心房颤动伴脑卒中患者循环lncRNAs表达谱的初步研究

doi:10.13232/j.cnki.jnju.2022.01.017

南京大学学报(自然科学版) ›› 2022, Vol. 58 ›› Issue (1): 175–182.doi: 10.13232/j.cnki.jnju.2022.01.017

• • 上一篇

心房颤动伴脑卒中患者循环lncRNAs表达谱的初步研究

崔进进¹, 陈茜², 丛树一², 阮中宝¹^,²()

^1.南京中医药大学附属泰州人民医院，泰州，225300
^2.江苏省泰州市人民医院心血管内科，泰州，225300

收稿日期:2021-11-25 出版日期:2022-01-30 发布日期:2022-02-22
通讯作者: 阮中宝 E-mail:tzcardiac@163.com
作者简介:E⁃mail：tzcardiac@163.com
基金资助:
泰州市科技支撑计划

Genome⁃wide analysis of long non⁃coding RNA expression profile in patients with atrial fibrillation⁃related stroke

Jinjin Cui¹, Xi Chen², Shuyi Cong², Zhongbao Ruan¹^,²()

^1.Taizhou People's Hospital Affiliated to Nanjing University of Chinese Medicine，Taizhou, 225300, China
^2.Department of Cardiology, Jiangsu Taizhou People's Hospital, Taizhou, 225300，China

Received:2021-11-25 Online:2022-01-30 Published:2022-02-22
Contact: Zhongbao Ruan E-mail:tzcardiac@163.com

摘要/Abstract

摘要：

利用高通量测序筛选心房颤动（房颤）伴脑卒中患者外周血单核细胞中差异表达的lncRNAs，为从分子水平探讨房颤脑卒中发生发展机制提供实验依据，并寻找预测房颤脑卒中相关生物学标志物.应用高通量测序检测三例房颤伴脑卒中患者和三例房颤患者外周血单核细胞lncRNAs差异表达，进行基因本体论（Gene Ontology,GO）、代谢信号通路（Pathway）分析，建立lncRNAs?miRNAs共表达网络，并进行实时荧光定量PCR（qRT?PCR）验证.与对照组相比，房颤伴脑卒中患者差异表达301条lncRNA，其中上调143条，下调158条，上调和下调表达差异最大的是LINC01002，HCG18等基因.GO功能分析发现，差异表达基因参与疾病发生发展最主要的生物学过程包括结合、肽交联、高渗反应、内肽酶活性的调节、细胞体积的正向调节等.KEGG分析后发现，差异表达的lncRNA相关信号通路主要涉及免疫系统和炎症等相关.TCONS_00014860，TCONS_00020132，TCONS_00037601，TCONS_00003375，TCONS_00013755是lncRNAs?miRNAs共表达网络中结合节点最多的前五个lncRNAs，hsa?miR?619?5p是共表达网络中与lncRNAs作用最多的节点.qRT?PCR验证LINC01002，HCG18在两组人群中差异表达具有统计学意义.其与房颤相比，房颤伴脑卒中患者外周血存在差异表达lncRNA，与房颤发生脑卒中相关.XLOC_005045，XLOC_006945，XLOC_013107，XLOC_001139和XLOC_004677是共表达网络中结合节点最多的前五个lncRNAs，lncRNA?hsa?miR?619?5p可能在房颤伴脑卒中的发病机制中起到重要作用.同时，LINC01002，HCG18可以作为预测持续性房颤合并卒中的新型生物学标志物.

关键词: 心房颤动, 卒中, lncRNA, 高通量测序, 生物标志物

Abstract:

Using microarray to detect differentially expressed long non?coding RNAs (lncRNAs) in human Peripheral blood mononuclear cells (PBMC) from Atrial Fibrillation (AF)?related stroke patients and AF controls，we explore a theoretical basis for the mechanism of persistent AF?related stroke and search for novel biological markers. The peripheral blood monocytes of three patients with AF?related stroke and three AF subjects are collected for transcriptome sequencing (RNA?seq) to detect differently expressed lncRNAs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis are performed to identify the functions of differently expressed genes and related pathways. Then We verify the expression of LINC01002 and HCG18 through qRT?PCR. There are 301 differently expressed lncRNAs，including 143 up?regulated and 158 down?regulated expressions respectively. Besides，the most up?regulated and down?regulated expressions are LINC01002 and HCG18. GO analysis shows that differentially expressed genes enriched in differentially expressed transcripts in the biological process are mainly involved in copulation，peptide cross?linking，hyperosmotic response，regulation of endopeptidase activity，and positive regulation of cell volume process. KEGG enrichment pathway analysis shows that some of the enrichment pathways associate with differentially expressed lncRNAs include immune system and infectious diseases，etc. Network analysis uncover five key lncRNAs，which are XLOC_005045，XLOC_006945，XLOC_013107，XLOC_001139,and XLOC_004677. Hsa?miR?61?5p is the highest positive correlated miRNA in the networks. The results of qRT?PCR show that the expression of LINC01002 and HCG18 in the two groups are significantly different. There are differentially expressed lncRNAs in human monocytes from AF?related stroke patients. The lncRNAs?hsa?miR?619?5p?mediated network may play a critical role in the pathogenesis of AF?related stroke. LINC01002 and HCG18 can be novel biological markers for predicting AF?related stroke.

Key words: atrial fibrillation, stroke, lncRNA, RNA?seq, biological marker

中图分类号:

R541

崔进进, 陈茜, 丛树一, 阮中宝. 心房颤动伴脑卒中患者循环lncRNAs表达谱的初步研究[J]. 南京大学学报(自然科学版), 2022, 58(1): 175–182.

Jinjin Cui, Xi Chen, Shuyi Cong, Zhongbao Ruan. Genome⁃wide analysis of long non⁃coding RNA expression profile in patients with atrial fibrillation⁃related stroke[J]. Journal of Nanjing University(Natural Sciences), 2022, 58(1): 175–182.

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	房颤伴脑卒中组	对照组	P
性别(男性/%)	8（53%）	6（60%）	0.715
年龄(岁)	69.27±14.18	71±7.91	0.195
体重指数	23.07±3.57	24.15±3.467	0.901
吸烟指数(每年支数)	192.67±339.25	53.33±145.73	0.155
收缩压(mmHg)	139.47±18.20	132.87±22.92	0.383
舒张压(mmHg)	84.2±11.19	82.47±13.92	0.527
血糖(mmol·L^-1)	6.39±1.96	13.86±5.40	0.004
CHADS₂V_ASc评分	4.6±1.06	3.07±1.22	0.814
HASB?BLED评分	2.2±0.56	1.4±0.63	0.380
射血分数(%)	54.6±12.38	50.93±15.27	0.476
胆固醇(mmol·L^-1)	3.86±1.15	3.49±1.04	0.101
甘油三酯(mmol·L^-1)	1.52±1.28	1.26±0.58	0.700
高密度脂蛋白(mmol·L^-1)	1.06±0.25	1.05±0.36	0.554
低密度脂蛋白(mmol·L^-1)	2.34±0.88	2.12±0.718	0.331

差异基因	实验组表达	差异倍数	P
LINC01002	上调	144.69	0.000
LINC02289	上调	56.13	0.037
LINC01184	上调	50.51	0.000
FAM157C	上调	45.59	0.048
LOC105369519	上调	35.42	0.009
HCG18	下调	0.00	0.003
AC091057.3	下调	0.01	0.008
MUC20?OT1	下调	0.01	0.002
AL121985.1	下调	0.03	0.025
AC138965.2	下调	0.03	0.000

KEEG level 2 通路名称	该通路功能	P	富集分数
path:hsa04975	脂肪消化吸收	0.000	9.73
path:hsa00565	醚类脂代谢	0.000	7.93
path:hsa00600	鞘磷脂代谢	0.002	5.86
path:hsa00561	甘油脂代谢	0.003	5.04
path:hsa04666	Fc γ R介导的吞噬作用	0.009	3.72
path:hsa00564	甘油磷脂代谢	0.009	3.61
path:hsa05231	胆碱代谢	0.017	3.05
path:hsa04072	磷脂酶D信号传导途径	0.043	2.27

心房颤动伴脑卒中患者循环lncRNAs表达谱的初步研究

Genome⁃wide analysis of long non⁃coding RNA expression profile in patients with atrial fibrillation⁃related stroke

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