感染肺炎支原体姜曲海猪肺组织差异表达circRNAs的筛选与分析  

Screening and Analysis of Differentially Expressed circRNAs in the Lung Tissue of Jiangquhai pigs Infected with Mycoplasma hyopneumoniae

作　　者：倪黎纲[1,2] 赵旭庭[1] 王宵燕[2] 徐盼[1] 甘源[3] 金文[1] NI Ligang;ZHAO Xuting;WANG Xiaoyan;XU Pan;GAN Yuan;JIN Wen(Jiangsu Argi-Animal Husbandry Vocational College,Taizhou 225300, China;College of Animal Science and Technology, Yangzhou University,Yangzhou225009,China;Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210004, China)

机构地区：[1]江苏农牧科技职业学院,泰州225300 [2]扬州大学动物科学与技术学院,扬州225009 [3]江苏省农业科学院兽医研究所,南京210004

出　　处：《中国畜牧兽医》2021年第6期2212-2220,共9页China Animal Husbandry & Veterinary Medicine

基　　金：江苏现代农业产业技术体系建设项目(JATS[2020]346);江苏农牧科技职业学院院级课题(NSF201802)。

摘　　要：为探明姜曲海猪感染猪肺炎支原体(Mycoplasma hyopneumoniae,Mhp)后肺组织环状RNA(circular RNA,circRNA)差异性表达谱及其在抗Mhp感染中的作用,试验以姜曲海猪为研究对象,分为感染组和对照组,人工感染Mhp 28 d后,解剖采集肺组织,采用高通量测序和生物信息学软件分析circRNA的表达情况。测序数据比对参考猪基因组序列,共鉴定到23632个circRNAs,差异表达circRNAs为213个,其中97个上调,116个下调。随机选择4个差异表达circRNAs进行实时荧光定量PCR验证,检测结果与测序结果基本一致。差异表达circRNA来源基因可注释到包括抗原加工递呈、溶酶体、白细胞跨内皮迁移等免疫应答信号通路。circRNA-miRNA-mRNA靶标关系分析显示,筛选到海绵结合miRNA数量最多的3个差异表达circRNAs,分别为:circRNA-17284(21个)、circRNA-04848(19个)和circRNA-17270(19个);预测到6个与免疫调控相关的靶向miRNAs,分别为:ssc-miR-4331、ssc-miR-370、ssc-miR-328、ssc-miR-30c-3p、ssc-miR-122和ssc-miR-125b。本研究测定了感染Mhp的猪肺组织circRNA表达谱,筛选到与免疫调控相关的差异表达circRNA,这有助于阐明姜曲海猪对Mhp的易感机制,为抗病育种研究提供了参考依据。In order to investigate the differential expression profile of circular RNA(circRNA)in the lung tissue of pigs infected with Mycoplasma hyopneumoniae(Mhp)as well as its role in Mhp infection.Jiangquhai pigs were used as subjects in this experiment,which were divided into Mhp-infected and control groups.On the 28th day post inoculation,the lung tissues were collected from all of the experimental pigs for analysis of circRNA expression by high-throughput sequencing and bioinformatics software.Data comparison showed that a total of 23632 circRNAs were identified,with 213 differentially expressed circRNAs(DEcircRNAs).Among them,97 up-regulated and 116 down-regulated.Four from 213 DEcircRNAs were randomly selected and verified by Real-time quantitative PCR,and the results were consistent with those obtained by sequencing.Host genes of DEcircRNAs were annotated to immune-related signaling pathways,including antigen processing and presentation,lysosome and leukocyte transendothelial migration.The target relationship analysis of circRNA-miRNA-mRNA showed that three DEcircRNAs with the highest number of sponge miRNAs were circRNA-17284(21),circRNA-04848(19)and circRNA-17270(19),of which 6 targeted miRNAs related to immune regulation were predicted,including ssc-miR-4331,ssc-miR-370,ssc-miR-328,ssc-miR-30c-3p,ssc-miR-122 and ssc-miR-125b.In this study,the expression profile of circRNA in the lung tissue of pigs infected with Mhp was determined,and the DEcircRNAs related to immune regulation was screened,which helped to clarify the mechanism of susceptibility of Jiangquhai pigs to Mhp infection,and provide scientific basis for disease-resistant breeding research.

关 键 词：姜曲海猪 猪肺炎支原体 环状RNA 高通量测序 

分 类 号：Q786[生物学—分子生物学]