炎症诱导早产小鼠脑组织长链非编码RNA的筛选及验证研究

陈茹娟; 奚莎; 王凡; 肖谧; 林晓洁; 刘俐

doi:10.7499/j.issn.1008-8830.2016.05.012

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中国当代儿科杂志 ›› 2016, Vol. 18 ›› Issue (5) : 435-439. DOI: 10.7499/j.issn.1008-8830.2016.05.012

论著·实验研究

炎症诱导早产小鼠脑组织长链非编码RNA的筛选及验证研究

陈茹娟, 奚莎, 王凡, 肖谧, 林晓洁, 刘俐

作者信息 +

Screening and validation of long non-coding RNAs in brain tissue of inflammationinduced preterm mice

CHEN Ru-Juan, XI Sha, WANG Fan, XIAO Mi, LIN Xiao-Jie, LIU Li

Author information +

文章历史 +

摘要

目的探讨长链非编码RNA(lncRNA)与炎症诱导早产小鼠脑损伤的关系,为脑损伤的防治提供依据。方法孕鼠腹腔注射LPS建立炎症诱导早产小鼠脑损伤模型(早产组),正常孕鼠分娩足月小鼠作为对照(足月组)。应用lncRNA芯片技术筛选与早产小鼠脑损伤相关的lncRNA,并应用Real-timePCR技术对lncRNA进行验证。结果比较早产组与足月组,显著差异表达的lncRNA有1978条(P<0.05),包括上调lncRNA786条,下调lncRNA1192条,其中差异表达1.5倍及以上的lncRNA共有29条。对差异表达倍数最高的前10条lncRNA行进一步分析,发现这些lncRNA可能参与转录、信号转导、凋亡、细胞周期、炎症反应等生物学过程,并与G蛋白偶联受体信号通路、神经肽信号通路有关。对其中2条lncRNA在早产组和足月组脑组织中的表达进行Real-timePCR验证,发现与芯片结果一致。结论炎症诱导早产小鼠脑组织中的lncRNA表达谱发生了明显变化,G蛋白偶联受体信号通路可能参与早产脑损伤的发生机制。

Abstract

Objective To investigate the association between long non-coding RNAs (lncRNAs) and brain injury in inflammation-induced preterm mice, and to provide a reference for the prevention and treatment of brain injury. Methods An intraperitoneal injection of lipopolysaccharide in pregnant mice was performed to establish a model of inflammation-induced preterm mice with brain injury (preterm group). The full-term mice delivered by normal pregnant mice were used as controls (full-term group). The lncRNA chip assay was used to screen out the lncRNAs associated with brain injury in preterm mice. Quantitative real-time PCR was used to validate the lncRNAs identified by the above method. Results The preterm and full-term groups showed significant differences in the expression of 1 978 lncRNAs (P<0.05), consisting of 786 up-regulated lncRNAs and 1 192 down-regulated lncRNAs, and 29 lncRNAs were 1.5 or more times differentially expressed between the two groups. A further analysis was performed for the 10 most differentially expressed lncRNAs, and the results showed that these lncRNAs were involved in the biological processes including transcription, signal transduction, apoptosis, cell cycle, and inflammatory response, as well as G proteincoupled receptor signaling pathway and neuropeptide signaling pathway. Real-time PCR was performed to validate the expression of two lncRNAs in brain tissue in the preterm and full-term groups, and the results were consistent with those of the chip assay. Conclusions The expression profiles of lncRNAs in brain tissue change significantly in inflammation-induced preterm mice, and the G protein-coupled receptor signaling pathway may be involved in the pathogenesis of preterm brain injury.

导出引用

陈茹娟, 奚莎, 王凡, 肖谧, 林晓洁, 刘俐. 炎症诱导早产小鼠脑组织长链非编码RNA的筛选及验证研究[J]. 中国当代儿科杂志. 2016, 18(5): 435-439 https://doi.org/10.7499/j.issn.1008-8830.2016.05.012

CHEN Ru-Juan, XI Sha, WANG Fan, XIAO Mi, LIN Xiao-Jie, LIU Li. Screening and validation of long non-coding RNAs in brain tissue of inflammationinduced preterm mice[J]. Chinese Journal of Contemporary Pediatrics. 2016, 18(5): 435-439 https://doi.org/10.7499/j.issn.1008-8830.2016.05.012

参考文献

[1] Yuan TM, Sun Y, Zhan CY, et al. Intrauterine infection/ inflammation and perinatal brain damage: role of glial cells and Toll-like receptor signaling[J]. J Neuroimmunol, 2010, 229(1-2): 16-25.
[2] Salmaso N, Jablonska B, Scafidi J, et al. Neurobiology of premature brain injury[J]. Nat Neurosci, 2014, 17(3): 341-346.
[3] Fancy SP, Harrington EP, Baranzini SE, et al. Parallel states of pathological Wnt signaling in neonatal brain injury and colon cancer[J]. Nat Neurosci, 2014, 17(4): 506-512.
[4] Nagano T, Fraser P. No-nonsense functions for long noncoding RNAs[J]. Cell, 2011, 145(2): 178-181.
[5] Ponting CP, Oliver PL, Reik W. Evolution and functions of long noncoding RNAs[J]. Cell, 2009, 136(4): 629-641.
[6] Mercer TR, Dinger ME, Mattick JS. Long non-coding RNAs: insights into functions[J]. Nat Rev Genet, 2009, 10(3): 155-159.
[7] Wilusz JE, Sunwoo H, Spector DL. Long noncoding RNAs: functional surprises from the RNA world[J]. Genes Dev, 2009, 23(13): 1494-1504.
[8] Ng SY, Lin L, Soh BS, et al. Long noncoding RNAs in development and disease of the central nervous system[J]. Trends Genet, 2013, 29(8): 461-468.
[9] Qureshi IA, Mehler MF. Emerging roles of non-coding RNAs in brain evolution, development, plasticity and disease[J]. Nat Rev Neurosci, 2012, 13(8): 528-541.
[10] Zheng B, Liu H, Wang R, et al. Expression signatures of long non-coding RNAs in early brain injury following experimental subarachnoid hemorrhage[J]. Mol Med Rep, 2015, 12(1): 967-973.
[11] Mehta SL, Kim T, Vemuganti R. Long noncoding RNA FosDT promotes ischemic brain injury by interacting with RESTassociated chromatin-modifying proteins[J]. J Neurosci, 2015, 35(50): 16443-16449.
[12] Wang F, Xiao M, Lin XJ, et al. Expression of heme oxygenase 1 and leukemia inhibitory factor in maternal plasma and placental tissue in a lipopolysaccharide induced late-pregnancy preterm birth mouse model[J]. J Reprod Med, 2016, 61(1-2): 39-46.
[13] Faghihi MA, Modarresi F, Khalil AM, et al. Expression of a noncoding RNA is elevated in Alzheimer's disease and drives rapid feed-forward regulation of beta-secretase[J]. Nat Med, 2008, 14(7): 723-730.
[14] 杨峰, 易凡, 曹慧青, 等. 长链非编码RNA 研究进展[J]. 遗传, 2014, 36(5): 456-468.
[15] Hangauer MJ, Vaughn IW, McManus MT. Pervasive transcription of the human genome produces thousands of previously unidentified long intergenic noncoding RNAs[J]. PLoS Genet, 2013, 9(6): e1003569.
[16] Giera S, Deng Y, Luo R, et al. The adhesion G proteincoupled receptor GPR56 is a cell-autonomous regulator of oligodendrocyte development[J]. Nat Commun, 2015, 6: 6121.
[17] Bae BI, Tietjen I, Atabay KD, et al. Evolutionarily dynamic alternative splicing of GPR56 regulates regional cerebral cortical patterning[J]. Science, 2014, 343(6172): 764-768.
[18] Singer K, Luo R, Jeong SJ, et al. GPR56 and the developing cerebral cortex: cells, matrix, and neuronal migration[J]. Mol Neurobiol, 2013, 47(1): 186-196.