CVB3诱导的急性心肌炎C57BL/6小鼠模型转录组分析

Transcriptomic Analysis of CVB3 Induced Acute Viral Myocarditis in C57BL/6 Mouse Model

研究柯萨奇病毒B3(CVB3)感染小鼠所引起的心肌组织转录组变化规律。C57BL/6小鼠腹腔接种浓度为104TCID50的CVB3,建立C57BL/6急性病毒性心肌炎小鼠模型,逐日测量体重。接种第3、6、9、11和14d分别取心脏,计算心脏指数,并取部分心肌组织进行HE染色分析病理学改变;病毒接种后的第3、第6和第9d,取部分组织匀浆进行转录组测序,分析三个时间点的共同差异表达基因,对其进行GO和KEGG信号通路的富集,并对其中12个基因进行了qRT-PCR的验证。在CVB3感染后,小鼠体重下降至对照组的80%,心脏指数在第3d明显升高,随后逐渐下降。通过转录组分析找到100个共同差异基因,从中选出的12个基因,经qRT-PCR验证与转录组表达趋势一致。GO和KEGG信号通路富集发现,CVB3感染后,小鼠心肌组织出现病毒性心肌炎、NK细胞通路,T、B细胞激活及先天性免疫反应通路的改变。差异基因的蛋白与蛋白互作网络分析显示,先天性免疫中MHC-Ⅰ型分子蛋白基因H2-Q7、H2-K1、H2-D1等,NK细胞毒作用通路中的Gzmb、Gzma,以及蛋白酶体信号通路基因Psmb8、Psmb9、Psmb10和lfit3位于相互作用中心。CVB3感染C57BL/6小鼠心肌组织的转录组变化涉及病毒性心肌炎、NK细胞和T、B细胞激活及先天性免疫反应等通路。CVB3感染引起的急性病毒性心肌炎是多通路和多基因综合作用的结果。

To study transcriptomic changes in myocardial tissue induced by Coxsackievirus B3(CVB3)infection.C57 BL/6 mouse model of acute viral myocarditis was established by intraperitoneal injection of CVB3(viral titer:104 TCID50).The body weight and heart mass of mice were measured at different time points.One part of the myocardial tissue was taken for HE staining3,6,9,11 and 14 days after infection to analyze pathological changes;the other was homogenized for RNA sequencing(3,6 and 9 days after infection).The genes with common changes and differentially expressed in the three time periods were selected through Venn diagram for analysis,and the GO and KEGG signaling pathways were enriched.Expression of 12 of these genes were verified by qRT-PCR.After CVB3 infection,body weight of the mice decreased significantly to 80%of that of the control group,and the heart index increased significantly on the third day and then decreased gradually.A total of 100 common differential genes were found through transcriptome analysis,from which,12 genes were verified by qRT-PCR.The results were consistent with the transcriptome analysis.Results of GO and KEGG enrichment showed that viral myocarditis,NK cell pathways,T and B cell activation and innate immune response pathways were key pathways in myocardium tissue infected with CVB3.Protein-protein interaction networks showed that MHC-Ⅰtype molecular protein genes in innate immunity,such as H2-Q7,H2-K1 and H2-D1,Gzmb and Gzma in NK cytotoxic pathway,and proteasome signaling pathway genes Psmb8,Psmb9,Psmb10 and lfit3 were the key genes involved in protein interaction.In CVB3-induced C57 BL/6 mice model,the most relevant signaling pathways involved in the transcriptome changes of myocardium tissue are viral myocarditis pathway,NK cell,T cell and B cell activation,and innate immune response pathway.Viral myocarditis is the comprehensive result of the interaction of multiple pathways and genes.