Abstract: Objective　To investigate long non⁃coding RNA (lncRNA)⁃microRNA (miRNA)⁃messenger RNA (mRNA) interactions and identify the critical gene regulatory network during Schistosoma japonicum infections and praziquantel treatment using whole transcriptome sequencing. Methods　A total of 110 male C57BL/6 mice were randomly divided into the control group, the infection group and the treatment group. Mice in the infection treatment and the control group were infected with S. japonicum cercariae via the abdomen, and liver specimens were sampled from 10 mice 3, 6, 8 weeks post⁃infection. Praziquantel treatment was given to mice in the treatment group 8 weeks post⁃infection, and liver specimens were sampled from 10 mice 2, 4, 6, 8, 10 weeks post⁃treatment. Total RNA was isolated from mouse liver specimens, and the transcriptome library was constructed for high⁃throughput whole transcriptome sequencing. The significant differentially expressed genes were subjected to functional annotations, Gene Ontology (GO) terms enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Correlation analysis of liver specimens was performed using R Corrplot and Himsc functions, and the lncRNA⁃miRNA⁃mRNA interaction network analysis was performed using R MixOmics and Himsc functions. Results　There were 1 176 differentially expressed miRNAs, 5 270 differentially expressed mRNAs, and 2 682 differentially expressed lncRNAs between the infection group and the control group, 1 279 differentially expressed miRNAs, 7 differentially expressed mRNAs, and 69 differentially expressed lncRNAs between the treatment group and the infection group, and 1 210 differentially expressed miRNAs, 4 456 differentially expressed mRNAs, and 2 016 differentially expressed lncRNAs between the treatment group and the control group. Correlation analysis showed a higher correlation of gene expression between the treatment group and the control group. Principal component analysis showed obvious separate clustering between the infection group and the treatment group. The differentially expressed genes with significant relevance were significantly enriched in 24 GO terms, including arachidonic acid metabolic process, xenobiotic catabolic process, unsaturated fatty acid metabolic process, xenobiotic metabolic process, long⁃chain fatty acid metabolic process, and 8 KEGG metabolic pathways, including cholesterol metabolism, tyrosine metabolism, linoleic acid metabolism, retinol metabolism, and steroid hormone biometabolism. Conclusions　There were 23 mRNAs including Cyp2b9 and 14 lncRNAs including Rmrpr in the core position of the gene regulatory network, which may play a critical role in S. japonicum infections and praziquantel treatment, and 9 miRNAs including miR⁃8105 may serve as potential molecular markers for diagnosis of S. japonicum infections.

Key words: Schistosoma japonicum, Praziquantel, Whole transcriptome sequencing, Gene regulatory network, Mouse

摘要： 目的　利用全转录组测序技术研究日本血吸虫感染及吡喹酮治疗过程中长链非编码RNA（lncRNA）⁃微小RNA（miRNA）⁃信使RNA（mRNA）的交互作用，分析其中发挥调控作用的关键基因网络。方法　110只C57BL/6小鼠随机分为对照组、感染组和治疗组，感染组和治疗组以腹部贴片法感染日本血吸虫尾蚴后，第3、6、8周分别取10只小鼠肝脏组织；治疗组在第8周后开始吡喹酮治疗，治疗后第2、4、6、8、10周分别取10只小鼠肝脏组织。所有肝脏样本提取总RNA并构建转录组文库，进行全转录组高通量测序，对显著变化的差异表达基因进行功能注释、基因本体（GO）功能富集分析、京都基因与基因组百科全书（KEGG）通路富集分析。应用R语言中Corrplot包和Himsc包对肝脏标本进行相关性分析，利用R语言中的MixOmics包和Himsc包对lncRNA⁃miRNA⁃mRNA进行多组学交互网络分析。结果　感染组与对照组间共有差异表达miRNA基因 1 176个、差异表达mRNA基因5 270个、差异表达lncRNA基因 2 682个；治疗组与感染组间共有差异表达miRNA基因1 289个、差异表达mRNA基因7个、差异表达lncRNA基因69个；治疗组与对照组间共有差异表达miRNA基因1 210个、差异表达mRNA基因4 456个、差异表达lncRNA基因2 016个。相关性分析显示，治疗组与对照组基因表达相关性更高。主成分分析显示，感染组与治疗组出现了明显分别聚类。具有显著相关性的基因主要富集在花生四烯酸代谢、异生分解代谢、不饱和脂肪酸代谢、异生代谢、长链脂肪酸代谢等24个GO条目和胆固醇代谢、酪氨酸代谢、亚油酸代谢、雷丁醇代谢、类固醇激素生物代谢等8条KEGG代谢途径。结论　Cyp2b9等23个mRNA和 Rmrpr等14个lncRNA处于基因调控网络核心位置，可能在血吸虫感染及吡喹酮治疗过程中发挥关键调控作用；miR⁃8105等9个miRNA具有作为血吸虫感染诊断分子标志物的潜力。

关键词: 日本血吸虫, 吡喹酮, 全转录组测序, 基因调控网络, 小鼠