Expressions of GM15886 and HIPK1 in lung tissues of nenatal mice with hyperoxia⁃induced lung injury
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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，及其可能的作用机制。方法：选取BPD模型小鼠肺组织lncRNA基因芯片中高表达的GM15886进行验证，应用IntaRNA、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，分别于生后第0、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和同源结构域相互作用蛋白激酶1（homeodomain-interacting protein kinase 1，HIPK1）表达水平；免疫组化检测HIPK1表达情况。结果：IntaRNA预测GM15886碱基序列能够和HIPK1第2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；GM15886可能靶向HIPK1参与BPD的发病机制。
Abstract:
Objective：To investigate the expression level of long non-coding RNA（lncRNA）GM15886 in lung tissues of bronchopulmonary dysplasia（BPD）induced by hyperoxia in neonatal mice and its mechanism in BPD. Methods：High expressed GM15886 was selected by previous lncRNA microarray. IntaRNA，Multi Experiment Matrix and Ensemble database were applied to predict GM15886 target genes. Sixty-four 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. Mice were sacrificed on day 0，day 3，day 5 and day 7，respectively，and the pathological changes of pulmonary tissues were analyzed via HE staining，the expression levels of GM15886 and homeodomain-interacting protein kinase 1（HIPK1）mRNA was detected by using QPCR，the expression of HIPK1 at different time points was detected by immunohistochemistry. Results：IntaRNA 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 day 3，day 5 and day 7，（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 day 3，day 5 and day 7 were 1.16±0.33，0.92±0.31，and 3.12±0.46，respectively. The expression level on day 7 was higher than that on day 0，day 3，day 5（all P < 0.05）. The expression of HIPK1 protein changed in the same way as mRNA. Conclusion：With the extension of hyperoxic exposure time，the expression of GM15886 in lung tissues increased gradually. GM15886 might participate in the pathogenesis of BPD by relugating HIPK1 expression.
