细粒棘球蚴重组蛋白P29(rEg.P29)诱导下差异表达microRNA的筛选及分析  

Analysis of mRNA differentially expressed by a host infected with Echinococcus granulosus after immunization with the recombinant antigen P29

作　　者：朱明星[1,2,3] 张婷婷 杜先才 赵殷奇 徐士梅 杨松昊 赵巍 ZHU Ming-xing;ZHANG Ting-ting;DU Xian-cai;ZHAO Yin-qi;XU Shi-mei;YANG Song-hao;ZHAO Wei(Science and Technology Center,Ningxia Medical University,Yinchuan,Ningxia 750004,China;School of Basic Medical Science,Ningxia Medical University;Ningxia Key Laboratory of Prevention and Control of Common Infectious Diseases;School of Clinical Medicine,Ningxia Medical University)

机构地区：[1]宁夏医科大学科技中心,宁夏银川750004 [2]宁夏常见传染病防治重点实验室 [3]宁夏医科大学基础医学院 [4]宁夏医科大学临床医学院

出　　处：《中国病原生物学杂志》2021年第9期1035-1040,1046,共7页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81860366,32060805);宁夏重点研发计划项目(No.2018BEG02003,2021BEG03088);宁夏自然科学基金项目(No.2020AAC02039)。

摘　　要：目的对细粒棘球蚴重组蛋白P29(rEg.P29)诱导小鼠差异表达的microRNA进行筛选和分析,以期为rEg.P29免疫前后宿主免疫细胞的分化研究提供线索。方法将6-8周龄SPF级BALB/c小鼠随机分为2组,rEg.P29免疫组和对照组,每组12只。rEg.P29免疫组于小鼠腹部多点免疫重组蛋白100μl(含rEg.P2910μg),共免疫2次,第1次免疫2周后进行第2次免疫。提取第2周(第1次免疫后2周)、第6周(第1次免疫后4周)、第8周(第1次免疫后6周)各组小鼠外周血淋巴细胞,分离microRNA用于建库测序,利用生物信息学方法筛选rEg.P29诱导下差异表达的microRNA分子,并进行注释分析。结果测序所得数据经拼接组装比对,共获得成熟的microRNA序列342条,其中已知的microRNA序列206条,未知信息序列36条。在已知的microRNA分子中,差异表达共55个,其中上调31个,下调24个;在新发现的microRNA分子中,差异表达11个,其中上调5个,下调6个。靶基因预测结果显示,表达上调microRNA调控的靶基因有14279个,表达下调microRNA调控的靶基因13911个。GO富集注释结果显示,在上调分子中,生物学进程(BP)获得18条注释信息,其中16个microRNA调控的826个靶基因被注释到细胞分化(cell differentiation)条目下;在下调分子中,生物学进程(BP)获得21条注释信息,其中25个microRNA调控的712个靶基因被注释到细胞分化(cell differentiation)条目下。KEGG通路分析结果显示,无论是表达上调还是下调的分子,其靶基因的代谢通路均显著富集到Th1 and Th2 cell differentiation、Th17 cell differentiation、B cell receptor signaling pathway等与细胞分化相关的信号通路以及与肿瘤相关的信号通路上。结论在rEg.P29诱导下小鼠免疫细胞存在差异表达的microRNA分子,这些分子可能与免疫细胞的分化相关。Objective To screen and analyze microRNAs differentially expressed in mice in response to the recombinant antigen P29 in order to provide clues to the differentiation of host immune cells before and after immunization with rEg.P29. Methods Six-8-week-old female BALB/c mice were randomly divided into 2 groups, an rEg.P29 immunization group and a control group, with 12 mice in each group. The rEg.P29 immunization group was intraabdominally immunized twice with 100 μl of the recombinant protein(including 10 μg of rEg.P29), and the second round of immunization was performed 2 weeks after the first round. Peripheral blood lymphocytes were collected from mice in each group in week 2(2 weeks after the first round of immunization), week 6(4 weeks after the first round of immunization), and week 8(6 weeks after the first round of immunization), and microRNA was isolated and sequenced to construct a library. microRNA molecules differentially expressed in response to rEg.P29 were screened for bioinformatically, and annotation analysis was performed. Results Sequencing data were compiled and aligned. A total of 342 mature microRNA sequences were obtained, 206 of which were known microRNA sequences and 36 of which were unknown sequences. Among the known microRNA molecules, a total of 55 were differentially expressed, 31 of which were up-regulated and 24 of which were down-regulated. A total of 11 newly discovered microRNA molecules were differentially expressed, 5 of which were up-regulated molecules and 6 of which were down-regulated. Prediction of target genes indicated that 14 279 target genes were regulated by up-regulated expression of microRNAs and that 13 911 target genes were regulated by down-regulated expression of microRNAs. GO enrichment analysis of the up-regulated molecules yielded 18 annotations for biological processes(BPs). Eight hundred and twenty-six target genes that were regulated by 16 microRNAs were involved in cell differentiation. GO enrichment analysis of the up-regulated molecules yielded 21 annot

关 键 词：细粒棘球蚴 rEg.P29 MICRORNA 免疫细胞分化 

分 类 号：R383.33[医药卫生—医学寄生虫学]