摘要
目的基于生物信息学分析与生物学实验验证的鸟嘌呤核苷酸交换因子1(VAV1)在动脉粥样硬化(AS)发生与发展过程中的表达变化,初步验证其在血管平滑肌细胞(VSMCs)中的调控作用。方法从基因表达汇编数据库(GEO)获得3组AS测序数据集,使用GEO2R、Jvenn、STRING及Cytoscape等工具鉴定差异表达基因(DEGs),并筛选中枢(Hub)基因;免疫荧光染色(IFC)检测VAV1和SMMHC的表达及定位;免疫印迹分析及实时荧光定量PCR检测VAV1、SMMHC和SM22α表达的影响;胶原凝胶收缩实验及5-乙炔基-2-脱氧尿苷(EdU)染色实验检测VSMCs收缩及增殖能力。结果66个上调基因和88个下调基因在3组数据集中均有表达变化。包括VAV1在内的10个Hub基因可能与AS进展相关。IFC实验表明VAV1在AS斑块中表达显著高于正常血管组织(P<0.05),主要定位在VSMCs中;敲低VAV1的表达显著提升VSMCs的收缩能力,抑制其增殖(P<0.05)。结论VAV1在AS斑块中表达升高,这与VSMCs收缩能力降低相关。VAV1可能是AS的潜在预后标志物及治疗靶点。
Objective To verify the expression of vav guanine nucleotide exchange factor 1(VAV1)in the progression of atherosclerosis(AS)based on bioinformatics analysis and biological experiments and,to verify its potential regulatory role in vascular smooth muscle cells(VSMCs).Methods Three datasets of AS were obtained from Gene Expression Omnibus(GEO)and differentially expressed genes(DEGs)were identified.Hub genes were screened by GEO2R,Jvenn,STRING and Cytoscape.The expression and localization of VAV1 and SMMHC in vessels were detected by immuno-fluorescence staining(IFC).Western blot and real-time PCR were used to detect the expression of VAV1,SMMHC and SM22α.The contraction and proliferation of VSMCs were detected by collagen gel contraction assay and 5-ethynyl-2 deoxyuridine(EdU)staining.Results Totally 66 up-regulated genes and 88 down-regulated genes were identified in three datasets.Screening 10 Hub genes including VAV1 may be related to the progression of AS.IFC showed that the expression of VAV1 in AS plaque was significantly higher than that in normal vascular tissue(P<0.05)and mainly located in VSMCs;Knocking down VAV1 promoted the contraction of VSMCs and inhibited its proliferation(P<0.05).Conclusions The expression of VAV1 in AS plaques is increased,which is associated with the decreased contractility of VSMCs.VAV1 is a potential prognostic biomarker and therapeutic target of AS.
作者
谢思安
徐俊玄
宁婷婷
张楠
朱圣韬
刘思
XIE Si-an;XU Jun-xuan;NING Ting-ting;ZHANG Nan;ZHU Sheng-tao;LIU Si(Department of Gastroenterology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China)
出处
《基础医学与临床》
2022年第7期1092-1098,共7页
Basic and Clinical Medicine
基金
首都医科大学附属北京友谊医院“友谊种子计划”人才项目(YYZZ202028)。
