中国实验动物学报2023，Vol.31Issue(12)：1519-1529,11.DOI:10.3969/j.issn.1005-4847.2023.12.001
C57BL/6N-Tg(1.28HBV)/Vst乙肝病毒转基因小鼠的转录组学分析
Transcriptomic analysis of C57BL/6N-Tg(1.28HBV)/Vst hepatitis B virus transgenic mice
摘要
Abstract
Objective To observe the characteristics of C57BL/6N-Tg(1.28HBV)/Vst transgenic hepatitis B virus(HBV-Tg)model mice and analyze their transcriptomic characteristics.Methods Twenty male HBV-Tg mice were divided into an experimental group and a wild-type(control)group(n = 10 mice per group).The virological characteristics of the model mice were evaluated according to serum levels of HBV DNA,HBsAg,and HBeAg,and expression levels of HBsAg and HBcAg in liver tissue.Serum levels of alanine transaminase(ALT)and aspartate transaminase(AST),hematoxylin and eosin(HE)and Sirius red staining,and hydroxyproline(Hyp)in liver tissue were detected to evaluate the degree of liver inflammation and fibrosis.Liver tissue samples were randomly selected from three mice in each group for RNA extraction for high-throughput transcriptome sequencing.Significantly differentially expressed genes were identified using R software.Functional enrichment of differential genes was determined by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses,and genes with significant differences were verified by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results ALT and AST levels were increased in the model group compared with the normal group,with the result for ALT being more significant(P<0.05).HE staining of liver tissue showed enlargement of the liver nucleus and swelling of some hepatocytes in the model group,while Sirius red staining showed a small amount of collagen deposition in the sink area and interlobule in the HBV transgenic group,in the shape of thin lines.A total of 1352 differential genes were obtained by screening(|logFC|>2 and P.adj<0.05),including 703 up-regulated and 649 down-regulated genes.KEGG analysis suggested that differential genes were mainly enriched in the peroxisome proliferator-activated receptor(PPAR)signaling pathway,retinol metabolism,fatty acid degradation,and other pathways(P.adj<0.05).The main significantly up-regulated genes included Cyp4a10,Cyp4a14,Acot1,Acot3,and Ehhadh,and the significantly down-regulated genes included Scn5a、Apol10b、Igddc4、Cxcl1、9530077C05Rik.The trend was consistent after RT-qPCR detection(P<0.05).Conclusions HBV-Tg mice have a tendency to develop spontaneous fibrosis.Transcriptomic analysis showed that chronic hepatitis B mainly involves PPAR signaling,retinol metabolism,fatty acid degradation,drug metabolism,and other pathways.关键词
乙型病毒性肝炎/转基因小鼠/转录组学/基因表达Key words
chronic hepatitis B/transgenic mice/transcriptomics/gene expression分类
生物科学引用本文复制引用
李容容,孙鑫,黄恺,赵志敏,彭渊,刘成海..C57BL/6N-Tg(1.28HBV)/Vst乙肝病毒转基因小鼠的转录组学分析[J].中国实验动物学报,2023,31(12):1519-1529,11.基金项目
国家自然科学基金(82205052,82174057,82274305),上海市科委启明星计划扬帆专项(22YF1449300),上海中医药大学预算内项目(2021LK081),上海市临床重点专科建设项目(shslczdzk01201),山西中医药大学科技创新团队项目(2022TD2003). Funded by National Natural Science Foundation of China(82205052,82174057,82274305),Shanghai Science and Technology Commission Sailing Project(22YF1449300),Shanghai University of Traditional Chinese Medicine Budget Project(2021LK081),Shanghai Key Clinical Specialty Construction Project(shslczdzk01201),Science and Technology Innovation Team Project of Shanxi University of Traditional Chinese Medicine(2022TD2003). （82205052,82174057,82274305）
