期刊导航

论文摘要

静脉桥模型大鼠在血管重塑过程中与失效有关的长链非编码RNA的筛选

The Screening for Long Non-coding RNAs Related to Vein Graft Failure in Rat External Jugular Vein-Carotid Bypass Models

作者:沈嘉渝, 卢晨, 张洪伟, 古君, 张煜, 胡佳

Author:SHEN Jia-yu, LU Chen, ZHANG Hong-wei, GU Jun, ZHANG Yu, HU Jia

收稿日期:2020-03-16          年卷(期)页码:2020,51(6):809-816

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:高通量测序, 长链非编码RNA, mRNA, 静脉桥失效

Key words:High-throughput sequencing, Long non-coding RNA, mRNA, Vein graft failure

基金项目:

中文摘要

目的 本研究旨在利用高通量测序技术分析长链非编码RNA(lncRNA)在大鼠自体静脉桥中的时间差异性表达,筛选出与静脉桥失效具有显著相关性的lncRNA。 方法 根据不同采样时间点将12只雄性SD大鼠随机分为4组 (分别为0 d、术后7 d、术后14 d、术后28 d组,每组3只)。其中以0 d组为对照组,其余3组为实验组(实验组大鼠制备颈外静脉-颈动脉旁路模型)。对静脉移植物样本进行高通量测序,并利用实时定量PCR(RT-qPCR)进行样本验证。通过识别在不同时间点差异表达的lncRNA及基因本体论(GO)富集分析和京都基因与基因组百科全书(KEGG)通路分析,对lncRNA-mRNA对进行功能预测,筛选出在静脉移植物管腔再狭窄过程中可能发挥潜在作用的lncRNA,并用RT-qPCR进行样本验证。 结果 高通量测序共获得了 2 572个静脉移植术后持续差异表达的lncRNA,同对照组(0 d)相比,分别在术后7、14、28 d差异表达程度前十位的lncRNA在文中列出。基于GO富集分析及KEGG通路分析结果,我们制定了以下筛选标准筛选出可能在冠脉动脉旁路移植术后移植物失效过程中发挥潜在关键作用的lncRNA-mRNA:①参与上述生物学过程或信号通路;② lncRNA可调节靶向mRNA转录;③既往已有报道靶向mRNA在血管病变过程中发挥作用;④lncRNA和mRNA均有显著性差异表达。最终筛选出包括MRAK083052-Nrp1在内的15对在静脉移植物管腔再狭窄过程中可能发挥潜在作用的lncRNA-mRNA。RT-qPCR结果与高通量测序结果一致。 结论 本研究发现大鼠静脉桥管腔再狭窄过程中lncRNA的表达变化情况与时间具有相关性。筛选并验证了15对在静脉移植物管腔再狭窄过程中可能发挥重要作用的lncRNA-mRNA。

英文摘要

ObjectiveThis study was designed to show the time-dependent changes in the expression profiles of long non-coding RNAs (lncRNAs) in rat vein grafts by using the high-throughput sequencing technique, and subsequently figure out lncRNAs related to vein graft failure.MethodsThe whole SD rats were randomly classified into four groups according to different sampling time points (0, 7, 14 and 28 days after surgery, respectively;n=3 per group). The day 0 group was set as a control, and the other three groups were set as experimental groups (the model of external jugular vein-carotid artery bypass was prepared in the experimental group). We identified the differentially expressed lncRNAs of the vein graft sample at different sampling time points with high-throughput sequencing, and verified these results using the technique of real-time quantitative PCR (RT-qPCR). Meanwhile, we used Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis to screen lncRNAs which may play roles in the restenosis process of vein grafts. The function of lncRNA-mRNA pairs was predicted. We subsequently used RT-qPCR to verify these lncRNAs.ResultsA total of 2 572 sustained differentially expressed lncRNAs were identified in our study. We showed the top ten differentially expressed lncRNAs at each post-operative time point. Through GO analysis and KEGG pathway analysis, we revealed the sustained differentially expressed genes which may be involved in VGF-related biological process, cellular component, molecular function and biological pathways. Finally, we screened 15 pairs of lncRNA-mRNA, includingMRAK083052-Nrp1, which may play roles in mediating the restenosis of vein graft. And the results of RT-qPCR were consistent with the results of the high-throughput sequencing.ConclusionThe present study investigated the time-dependent changes in the expression profiles of lncRNAs in vein grafts. We also screened 15 pairs of lncRNA-mRNA which may paly roles in vein graft failure.

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