Fan Huimin, Niu Wei, He Mingjun, Kong Lingming, Zhong Aifang, Zhang Qiaoli, Yan Yan, Zhang Liyi
Cadre Ward, General Hospital of Chengdu Military Command, Chengdu, Sichuan 610083, P.R. China. Email:
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2015 Oct;32(5):641-6. doi: 10.3760/cma.j.issn.1003-9406.2015.05.007.
OBJECTIVE To identify differentially expressed microRNA (miRNA) in peripheral blood mononuclear cells (PBMCs) of anxiety patients and predict their target genes and function by bioinformatics analysis. METHODS The miRNA expression profiles were determined using an Affymetrix array. To validate the results, real-time quantitative polymerase chain reaction (qRT-PCR) analysis in a larger cohort was employed. The targets of the differentially expressed miRNAs were predicted by Target Scan, miRBD, and DIANA-microT-CDS, and the results were analyzed by gene ontology (GO) and KEGG pathway analysis using FunNet. RESULTS MicroRNA microarray chip analysis has identified 7 miRNAs were detected with significant changes in expression in PBMCs of anxiety patients. qRT-PCR analysis has confirmed that the expression levels of 5 miRNAs (has-miR-4484, has-miR-4505, has-miR-4674, has-miR-501-3p and has-miR-663) were up-regulated. Intersecting the genes by Target Scan, miRBD, and DIANA-microT-CDS has predicted 195 targets. GO analysis showed that biological processes regulated by the predicted target genes have included diverse terms. Some terms, e.g., nervous system development, nerve growth factor receptor signaling pathway, neuron migration, dendrite development, regulation of neuron projection development, midbrain development, regulation of excitatory postsynaptic membrane potential, gliogenesis, dendrite morphogenesis, etc. have direct relationship with the central nervous system and brain functions. Pathway analysis showed that a significant enrichment in several pathways related to neuronal brain functions such as glutamatergic synapse, axon guidance, calcium signaling pathway, MAPK signaling pathway, GnRH signaling pathway, Wnt signaling pathway, gap junction, long-term potentiation and VEGF signaling pathway, etc. Among the five microRNAs, has-miR-4484, has-miR-4505, has-miR-4674 and has-miR-501-3p may have more important regulatory functions. CONCLUSION Five miRNAs (has-miR-4484, has-miR-4505, has-miR-4674, has-miR-501-3p and has-miR-663) are up-regulated in PBMCs of anxiety patients and may be closely involved in the pathogenesis of anxiety disorder.
目的 鉴定焦虑症患者外周血单个核细胞（PBMCs）中差异表达的微小RNA（miRNA），并通过生物信息学分析预测其靶基因及功能。方法 使用Affymetrix芯片检测miRNA表达谱。为验证结果，在更大队列中采用实时定量聚合酶链反应（qRT-PCR）分析。通过Target Scan、miRBD和DIANA-microT-CDS预测差异表达miRNA的靶标，并使用FunNet通过基因本体论（GO）和KEGG通路分析对结果进行分析。结果 miRNA微阵列芯片分析已鉴定出7种miRNA在焦虑症患者的PBMCs中表达有显著变化。qRT-PCR分析证实5种miRNA（has-miR-4484、has-miR-4505、has-miR-4674、has-miR-501-3p和has-miR-663）的表达水平上调。通过Target Scan、miRBD和DIANA-microT-CDS对基因进行交叉分析预测出195个靶标。GO分析表明，预测的靶基因调控的生物学过程包含多种术语。一些术语，如神经系统发育、神经生长因子受体信号通路、神经元迁移、树突发育、神经元投射发育的调控、中脑发育、兴奋性突触后膜电位的调控、神经胶质生成、树突形态发生等，与中枢神经系统和脑功能有直接关系。通路分析表明，与神经元脑功能相关的几条通路有显著富集，如谷氨酸能突触、轴突导向、钙信号通路、MAPK信号通路、GnRH信号通路、Wnt信号通路、缝隙连接、长时程增强和VEGF信号通路等。在这5种微小RNA中，has-miR-4484、has-miR-4505、has-miR-4674和has-miR-501-3p可能具有更重要的调控功能。结论 5种miRNA（has-miR-4484、has-miR-4505、has-miR-4674、has-miR-501-3p和has-miR-663）在焦虑症患者的PBMCs中上调，可能密切参与焦虑症的发病机制。
