林麝不同组织miRNA表达谱  

The expression profile of miRNA in the different tissues of forest musk deer(Moschus berezovskii)

作　　者：余苗杰 徐忠鲜 蒋雪梅 王春花 赵婵娟[4] 戚文华 竭航[5] YU Miaojie;XU Zhongxian;JIANG Xuemei;WANG Chunhua;ZHAO Chanjuan;QI Wenhua;JIE Hang(College of Life Science and Engineering,Chongqing Three Gorges University,Chongqing 404100,China;College of Life Sciences,China West Normal University,Nanchong 637009,China;College of Animal Medicine,Hunan Agricultural University,Changsha 410125,China;Chongqing Three Gorges Vocational College,Chongqing 404155,China;Chongqing Institute of Medicinal Plant Cultivation,Chongqing 408435,China)

机构地区：[1]重庆三峡学院生物与食品工程学院,重庆404100 [2]西华师范大学生命科学学院,南充637009 [3]湖南农业大学动物医学院,长沙410125 [4]重庆三峡职业学院,重庆404155 [5]重庆市药物种植研究所,重庆408435

出　　处：《兽类学报》2023年第6期723-733,共11页Acta Theriologica Sinica

基　　金：国家自然科学基金(82274046,32370560);重庆市自然科学基金(cstc2019jcyj⁃msxmX0410,CSTB2023NSCQ⁃MSX0123);中国博士后科学基金(2017M612959)。

摘　　要：林麝(Moschus berezovskii)是药用经济动物,其雄麝分泌的麝香是中成药和香水的重要成分,具有极高的药用价值和经济价值。miRNAs在动物细胞发育、分化、凋亡及疾病发生发展过程中起着至关重要的作用,并且在不同组织中miRNAs选择性表达。本研究利用RNA⁃seq技术在Hi⁃seq 2500测序平台对林麝的心脏、肝脏、脾脏、肺脏、肾脏和肌肉组织进行高通量测序,运用生物统计学和生物信息学方法分析这6种器官和组织中miRNAs的表达情况,采用RT⁃qPCR对测序结果进行验证。结果表明,林麝的心脏、肝脏、脾脏、肺脏、肾脏和肌肉组织中共表达的miRNAs有1650个,特异表达的miRNAs分别有86个、54个、44个、68个、83个和50个(P<0.01),共表达的miRNAs在不同器官和组织中表达水平存在显著差异(P<0.05),林麝心脏中部分特异miRNAs的表达水平远远高于其他5种器官和组织中特异miRNAs的表达水平。KEGG分析表明:林麝肝脏特异表达miR⁃NA靶基因主要涉及Wnt信号通路(ko04310)、cAMP信号通路(ko04024)、ECM受体相互作用(ko04512)、Notch信号途径(ko04330)、AMPK信号通路(ko04152)等(P<0.01)。随机选择9个miRNAs,利用RT⁃qPCR验证其在健康和化脓性林麝肝脏中的表达情况,结果表明其中9个miRNAs的表达趋势与测序结果一致。Forest musk deer(Moschus berezovskii)is a medicinal economic species and the musk secreted by the male musk deer is an important component of Chinese patent medicine and perfume,which has high medicinal and economic value.miRNAs play an important role in animal cell development,differentiation,apoptosis,and disease development,which are selectively expressed in different tissues.The heart,liver,spleen,lung,kidney,and muscle tissues of forest musk deer were high-throughput sequenced by using RNA-seq technology on the Hi-seq 2500 sequencing platform.The microRNA expression of these 6 tissues was analyzed by using biostatistics and bioinformatics methods and RT-qPCR was used to verify the sequencing results.The results showed that 1650 miRNAs co-expressed in the heart,liver,spleen,lung,kidney,and muscle tissues of forest musk deer,of which there were 86,54,44,68,83,and 50 specific miRNAs,respectively(P<0.01).The expression levels of co-expressed miRNAs were significantly different in these six types of tissues(P<0.05)and the expression levels of some specific miRNAs in the heart were much higher than that in other tissues.KEGG analysis showed that the target genes of specific miRNAs in the liver of forest musk deer were closely related to the Wnt signaling pathway(ko04310),cAMP signaling pathway(ko04024),ECM-receptor interaction(ko04512),Notch signaling pathway(ko04330),and AMPK signaling pathway(ko04152)(P<0.01).Nine miRNAs were randomly selected for verification with RT-qPCR in liver tissue of health and abscess forest musk deer and the results were consistent with the sequencing results.

关 键 词：MICRORNA 表达谱 表达分析 荧光定量PCR 林麝 

分 类 号：Q752[生物学—分子生物学]