侯天芳¹,
廖纪萍¹,
马元元²,
张成¹,
王广发¹
1. 北京大学第一医院呼吸和危重症医学科, 北京 100034;
2. 北京大学第一医院实验动物中心, 北京 100034

作者简介: 侯天芳(1988-),女,博士研究生,研究方向为大气污染与气道疾病,E-mail:tf_hou@bjmu.edu.cn.

基金项目: 国家自然科学基金面上项目(No.81370106)；北京市自然科学基金重点项目(No.7161013)；首都卫生发展专项研究重点攻关项目(No.2016-1-4071)；北京大学临床研究项目(No.PUCRP201303)；国家重点研发计划(No.2017YFC1309500)


中图分类号: X171.5

Microarray-based Analysis of Differentially Expressed MicroRNAs in Mouse Lung Tissue Induced by Particulate Matter

Hou Tianfang¹,
Liao Jiping¹,
Ma Yuanyuan²,
Zhang Cheng¹,
Wang Guangfa¹
1. Department of Respiratory and Critical Care Medicine, Peking University First Hospital, Beijing 100034, China;
2. Department of Laboratory Animal Center, Peking University First Hospital, Beijing 100034, China

CLC number: X171.5

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摘要:microRNAs作为临床疾病早期诊断的新型生物标记物越来越受到重视，为了进一步探究其在大气污染暴露后引起疾病的分子染毒机制。本研究通过建立大气污染小鼠染毒模型，利用Agilent芯片筛查小鼠肺组织中microRNAs差异表达谱，并通过实时荧光定量PCR方法验证芯片结果，使用TargetScan，PITA，microRNAorg数据对差异miRNA进行靶基因预测，靶基因富集的基因功能(GO)和信号通路(KEGG)分析。结果显示，大气细颗粒物暴露2周后小鼠肺组织microRNAs有显著差异表达谱，高剂量暴露组与对照组比较有4个miRNAs上调，低剂量暴露组与对照组比较有2个miRNAs上调，高剂量组与低剂量组比较，有4个miRNAs上调(标准为fold change值>= 2.0且P值<= 0.05)，挑选差异明显的miRNAs进行qRT-PCR验证，miR-139-5p、miR-691及miR-340-3p变化趋势与芯片一致，生物信息学结果显示，差异表达的miRNAs所调控的靶基因明显富集于34个GO通路(包括RNA转录酶II启动子的转录，RNA拼接，DNA模板，蛋白质结合和核酸结合)和32个KEGG通路(主要集中轴突导向通路和癌症通路)。综上所述，大气细颗粒物暴露染毒可诱导小鼠肺组织中miR-139-5p、miR-691及miR-340-3p明显上调，且生物信息学分析提示中枢神经系统发育及癌症通路可能作为PM_2.5暴露相关差异表达miRNAs调控靶基因介导的主要致病通路。
关键词: 大气污染/
PM_2.5/
microRNA/
生物信息学

Abstract:MicroRNAs can be regarded as new biomarkers for the early diagnosis of clinical diseases, which has been paid more and more attentions. In order to investigate the pathogenic mechanism of air pollution, BALB/c mice were treated with noninvasive tracheal instillation of PM_2.5 suspension to construct experimental animal model. Differentially expressed microRNAs in mouse lung tissues were screened by Agilent chip, and the results were verified by real-time quantitative PCR. Target genes prediction of differential miRNAs was performed using TargetScan, PITA, and microRNAorg. Then, target gene enrichment, gene function (GO) and signal pathway (KEGG) analysis were performed, and differentially expressed miRNAs were detected after exposed to particulate matter for 2 weeks. 4 miRNAs were up-regulated in high dose exposure group compared to control group, 2 miRNAs were up-regulated in low dose exposure group compared to control group, and 4 miRNAs were up-regulated in high dose exposure group compared to low dose exposure group (standard value > = 2 and the P value < = 0.05). The expression of miR-139-5p, miR-691 and miR-340-3p detected by qRT-PCR exhibited similar patterns of up-regulation to those shown in microarray results. Bioinformatics analysis revealed that the target genes of miRNAs were significantly enriched in 34 GOs (including transcription from RNA polymerase II promoter, RNA splicing, DNA-templated, protein binding, RNA binding, DNA binding) and 32 signal pathways (including mainly axon guidance and pathways in cancer). In conclusion, the expression levels of miR-139-5p, miR-691 and miR-340-3p in mice lung tissues were up-regulated remarkably after exposure to particulate matter. Bioinformatics analysis suggested that central nervous system development and cancer pathway may be the major pathogenic pathway mediated by differentially expressed miRNAs related to target genes.
Key words:air pollution/
PM_2.5/
microRNA/
bioinformatics analysis.