缺血性脑卒中两种血瘀证的生物信息学分析及实验验证  

Bioinformatics Analysis and Experimental Validation of Two Blood Stasis Patterns in Ischemic Stroke

作　　者：梁晨曦 李筱璐 陈建敏 王柯文 桂裕昌 张钰琴[2] 许建文[2] LIANG Chenxi;LI Xiaolu;CHEN Jianmin;WANG Kewen;GUI Yuchang;ZHANG Yuqin;XU Jianwen(The First College of Clinical Medicine,Guangxi Medical University,Nanning 530021,China;The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一临床医学院,南宁530021 [2]广西医科大学第一附属医院,南宁530021

出　　处：《世界科学技术-中医药现代化》2024年第10期2695-2702,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：广西壮族自治区中医药管理局广西中医药重点学科建设项目(GZXK-Z-20-52):中西医结合临床学科,负责人:吕军影。

摘　　要：目的探索缺血性脑卒中(IS)气虚血瘀(QDBS)和阴虚血瘀(YDBS)证型的差异表达基因(DEGs)及相关信号通路,从分子水平阐明IS血瘀证可能的发生机制及生物学基础。方法从GEO数据库下载大鼠IS两个证型的mRNA表达谱数据集GSE100235,利用R语言的limma包筛选DEGs。然后使用DAVID数据库进行GO和KEGG分析。联合使用miRDB和miRWalk数据库预测DEGs的miRNA,构建miRNA-mRNA调控网络。随后使用STRING数据库构建蛋白-蛋白互作网络,采用cytoHubba插件识别核心靶基因。分别采用力竭游泳训练结合改良Longa线栓法构建QDBS大鼠模型,氢化可的松注射及线栓法构建YDBS大鼠脑缺血模型。RT-qPCR法验证两组大鼠脑组织中核心靶点的表达水平。结果共筛选出47个DEGs,主要涉及神经元凋亡、炎症反应、适应性免疫反应等生物过程,参与脂质和动脉粥样硬化和C型凝集素受体信号通路。miRNA-mRNA互作网络包括64个节点和55条边。共获得5个核心靶基因,分别为Ccl2、Selp、Timp1、Ccr1l1和Fpr1。结论IS大鼠QDBS和YDBS证型中存在显著差异表达的基因,涉及多种生物过程和作用途径,其中与脂质和动脉粥样硬化信号通路联系最为密切。Objective To explore the differential expression genes(DEGs)and related signaling pathways of the Qi deficiency and blood stasis(QDBS)and Yin deficiency and blood stasis(YDBS)syndromes in ischemic stroke(IS)at the molecular level,elucidating the potential mechanisms and biological basis of blood stasis syndrome in IS.Methods The mRNA expression profile dataset GSE100235 of the two syndromes of IS rats was downloaded from the GEO database,and the limma package of R language was used to screen DEGs.Subsequently,GO and KECG analyses were performed using the DAVID database.The miRDB and miRWalk databases were used to predict the miRNAs of DEGs,and aa miRNA-mRNA regulatory network was constructed.A protein-protein interaction network was built using the STRING database,and the cytoHubba plug-in was utilized to identify the core target genes.The rat model of QDBS in IS was established by exhaustive swimming training combined with modified Longa's thread embolism,and the rat model of YDBS in IS was established by hydrocortisone injection and thread embolism.RT-qPCR was employed to validate the expression levels of core target genes in rat brain tissues of the two groups.ResultsA total of 47 DEGs were identified.These genes were mainly involved in biological processes such as neuronal apoptosis,inflammatory response,and adaptive immune response,as well as pathways related to lipid metabolism,atherosclerosis,and C-type lectin receptor signaling.The miRNA-mRNA interaction network consisted of 64 nodes and 55 edges.Five core target genes were obtained,including Ccl2,Selp,Timpl,Ccr1l1,and Fprl.Conclusion There are significantly differentially expressed genes in QDBS and YDBS syndrome of IS rats.These genes are involved in a variety of biological processes and pathways,and are most closely related to lipid metabolism and atherosclerosis signaling pathways.

关 键 词：缺血性脑卒中 GEO数据库 生物信息学 气虚血瘀证 阴虚血瘀证 

分 类 号：R277.7[医药卫生—中医学]