大鼠心力衰竭细胞microRNA表达谱及生物信息学分析  

Screening of specific microRNAs and bioinformatics analysis on cardiomyocytes in rat with heart failure

作　　者：朱海娟 何淑芳[2] 金世云[2] 胡军[2] 张野[2] 

机构地区：[1]安徽省妇幼保健院麻醉科,合肥230061 [2]安徽医科大学第二附属医院麻醉科,合肥230601

出　　处：《中华急诊医学杂志》2016年第4期439-443,共5页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金青年基金（81200171）;安徽省科技厅年度重点项目（1301043031）

摘　　要：目的：观察大鼠心力衰竭细胞中microRNA （ miRNA）表达变化，利用生物信息学分析技术探索差异miRNA靶基因的功能。方法18只成年雄性SD大鼠，体质量200-220g，随机数字表法随机分为两组：正常对照组（ CON组）和心力衰竭组（ HF组）。 HF组制备阿霉素致心力衰竭模型， CON组注射等量生理盐水。提取大鼠心脏， Largendorff逆行灌注法分离心室肌细胞，提取总RNA行miRNA表达谱检测，筛选两组差异表达的miRNA，荧光定量RT-PCR 技术验证结果， Targetscan、 miRanda软件预测差异表达miRNA的靶基因，并对靶基因行生物信息学分析。结果芯片检测结果显示，与CON组相比， HF组共有37个miRNA表达发生显著改变，其中22个miRNA上调，15个miRNA下调（均P＜0.01， FDR＜0.05）。荧光定量RT-PCR检测miR-133b-5p （t ＝14.56， P＜0.1）、 miR-6216（t＝9.32， P＜0.1）、 let-7e-5p （ t＝13.92， P＜0.1）表达水平，变化趋势与芯片结果一致。生物信息学分析显示，差异表达miRNA调控的靶基因显著富集于31个基因组（均P＜0.01， FDR＜0.05）和12条信号通路（均P＜0.05， FDR＜0.05），其中泛素蛋白酶体系统、 MAPK信号通路、 Toll样受体信号通路富集程度较高。结论阿霉素诱导的心力衰竭模型大鼠心肌细胞中miRNA表达谱发生显著改变，这些差异表达miRNA可能通过调控靶基因功能参与心力衰竭的病理生理过程。Objective To investigate microRNAs （ miRNAs） expression profiling of cardiomyocytes in rats with heart failure, and predict miRNAs-regulated target genes and their functions.Methods Total of 18 male SD rats weighing 200-220 g were randomly divided into 2 groups：the control group （ CON） and the heart failure group （HF）.The rats in HF group were injected by adriamycin via tail vein to induce heart failure, meanwhile in CON group, rats were received an equal volume of 0.9% sodium chloride intravenously.The cardiomyocytes isolated from the rat hearts in two groups and cultured overnight.After that, total RNA was extracted and then subjected to miRNA microarray to screen differentially expressed miRNAs.The reults of microarray were further verified by quantitative real-time PCR （ qRT-PCR ） .The target genes regulated by differentially expressed miRNAs were predicted by the software of Targetscan and miRanda.Bioinformatics analysis was performed to predict the miRNAs-regulated target genes and analyze the enriched gene ontology （ GO） and signaling pathway （ KEGG Pathway） .Results The results of miRNA microarray showed that a total of 37 miRNAs were differentially expressed in HF group as compared to CON group, among which 22 miRNAs were up-regulated and 15 miRNAs were down-regulated （P〈0.01, FDR〈0.05）.The expression of miR-133b-5p （t=14.56, P〈0.01）, miR-6216 （t=9.32, P〈0.01） and let-7e-5p （t=13.92, P〈0.01） which were detected by qRT-PCR exhibited the similar tendency of up or down regulation to those shown in microarray results.Bioinformatics analysis indicated that miRNAs-regulated target genes were significantly enriched in 31 GOs （P〈0.01, FDR〈0.05） and 12 signal pathways （P〈0.05, FDR〈0.05）, among which ubiquitin-proteasome system, MAPK signaling pathway and Toll like siganling pathway exhibited a higher enrichment. Conclusion MiRNA expression profile on cardiomyocytes in rat with adriamycin-induced heart failure was significantly changed.These diffe

关 键 词：心力衰竭 心肌细胞 MICRORNA表达谱 生物信息学分析 荧光定量RT—PCR 靶基因 基因组 KEGG通路 

分 类 号：R541.6[医药卫生—心血管疾病]