基于高通量测序分析并验证非动脉炎性前部缺血性视神经病变患者外周血miRNAs表达谱差异  

Analysis and verification of peripheral blood miRNAs expression profile difference in non-arteritis anterior ischemic opticc neuropathy patients based on high-throughput sequencing

作　　者：王群[1,2] 王欣玲 WANG Qun;WANG Xin-Ling(Department of ophthalmology,the Fourth Affiliated Hospital of China Medical University,Shenyang 110005,Liaoning,China)

机构地区：[1]中国医科大学附属第四医院眼科,辽宁沈阳110005 [2]辽宁省晶状体学重点实验室

出　　处：《中国老年学杂志》2025年第6期1341-1346,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金(No.81570838)。

摘　　要：目的基于非动脉炎性前部缺血性视神经病变(NAION)患者外周血的高通量测序数据,运用生物信息学方法分析miRNAs的表达谱改变并进行验证。方法收集2021年5月至2022年5月就诊于中国医科大学附属第四医院眼科的8例NAION患者及6名健康对照组的肘静脉血液样本,提取RNA后进行高通量测序分析,筛选差异表达的基因后利用生物信息学方法进行靶基因、基因本体论(GO)分类功能注释分析、京都基因与基因组百科全书(KEGG)通路富集分析,构建ceRNA网络,实时定量反转录聚合酶链反应(qRT-PCR)技术验证差异基因表达水平。结果共筛选出33个差异表达的miRNA,其中18个呈上调趋势,15个呈下调趋势。差异表达miRNAs在GO富集分析主要富集在细胞器生物黏附、生物调节、细胞发育、生长免疫系统、突触部分结合、转录因子活性、催化活性、转运蛋白活性等过程;KEGG通路富集分析探究到多种代谢过程,包括加压素调节、醛固酮合成、肾上腺素能信号转导系统、上皮细胞、鞘磷脂、破骨细胞分化及丝裂原活化蛋白激酶(MAPK)信号传导等。qRT-PCR验证结果显示,miR-6859-5p、miR-216a-5p在NAION患者外周血中表达上调,miR-3607-5p、miR-4690-3p表达下调。结论NAION患者较健康对照者外周血中基因表达谱发生显著变化,差异表达的基因可能参与了NAION发生发展过程中多个信号通路。ObjectiveBased on the high-throughput sequencing data of peripheral blood of patients with non-arteritic anterior ischemic optic neuropathy(NAION)to analyze and verify the changes of miRNAs expression profile by bioinformatics it.Methods Elbow vein blood samples were collected from 8 NAION patients and 6 healthy controls from the Department of Ophthalmology,the Fourth Affiliated Hospital of China Medical University from May 2021 to May 2022.RNA was extracted and analyzed by high throughput sequencing.After screening differentially expressed genes,bioinformatics methods were used to analyze target genes,gene ontology(CO)classification function annotation analysis,Kyoto gene and genome Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis,to construct ceRNA network,and real-time fluorescent quantitative reverse transcription polymerase chain reaction(qRT-PCR)technology was used to verify the expression level of differential genes.Results A total of 33 differentially expressed miRNA were screened,of which 18 were up-regulated and 15 were down-regulated.In GO enrichment analysis,differentially expressed miRNAs was mainly enriched in organelle bioadhesion,biological regulation,cell development,growth and immune system,synaptic partial binding,transcription factor activity,catalytic activity,transporter activity and so on.A variety of metabolic processes were explored by KEGG pathway enrichment analysis,including vasopressin regulation,aldosterone synthesis,adrenergic signal transduction system,epithelial signal transduction,sphingomyelin signal transduction,osteoclast differentiation and mitogen activated protein kinase(MAPK)signal transduction.The results of qRT-PCR verification showed that expressions of miR-6859-5p and miR-216a-5p were upregulated,expressions of miR-3607-5p and miR-4690-3p were down-regulated in peripheral blood of patients with NAION.Conclusions The gene expression profile in peripheral blood of patients with NAION is significantly dfferent from that of control group,and the dif

关 键 词：非动脉炎性前部缺血性视神经病变 高通量测序 MIRNA 

分 类 号：R774.6[医药卫生—眼科]