LncRNA GM15886在高氧诱导支气管肺发育不良新生小鼠肺组织的表达规律及生物信息学分析

[摘 要] 目的：探讨长链非编码RNA（long non-coding RNA，lncRNA）GM15886在高氧诱导新生小鼠支气管肺发育不良（bronchopulmonary dysplasia，BPD）肺组织中表达水平的变化,并运用生物信息学分析,预测其在BPD模型中的作用机制。方法：在已建立BPD模型小鼠肺组织lncRNA基因芯片中选取并验证GM15886，应用lncRNATargets、Multi Experiment Matrix和Ensemble数据库预测其靶基因；将64只新出生C57BL／6J小鼠随机分为高氧组和空气组，每组各32只；高氧组新生小鼠置于95％高氧环境下，空气组暴露于空气环境下，分别选取生后第0天、3、5、7天处死小鼠取肺组织，HE染色评价肺组织病理变化；采用qPCR检测GM15886、HIPK1 mRNA表达水平；免疫组化检测HIPK1表达情况。结果：lncRNATargets预测GM15886碱基序列能够和HIPK1第二个外显子碱基序列完全结合。qPCR示高氧组第3、5、7天GM15886表达量升高，分别为1.91±0.28、2.12±0.38、2.35±0.43，与第0天相比，差异均具有统计学意义（P均＜0.05）；HIPK1在第3、5、7天相对表达量分别为1.16±0.33、0.92±0.31、3.12±0.46，第7天表达量高于第0、3、5天（P均＜0.05）；HIPK1免疫组化表现与RNA表达的结果相对应。结论：随着高氧暴露时间的延长，肺组织GM15886表达量增加；生信分析和上述实验表明GM15886可能靶向HIPK1参与BPD的发病机制。

Expression of long non-coding RNA GM15886 in bronchopulmonary dysplasia and its bioinfomatics analysis

[Abstract] Objective: To investigate the expression level of long non-coding RNA GM15886 in lung tissues of bronchopulmonary dysplasia (BPD) induced by hyperoxia in neonatal mice and to conduct biological analysis to predict the regulation mechanism in BPD model. Methods: GM15886 was selected and verified by previous lncRNA microarray. lncRNATargets, Multi Experiment Matrix and Ensemble database were applied to predict GM15886 target genes.?64 newly born C57BL / 6J mice were randomly divided into the hyperoxic group and the air group, with 32 mice in each group. Newborn mice in the hyperoxia group were exposed to 95% oxygen, while those in the air group were exposed to air; The lung tissues of mice were sacrificed on 0 day, 3 day , 5 day and 7 day , respectively;the pathological changes of pulmonary tissues were excised for analysis via HE staining;QPCR was used to detect the expression levels of GM15886 and HIPK1 mRNA; The expression of HIPK1 at different time points was detected by immunohistochemistry. Results: LncRNATargets predicted the end of GM15886 sequence overlaps with the gene HIPK1.The expression of GM15886 in neonatal mice in the hyperoxia group increased gradually on 3d, 5d and 7d (1.91±0.28, 2.12±0.38 and 2.35±0.43, respectively), and the differences were statistically significant compared with day 0 (all P < 0.05).The relative expressions of HIPK1 on 3d, 5d and 7d were 1.16±0.33, 0.92±0.31 and 3.12±0.46, respectively. The expression level on 7d was higher than that on on 3d, 5d and 7d (all P < 0.05).The immunohistochemical expression of HIPK1 corresponded to the RNA expression. Conclusions: With the extension of hyperoxic exposure time, the expression of GM15886 in lung tissues increased gradually.Bioinformatics analysis and experiment results indicate that GM15886 might participate in the pathogenesis of BPD by relugating HIPK1 expression.