早产儿氧暴露后外周血单个核细胞内活性氧簇产生与自噬变化的关系*

苏剑董文斌李清平康兰张莲玉卢佑英 翟雪松

(四川医科大学附属医院新生儿科, 四川 泸州 64600)

【摘要】目的观察早产儿氧暴露后外周血单个核细胞内活性氧簇产生与自噬的水平，探讨活性氧簇产生与自噬变化的关系。方法32周以下早产儿根据用氧浓度不同分为三组，FiO2<30%为低浓度吸氧组、FiO2 在30%～40%为中浓度吸氧组、FiO2>40%为高浓度吸氧组。各组氧疗48小后,与对照组中同期未吸氧的32周以下早产儿，经桡动脉采血3ml，分离外周血单个核细胞(PBMCs)及血清，采用激光共聚焦显微镜观察检测细胞内活性氧(ROS)的生成量，硫代巴比妥酸比色法检测血清丙二醛(MDA)含量，丹(磺)酰戊二胺(MDC)结合激光共聚焦显微镜观察自噬表达水平。结果早产儿氧暴露后，随着用氧浓度的升高，PBMCs内ROS。与其余三组相比，高浓度吸氧组中ROS，MDA含量和自噬体表达逐渐增高明显增加(P<0.05)，各组ROS水平与自噬表达水平呈正相关(P<0.01)。结论早产儿氧暴露后，高氧可诱导体内ROS增多，并在细胞内出现自噬现象。ROS可作为信号分子引起和上调细胞发生自噬反应。

【关键词】早产儿; 活性氧; 自噬

【中图分类号】R 722.6【文献标志码】A

基金项目：四川省教育厅科研课题(08ZA150)；四川省卫生厅科研课题(90191)

通讯作者：董文斌，教授，E-mail:DongWenbin2000@163.com

收稿日期：( 2015-01-09； 编辑： 张文秀)

The relationship between reactive oxygen species and autophagy within peripheral blood mononuclear cells in premature infants after oxygen exposed SU Jian, DONG Wenbin, LI Qingping,etal

(DepartmentofNewbornMedicine,TheAffiliatedHospitalofSichuanMedicalUniversity,Luzhou646000,Sichuan,China)

Abstract【】ObjectiveTo explore the relationship between reactive oxygen species and autophagy within peripheral blood mononuclear cells in premature infants after oxygen exposed. MethodsAccording to different oxygen concentrations, the preterm infants were divided into three groups including low concentration oxygen group (FiO2<30%), middle concentration oxygen group (FiO2 between30- 40%) and high concentration oxygen group (FiO2>40%). Premature infants less than 32 weeks without oxygen were as control group.After 48h, l blood were taken via radial artery and used to separate peripheral blood mononuclear cells(PBMCs)and serum. The intracellular reactive oxygen species (ROS) were detected by confocal laser scanning microscopy. The level of malondialdehyde (MDA)in serum were measured by thiobarbituric acid colorimetric. The expression of autophagy within peripheral blood mononuclear cells were observed by monodansylcadaverin combined with confocal laser scanningmicroscope.ResultsAfter premature infants exposed to oxygen, the oxygen, ROS and expression of autophagy and MDA were gradually increased. Compared that of the other three groups, the level of ROS, expression of autophagy and MDA in the high concentration oxygen group were increased significantly (P<0.05). There were positive correlations of autophagy and ROS in PBMCs among the groups(P<0.01). ConclusionHigh oxygen could induce in ROS increased in premature infants. The emergence of autophagy ROS could be used as a signal molecule induced upregulation of autophagy and cell reaction.

【Key words】Premature infants; ROS; Autophagy

近年来，随着医疗技术水平的发展，早产儿(特别是低出生体重儿)的存活率有了明显的提高。由于新生儿(特别是早产儿)呼吸系统疾病常常需要长时间吸入氧气，早产儿机体又缺乏抗氧化能力，则长时间、高浓度的氧疗易引起急慢性肺损伤的发生[1,2]。目前对于高浓度吸氧组诱导肺损伤的发病机制尚不清楚，但认为与氧化应激反应是有重要关系的[3]。……