动脉型肺动脉高压分子标志基因的筛选与鉴定  

Screening and identification of molecular marker genes for pulmonary arterial hypertension

作　　者：李洁[1] 刘彩莹 严文文[1] 沈玉芹[1] 徐金媛 徐国彤[3] 吕立夏[2] 宋浩明[1] LI Jie;LIU Cai-ying;YAN Wen-wen;SHEN Yu-qin;XU Jin-yuan;XU Guo-tong;LÜLi-xia;SONG Hao-ming(Department of Cardiology,Tongji Hospital,School of Medicine,Tongji University,Shanghai 200065,China;Department of Biochemistry and Molecular Biology,School of Medicine,Tongji University,Shanghai 200092,China;Department of Pharmacology,School of Medicine,Tongji University,Shanghai 200092,China)

机构地区：[1]同济大学附属同济医院心内科,上海200065 [2]同济大学医学院生物化学与分子生物学系,上海200092 [3]同济大学医学院药学院,上海200092

出　　处：《中国病理生理杂志》2022年第6期1063-1074,共12页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金青年基金资助项目(No.81700316)。

摘　　要：目的:本研究旨在筛选和鉴定参与动脉型肺动脉高压(pulmonary arterial hypertension,PAH)发病的关键基因及相关信号通路,为进一步的转化医学研究提供新靶点。方法:从美国国立生物技术信息中心的GEO(Gene Expression Omnibus)数据库中获取人GSE113439、GSE117261、GSE48149和GSE53408基因芯片数据集,经过数据筛选后确定PAH组103例和对照组56例进行比较分析。采用NetworkAnalyst软件筛选差异基因(differentially expressed genes,DEGs),Enrichr和Metascape进行基因本体论(gene ontology,GO)和京都基因和基因组数据库(Kyo⁃to Encyclopedia of Genes and Genomes,KEGG)分析,STRING和Cytoscape建立蛋白质-蛋白质相互作用(protein-pro⁃tein interaction,PPI)网络确定潜在的枢纽基因。采用野百合碱构建PAH大鼠模型,通过测量血液动力学参数与组织形态学观察造模是否成功,RT-qPCR验证肺组织中候选基因的mRNA表达。结果:共获得2048个差异基因,其中1480个上调,568个下调,主要涉及炎症与细胞增殖等方面的信号通路,例如单纯疱疹病毒1感染通路、人乳头瘤病毒感染通路以及肿瘤通路等。其中最显著的DEGs依次为SCAMP3、MRPL11、ADRA1A、GATA2和SCAMP2,枢纽基因为MDM2、HSP90AA1、NCL、CTNNB1和PPP2CA。野百合碱诱导4周后,PAH组大鼠右心室收缩压(right ventricu⁃lar systolic pressure,RVSP)和平均肺动脉压(mean pulmonary arterial pressure,mPAP)与对照组相比显著升高(P<0.05),肺组织苏木精-伊红(hematoxylin-eosin,HE)染色显示肺小动脉管壁明显增厚(P<0.01),证明PAH模型建立成功。RT-qPCR结果表明,MDM2、HSP90AA1、NCL和CTNNB1的mRNA表达在PAH组显著上调(P<0.05),而MRPL11、ADRA1A、GATA2、BST2和PTPN11的mRNA表达在PAH组显著下调(P<0.05),SCAMP3、SCAMP2、PPP2CA、BIRC2、TAP1和GIF2的mRNA表达在两组间无显著差异(P>0.05)。结论:本研究筛选并验证了参与PAH发生的关键基因,有望为进一步的转化医学研究提供新靶点。AIM:To identify the key genes and related signaling pathways involved in the pathogenesis of pul⁃monary arterial hypertension(PAH),which can provide new targets for translational medicine research.METHODS:GSE113439,GSE117261,GSE48149 and GSE53408 gene microarray datasets were extracted from Gene Expression Om⁃nibus(GEO)database,and then 103 cases of PAH and 56 cases of healthy controls were identified after data screening for comparison analysis.NetworkAnalyst was used to screen differentially expressed genes(DEGs),and Enrichr and Metascape were used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.Moreover,protein-protein interaction(PPI)network was established using STRING and Cytoscape to identify the hub genes.Rat PAH model was constructed using monocrotaline,which was determined by measuring haemodynamic parameters and histomorphological observations.Changes in mRNA levels of candidate DEGs in lung tissues were validated by RT-qPCR.RESULTS:A total of 2048 DEGs were obtained,in which 1480 were up-regulated and 568 were down-regulated.These genes were mainly relevant to inflammation and proliferation,such as herpes simplex virus 1 infection,human papillomavi⁃rus infection and pathways in cancer.SCAMP3,MRPL11,ADRA1A,GATA2 and SCAMP2 were the most significant DEGs,and hub genes including MDM2,HSP90AA1,NCL,CTNNB1 and PPP2CA were also identified.Four weeks after the injection of monocrotaline,right ventricular systolic pressure(RVSP)and mean pulmonary arterial pressure(mPAP)of rats in PAH group were significantly higher compared to the control group(P<0.05),and hematoxylin-eosin(HE)staining of lung tissues showed that the walls of small pulmonary arteries were significantly thickened(P<0.01),which in⁃dicated PAH model was established successfully.The up-regulation of MDM2,HSP90AA1,NCL and CTNNB1,and down-regulation of MRPL11,ADRA1A,GATA2,BST2 and PTPN11 in PAH group were verified by RT-qPCR(P<0.05),while the expression of SCAMP3,SCAMP2,PPP2CA,BIRC2,TAP1 and GIF2 were not signi

关 键 词：动脉型肺动脉高压 生物信息学 炎症 细胞增殖 

分 类 号：R563[医药卫生—呼吸系统] R363.2[医药卫生—内科学]