塔里木马鹿损伤与正常茸组织的转录组特征差异分析  

Analysis of the transcriptome characteristics of damaged and normal antler tissues of the Cervus elaphus yarkandensis

作　　者：刘嘉伟 陈红 王淼 贾小东 方翟[3] 韩春梅[1,2] LIU Jiawei;CHEN Hong;WANG Miao;JIA Xiaodong;FANG Di;HAN Chunmei(College of Animal Science and Technology,Tarim University,Aral 843300,China;Key Laboratory of Tarim Animal Husbandry Science and Technology of Xinjiang Production Construction Corps,Aral 843300;College of Life Sciences and Technology,Tarim University,Aral 843300,China)

机构地区：[1]塔里木大学动物科学与技术学院,新疆阿拉尔843300 [2]新疆生产建设兵团塔里木畜牧科技重点实验室,新疆阿拉尔843300 [3]塔里木大学生命科学与技术学院,新疆阿拉尔843300

出　　处：《黑龙江畜牧兽医》2023年第12期130-136,148,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目“创伤微环境HGF/c-Met信号对鹿茸间充质干细胞增殖、迁移与分化的调控机制”(31860629);塔里木大学科研创新项目“基于转录组测序技术筛选马鹿鹿茸再生的关键基因”(TDGRI202134)。

摘　　要：为了探究损伤鹿茸的快速修复生长机制,挖掘与修复生长相关的候选基因,试验首先采集塔里木马鹿损伤后修复53 d(损伤组)与正常生长53 d(正常组)的茸组织,提取总RNA后,基于Illumi-na Nova Seq 6000高通量测序平台进行转录组测序,将过滤后序列与参考基因组的序列比对后筛选差异表达基因(differentially expressed genes,DEGs);然后对DEGs进行GO功能富集分析与KEGG信号通路富集分析;最后随机选取4个DEGs(IHH、IL2RA、BVES和LOC122686105基因)进行实时荧光定量PCR分析,验证转录组测序结果的可靠性。结果表明:损伤组与正常组之间存在56个DEGs;与正常组比较,损伤组有37个DEGs显著上调(|lb Fold Change|≥1,P<0.05),包括IHH、IL2RA、BVES、Wnt5b、COL9A2、CRMP5等,有19个DEGs显著下调(|lb Fold Change|≥1,P<0.05),包括LOC122686105、ALX3、VWF等;损伤组有23个显著上调(P<0.05)的GO二级条目,包括生物过程(biological process,BP)中的19个GO二级条目[骨骼系统发育(GO:0001501)、软骨发育(GO:0051216)等]、细胞成分(cellular component,CC)中的2个GO二级条目[肌纤维膜(GO:00042383)、闰盘(GO:0014704)]和分子功能(molecular function,MF)中的2个GO二级条目[蛋白结合(GO:0005515)、相同蛋白质结合(GO:0042802)],以及8个显著下调(P<0.05)的GO二级条目,包括BP中的7个二级条目[细胞外区域(GO:0005576)、化学稳态(GO:0048878)、骨骼生长(GO:0098868)等]和CC中的1个二级条目[蛋白的N端结合(GO:0047485)];损伤组有13条显著上调(P<0.05)的KEGG信号通路,包括Hedgehog信号通路、Wnt信号通路、类固醇的生物合成等,有9条显著下调(P<0.05)的KEGG信号通路,包括维生素的消化吸收、胆固醇代谢和PPAR信号通路等。在56个DEGs中,与鹿茸修复生长相关的候选基因有IHH、IL2RA、Wnt5b、COL9A2、CRMP5。经实时荧光定量PCR分析发现,与正常组比较,损伤组中IHH、IL2RA、BVES基因显著上调表达(P<0.05),LOC122686105基因显著下调表�In order to explore the rapid repair and growth mechanism of damaged deer antler and explore candidate genes related to repair growth,the antler tissues of the Cervus elaphus yarkandensis were collected at 53 days after injury repair(injured group)and 53 days after normal growth(normal group).After total RNA was extracted,transcriptome sequencing was performed based on Illumina Nova Seq 6000 high-throughput sequencing platform.The filtered sequences were compared with the reference genome sequences and differentiated expression genes(differentially expressed genes,DEGs)were screened.GO functional enrichment analysis and KEGG signaling pathway enrichment analysis were performed on DEGs.Finally,four DEGs were randomly selected for real-time fluorescence quantitative PCR analysis to verify the reliability of transcriptome sequencing results.The results showed that there were 56 DEGs between injured group and normal group.Compared with the normal group,37 DEGs in the injured group were significantly up-regulated(|lb Fold Change|≥1,P<0.05),including IHH,IL2RA,BVES,Wnt5B,COL9A2,CRMP5,etc.,and 19 DEGs were significantly down-regulated(|lb Fold Change|≥1,P<0.05),including LOC122686105,ALX3,VWF,etc.In the injured group,there were 23 GO level 2 items that were significantly up-regulated(P<0.05),including 19 GO secondary entries(skeletal phylogeny[GO:0001501],chondrogenesis[GO:0051216],etc.)of biological process(BP),2 GO secondary entries(fibromyofilm[GO:00042383],leap disk[GO:0014704])of cellular component(CC)and 2 GO secondary entries(protein binding[GO:0005515],same protein binding[GO:0042802])of molecular function(MF),and 8 GO secondary entries that were significantly down-regulated(P<0.05),including 7 GO secondary entries(including extracellular region[GO:0005576],chemical homeostasis[GO:0048878]of BP and bone growth[GO:0098868])and 1 secondary entry(N-terminal binding of protein[GO:0047485])of CC.In the injured group,13 KEGG signaling pathways were significantly up-regulated(P<0.05),including Hedgehog signaling pa

关 键 词：塔里木马鹿 鹿茸组织 损伤修复 转录组测序 差异表达基因(DEGs) 

分 类 号：S874[农业科学—畜牧兽医]