摘要
目的筛选铁蓄积大鼠食管黏膜组织差异表达基因,并分析差异表达基因的生物学功能。方法12只6周龄雄性SD大鼠随机分为铁蓄积组和对照组,每组6只,铁蓄积组隔天腹腔注射蔗糖铁溶液制备铁蓄积模型,对照组注射等量生理盐水,14周后取血和食管、肝等组织,应用血清学、组织学方法鉴定铁蓄积造模是否成功;剥离两组食管黏膜组织,采用转录组测序技术筛选差异表达基因,并对差异表达基因进行基因本体(GO)功能富集分析及真核生物蛋白相邻类的聚簇(KOG)分类富集分析。结果筛选出9个差异表达基因,5个上调基因包括昼夜相关转录抑制因子(Ciart)、液泡蛋白质分选因子25(Vps25)、甲状腺激素应答蛋白(Thrsp)、UDP-N-乙酰氨基葡萄糖焦磷酸化酶1(Uap1)、血清解整合素—金属蛋白酶33(Adam33),4个下调基因包括过氧化物酶基因(Pxdn)、硫酸乙酰肝素蛋白多糖基因2(Hspg2)、中心体相关蛋白2(Cep2)、G蛋白信号转导调节因子4(Rgs4)。GO功能富集分析显示,上调基因的生物过程(BP)集中在节律过程、代谢过程、行为、生物过程调节、特定位置运动、生物过程的负调控、生物调节、定位、细胞过程、多细胞生物过程;细胞组成(CC)主要集中在膜封闭腔、细胞器部分、细胞器、膜、细胞外区域部分、膜部分、细胞部分、细胞;分子功能(MF)主要集中在结合、结构分子活性、催化活性。下调基因的BP主要集中在解毒、刺激反应、细胞组成或生物形成、信号、生物过程的负调控、生物过程的正调节、发育过程、细胞过程、生物过程调节、生物调节、代谢过程;CC主要集中在细胞外基质、细胞外基质组成、细胞外区域部分、细胞外区域、细胞器、膜、细胞部分、细胞;MF主要集中在酶调节活性、抗氧化活性、分子功能调节剂、受体调节活性、结构分子活性、结合、催化活性。KOG分类富集分析显示,表达上调的基因中有3个获得功能注释,Uap1被注释到细胞壁/膜/包膜生物形成,Adam33被注释到翻译后修饰,蛋白质折叠和伴侣蛋白,Vps25被注释到功能预测。表达下调的基因中有2个获得功能注释,Rgs4被注释到信号转导机制,Hspg2被注释到翻译后修饰,蛋白质折叠和伴侣蛋白。结论铁蓄积大鼠食管黏膜组织中共筛选出9个差异表达基因,包括5个表达上调基因和4个表达下调基因。GO功能富集分析显示差异表达基因主要参与的BP包括节律过程、代谢过程、细胞过程、生物过程调节等,主要参与的CC包括细胞器、膜、细胞外区域部分等,主要参与的MF包括结合、催化活性、结构分子活性。有5个差异基因获得KOG功能注释,包括翻译后修饰、蛋白质折叠和伴侣蛋白、信号转导等,主要参与细胞周期、细胞代谢、细胞增殖及氧化应激等通路。
Objective To screen the differentially expressed genes(DEGs)in the esophageal mucosa of rats with iron accumulation and to analyze the biological function of DEGs.Methods Twelve 6-week-old male Sprague-Dawley rats were divided into the iron accumulation group(n=6)and control group(n=6).Rats in the iron accumulation group received bi-daily injections of sucrose iron solution,while rats in the control group received the equal volume of normal saline.After 14 weeks,the blood,esophagus and other organs were collected,and histological and serological techniques were employed to confirm weather modeling was successful or not.Esophageal mucosa was dissected for transcriptome sequencing,and DEGs were screened and annotated.The functional annotations of DEGs were conducted by Gene Ontology(GO)and the classification annotations of DEGs were conducted by clusters of orthologous groups for eukaryotic complete genomes(KOG).Results A total of 9 DEGs were screened;5 significantly up-regulated genes included circadian-asso-ciated repressor of transcription(Ciart),vacuolar protein sorting 25 homolog(Vps25),thyroid hormone-responsive pro-tein(Thrsp),UDP-N-acetylglucosamine pyrophosphorylase 1(Uap1),and ADAM metallopeptidase Domain 33(Ad-am33);4 significantly down-regulated genes included peroxidasin(Pxdn),heparan sulfate proteoglycan 2(Hspg2),cen-trosomal protein 2(Cep2)and regulator of G-protein signaling 4(Rgs4).GO enrichment analysis showed that the biologi-cal process(BP)of up-regulated genes was mainly concentrated in rhythmic process,metabolic process,behavior,regula-tion of biological process,establishment of localization,negative regulation of biological process,biological regulation,lo-calization,cellular process,and multicellular biological process;the cellular component(CC)was mainly concentrated in membrane-enclosed lumen,organelle part,organelle,membrane,extracellular region part,membrane part,cell part,cell;the molecular function(MF)was mainly concentrated in binding,structural molecular activity,and catalytic activi-ty;the BP of down-regulated genes was mainly concentrated in detoxification,response to stimulus,cellular component or-ganization or biogenesis,signaling,negative regulation of biological process,positive regulation of biological process,de-velopmental process,cellular process,regulation of biological process,biological regulation,and metabolic process;the CC was mainly concentrated in extracellular matrix,extracellular matrix component,extracellular region part,extracellu-lar region,organelle,membrane,cell part,and cell;the MF was mainly concentrated in enzyme regulation activity,anti-oxidant activity,molecular function regulator,receptor regulator activity,structural molecular activity,binding,and cata-lytic activity.KOG enrichment analysis showed that 3 of the up-regulated genes were annotated,Uap1 was annotated to cell wall/membrane/envelope biogenesis,Adam33 was annotated to posttranslational modification,protein turnover,chap-erones,and Vps25 was annotated to general function prediction only,while 2 of the down-regulated genes were annotated,Rgs4 was annotated to signal transduction mechanisms,and Hspg2 was annotated to posttranslational modification,protein turnover,and chaperone.Conclusions A total of 9 DEGs were screened out,including five significantly up-regulated genes and four significantly down-regulated genes.GO enrichment showed that the BP of DEGs included rhythmic pro-cess,metabolic processes,cellular process,and regulation of biological processes,etc.,CC of DEGs included organelle,membrane,extracellular region,and cell,etc.,MF of DEGS included binding,catalytic activity,and structural molecu-lar activity,etc.KOG enrichment showed that five DEGs were annotated with KOG functions,including posttranslational modification,protein turnover,chaperones,signal transduction and others.These DEGs were mainly involved in the path-ways including cell cycle,cell metabolism,cell proliferation,and oxidative stress,etc.
作者
刘国红
任雨轩
邵谦毅
梁硕
王丽萍
LIU Guohong;REN Yuxuan;SHAO Qianyi;LIANG Shuo;WANG Liping(School of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450001,China;不详)
出处
《山东医药》
CAS
2024年第12期37-41,共5页
Shandong Medical Journal
关键词
铁蓄积
差异表达基因
基因本体功能富集
真核生物蛋白相邻类的聚簇分类富集
iron accumulation
differentially expressed genes
gene ontology
clusters of orthologous groups for eukaryotic complete genomes
