肝郁证青光眼大鼠视网膜miRNA表达特征及生物信息学分析  

miRNA expression profile and bioinformatics analysis of retina in liver depression glaucoma rats

作　　者：王丽媛 于天洋[2] 董霏雪 赵爽[1] 樊晓瑞 孙河[1] WANG Li-yuan;YU Tian-yang;DONG Fei-xue;ZHAO Shuang;FAN Xiao-rui;SUN He(Ophthalmology Department,First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院眼科,哈尔滨150040 [2]黑龙江中医药大学,哈尔滨150040

出　　处：《中华中医药杂志》2022年第1期501-507,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82004425,No.81674029,No.82074497);黑龙江中医药大学校科研基金资助项目(No.2019MS10,No.201510)。

摘　　要：目的:观察肝郁证青光眼大鼠视网膜微小核糖核酸(miRNA)的表达特征,探究肝郁证青光眼的发生机制。方法:将24只雄性SD大鼠随机分为空白组、青光眼组和肝郁证青光眼组,每组8只。青光眼组采用巩膜上静脉高渗盐水注射制作青光眼大鼠模型,肝郁证青光眼组采用慢性不可预知性温和应激刺激制作肝郁模型,造模成功后再进行慢性青光眼模型的制备。通过旷场实验检测大鼠肝郁证造模情况,采用Tono-pen avia笔式眼压计检测模型大鼠眼压,造模成功2周后,收集大鼠视网膜组织,采用HE染色观察视网膜组织形态学变化,提取视网膜总RNA,进行miRNA芯片检测,借助TargetScan、miRanda、miRBase数据库得到差异表达miRNA的靶基因;利用DAVID数据库和KEGG数据库进行富集分析;采用Cytoscape 3.7.1软件构建miRNA-mRNA网络。结果:本研究成功构建肝郁证青光眼大鼠模型,经miRNA芯片检测发现,与空白组比较,肝郁证青光眼组差异表达的miRNA共11个(上调2个,下调9个);与青光眼组比较,肝郁证青光眼组差异表达的miRNA共18个(上调1个,下调17个);与肝郁证青光眼特异相关的差异miRNA包括miR-133b-5p、miR-1843a-3p、miR-347等。富集分析显示,差异表达miRNA的靶基因主要富集在细胞黏附、血管生成、神经元凋亡等生物过程,集中在细胞质膜、高尔基体、突触等细胞组分中,涉及磷脂酰肌醇结合、GDP结合等分子功能及cGMP-PKG、cAMP等信号通路上。miRNA-mRNA网络显示,差异表达miRNA对应BDNF、mTOR等靶基因。结论:肝郁证青光眼模型大鼠视网膜组织中差异性表达的miRNA可能通过影响相关生物过程、分子功能和信号通路调控BDNF、mTOR等靶基因,影响肝郁证青光眼的发生发展。Objective:To observe the miRNA expression profile of retina in liver depression glaucoma rats,explore the mechanism of the occurrence of liver depression glaucoma.Methods:Twenty-four male SD rats were randomly divided into control group,glaucoma group and liver depression glaucoma group,8 rats in each group.Glaucoma group was established by superior scleral vein hypertonic saline injection.Liver depression glaucoma group was established liver depression by chronic unpredictable mild stress,after successful modeling,then establish glaucoma model as glaucoma group.The modeling of liver depression rats was detected by open-field experiment,and the intraocular pressure was detected by Tono-pen avia tonometer.After two weeks of successful modeling,the retinal tissues were taken for HE staining.Total retinal RNA was extracted for miRNA array detection,the target genes of differentially expressed miRNA were obtained by TargetScan,miRanda and miRBase database.Enrichment analysis of gene was carried out by DAVID database and KEGG database,respectively.Cytoscape 3.7.1 software was used to construct the network of miRNA-mRNA.Results:In this study,we successfully constructed liver depression glaucoma rat model.According to the miRNA array detection,compared with control group,11 miRNA were differentially expressed in the liver depression glaucoma group,of which 2 were up-regulated and 9 were down-regulated.Compared with the glaucoma group,18 miRNAs were differentially expressed in the liver depression glaucoma group,of which 1 was up-regulated and 17 were down-regulated.The different miRNAs specifically associated with liver depression glaucoma included miR-133 b-5 p,miR-1843 a-3 p and miR-347,etc..Enrichment analysis showed that the target genes of differentially expressed miRNA were mainly enriched in biological processes such as cell adhesion,angiogenesis and neuronal apoptosis;concentrated in cytoplasmic membrane,golgi body,synapse and other cell components;and involved the molecular functions such as the combination of

关 键 词：青光眼 视网膜 肝郁证 微小核糖核酸 差异表达 生物信息学 机制 miRNA-m RNA网络 

分 类 号：R276.7[医药卫生—中医五官科学]