长链非编码RNA 1010001 N08 Rik在支气管肺发育不良形成中的表达规律及生物信息学分析

Expression of long non-coding RNA 1010001N08Rik in bronchopulmonary dysplasia and its bioinformatics analysis

目的 初步研究长链非编码RNA（long non-coding RNA,lncRNA） 1010001N08Rik在高氧诱导新生小鼠支气管肺发育不良（bronchopulmonarydysplasia,BPD）形成过程中的表达规律,并对其进行系统的生物信息学分析,预测其参与BPD发生发展的可能作用机制.方法 通过UCSC在线基因浏览工具获取1010001N08 Rik序列、所在基因组位置及其邻近基因相关信息;应用Ensemble数据库预测1010001N08Rik的靶基因.将新生C57BL/6J小鼠置于95％高氧环境下7d,制备高氧诱导新生小鼠BPD模型.采用逆转录聚合酶链反应检测1010001N08Rik、Gata 6 mRNA在BPD形成过程（第0、1、3、5、7天）中的变化情况,并通过t检验比较其表达差异.结果 1010001N08Rik在BPD形成过程第1、3、5、7天相对表达量分别为1.21±0.33、2.02±0.41、2.95±0.45、4.20±0.48,其表达逐渐增加,且第1、3、5、7天各组相近两个时点间差异均有统计学意义（P＜ 0.05）;Gata 6 mRNA第1、3、5、7天相对表达量分别为0.92±0.30、1.10±0.31、0.86±0.24、0.45±0.08,第7天低于第5天（P＜0.05）,其余相近两个时间点比较差异无统计学意义（P＞0.05）.1010001N08Rik在哺乳动物间具有较高的保守性,其所在染色体区域存在大量转录因子结合位点及表观遗传学线索,Gata 6为其可能的调控靶标.结论 1010001N08Rik在BPD形成过程中表达量逐渐增加,生物信息学分析及初步实验提示其可能通过下调Gata 6表达参与BPD形成过程.

Objective To explore the expression feature of long non-coding RNA （lncRNA） 1010001N08Rik in hyperoxia-induced bronchopulmonary dysplasia （BPD） and predict the mechanism that 1010001N08Rik might be involved in the occurrence and development of BPD by a series of bioinformatics analysis.Method The sequence,genomic position and structure characteristics of 1010001N08Rik were acquired from UCSC genome browser,and its target gene was predicted by Ensemble database.We successfully established the animal model of BPD by making newborn C57BL/6J mice exposed to 95% concentrations of ambient oxygen for seven days.The expression of 1010001N08Rik and Gata 6 were detected using real-time quantitative polymerase chain reaction （PCR）.Student＇s t test was used to compare their expression levels during the BPD process.Result The relative expression of 1010001N08Rik in BPD process at d1,d3,d5,d7 was 1.21 ± 0.33,2.02 ± 0.41,2.95 ± 0.45,4.20-± 0.48 respectively,and there were significant difference between adjacent time points （P 〈 0.05）.The relative expression of Gata 6 mRNA was 0.92 ±0.30,1.10 ± 0.31,0.86 ± 0.24,0.45-± 0.08 respectively,and there was significant difference between d5 and d7 （P 〈0.05）.1010001N08Rik had highly conserved property among different species.The chromosomal regions of 1010001N08Rik existed transcriptional factors binding locations and epigenetic regulation clues,and its possible candidate target gene was Gata 6.Conclusion The expression of 1010001N08Rik increased during the formation process of BPD.Bioinformatics analysis and preliminary experiment results suggested that 1010001N08Rik might participate in the process of BPD by down-regulating Gata 6 expression.