高氧诱导miRNA差异性表达介导新生大鼠的肺损伤  

Hyperoxia-induced differential expression of microRNAs mediates lung injury in neonatal rats

作　　者：王丽娜 章容 康兰 雷小平 何红霞 董文斌 Wang Lina;Zhang Rong;Kang Lan;Lei Xiaoping;He Hongxia;Dong Wenbin(Department of Neonatology,the Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属医院新生儿科,四川省出生缺陷中心,四川省泸州市646000

出　　处：《中国组织工程研究》2023年第11期1692-1700,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81571480),项目名称:去乙酰化酶SIRT1的SDMO修饰与早产儿高氧肺损伤的临床实验分析,项目负责人:董文斌。

摘　　要：背景:支气管肺发育不良主要的病理特征是肺发育停滞和较轻微的组织结构损伤,胎儿肺发育需经历5个时期,到囊状期的胎儿生后才可能存活,因此,早产、高氧诱导支气管肺发育不良的发病机制研究重点关注了囊状期和肺泡期。目的:探讨2个发育阶段——囊状期和肺泡期中起调控作用的差异miRNA在高氧相关的支气管肺发育不良中的作用。方法:将新生SD大鼠随机分为4组:0 d组(n=10)出生后即取肺组织;4,7,14 d组均设置2个亚组,高氧组(n=10)出生后进入持续高氧环境中(氧浓度85%-90%),分别于4,7,14 d后取肺组织,空气组(n=10)出生后进入空气环境中(吸入氧浓度21%),分别于4,7,14 d后取肺组织。采用miRNA-Seq技术获取肺组织miRNA并建库,Illumina平台测序,Deseq2(Version 1.10.1)、Venn图组间比较依次筛选DEmiRs,GO富集、PPI分析预测差异表达的miRNAs靶蛋白。结果与结论:①0 d组有633个miRNA,4,7,14 d组中的高氧亚组分别有609,614,584个miRNA,空气亚组分别有629,608,581个miRNA;②在出生0-4 d(囊状期)、4-7 d、4-14 d(肺泡期)时,高氧组分别获得130,3,114个差异表达的miRNAs,空气组分别获得106,0,111个差异表达的miRNAs,各时期两组均差异表达的miRNAs分别为91,0,79个;③在囊状期和肺泡期,高氧组新生鼠肺组织中miR-34a-5p、miR-21-5p显著表达,miR-34a-5p靶蛋白有198个,围绕PDGFRB/PDGFRA/NGFR;miR-21-5p靶蛋白302个,围绕MAPK1/STAT3;④结果显示,miR-34a-5p、miR-21-5p各自在囊状期和肺泡期受高氧诱导显著表达,分别调控靶蛋白PDGFRB/PDGFRA/NGFR、MAPK1/STAT3,介导阶段性肺发育过程的信号通路,参与支气管肺发育不良的发生发展。BACKGROUND:The main pathological features of bronchopulmonary dysplasia are stagnation of lung development and minor tissue damage.Fetal lung development needs to go through five stages,and a preterm infant cannot survive after birth until the cystic stage.Therefore,research on the pathogenesis of premature birth-and hyperoxia-induced bronchopulmonary dysplasia has focused on cystic and alveolar phases.OBJECTIVE:To investigate the regulatory role of differentially expressed microRNAs in hyperoxia-associated bronchopulmonary dysplasia during two developmental stagescystic and alveolar stages.METHODS:Newborn Sprague-Dawley rats were randomly into four groups:in 0 day group(n=10),rat lung tissue was to ken immediately after birth;4-,1-,14-day groups were further divided into two subgroupshyperoxia group(exposed to persistent hyperoxia with an oxygen concentration of 85%-90%,n=10)and air group(exposed to the air with FiO_(2):21%,n=10).Lung samples tissue were then to ken at postnatal days 4,7,14.MicroRNAs in lung tissue were acquired using miRNA-Seq technology to build a library,and sequenced on the Illumina platform.Differently expressed microRNAs between different groups were screened by Deseq2(Version 1.10.1)and Venn map.Gene ontology enrichment and protein-protein interaction analysis were performed to predict to rget proteins of differentially expressed microRNAs.RESULTS AND CONCLUSION:There were 633 microRNAs in the 0-day group,609,614,and 584 microRNAs in the hyperoxia group at postnatal days 4,7,14,respectively,and 629,608,and 581 in the corresponding air groups.The hyperoxia group obtained 130,3,and 114 differentially expressed microRNAs during postnatal days 0-4(cystic stage),4-7,and 4-14(alveolar stage),respectively.And the corresponding air group received 106,0,111 differentially expressed microRNAs respectively.There were 91,0,and 79 differentially expressed microRNAs in the two groups at different stages.During the cystic and alveolar phases,miR-34a-5p and miR-21-5p were significantly expressed in the l

关 键 词：支气管发育不良 miRNA高通量测序 miRNA-21-5p miR-34a-5p MAPK1/STAT3 PDGFRB/PDGFRA/NGFR 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R722