—儿科]