黄芩苷体外干扰IBV在CEK细胞内复制的转录组分析
Transcriptome analysis of baicalin interfering with IBV replication in CEK cells in vitro
郭霄慧 1李卫晴 1王转转 1刘一宁 1李伟欣 1陈光明 1冯婉莹 1贾青辉 1张召兴2
作者信息
- 1. 河北科技师范学院河北省预防兽医学重点实验室,河北秦皇岛 066604
- 2. 河北科技师范学院河北省预防兽医学重点实验室,河北秦皇岛 066604;河北旅游职业学院,河北承德 067000
- 折叠
摘要
为了解黄芩苷干扰禽传染性支气管炎病毒(IBV)感染鸡胚肾(CEK)细胞内复制前后细胞基因表达的差异性及表达情况,进一步揭示和分析黄芩苷体外干扰IBV在CEK细胞内复制的作用机制,IBV感染CEK细胞2 h后,加入9.75 mg/L的黄芩苷药液进行干扰,为黄芩苷(H-IBV)组,同时设立细胞对照(Cell)组和IBV感染(IBV)组各3个复孔,培养36 h后收集细胞样品进行有参转录组测序.结果H-IBV组与IBV组相比共有102个显著差异表达基因,其中显著上调基因48个,显著下调基因54个;通过基因本体论(GO)和京都基因与基因组百科全书(KEGG)数据库中的功能注释和通路富集发现,差异表达基因主要注释在细胞过程、生物调节、代谢等生物过程和病毒性传染病、信号转导与相互作用等二级通路中,其中CEK细胞内视黄醇代谢通路、磷脂转移到膜、IL-27介导的信号通路、MDA5/RIG-I和Toll样受体信号通路均显著富集,并且Ⅰ型干扰素及干扰素α的产生、抗病毒感染过程也均得到了正向调节,富集到核酸催化、免疫系统与反应和生物间相互反应的差异基因数较多;通过STRING网络互作图发现大部分免疫相关基因能够组成一个有36个节点且以IRF7、TLR3、STAT1为中心的相互作用网络.故黄芩苷作用后与IBV组对比的差异表达基因主要注释和富集在生物过程中,免疫系统与反应也被加强,通过正向调节MDA5/RIG-I受体信号通路中的IRF7和抑制Toll样受体信号通路中的TLR3信号转导为主,正调IL-27介导的通路和调控JAK-STAT等信号通路为辅,激活IRF7、TLR3、STAT等相关基因并且与对应的下游蛋白相互作用,促进IFN-α和调控细胞因子的表达,再加上对病毒的(防御)反应和病毒过程的负调控,从而起到干扰IBV在CEK细胞内复制的作用.
Abstract
In order to understand the difference and expression of genes in CEK cells before and af-ter baicalin interferes with IBV,further reveal and analyze the mechanism of baicalin interfering IBV replication in CEK cells in vitro.After IBV infected CEK cells for 2 h,9.75 mg/L baicalin liq-uid was added to interfere with CEK cells,which was recorded as the baicalin(H-IBV)group,and three replicate wells were set in the control group and the IBV(IBV)infection group.After 36 h culture,cell samples were collected and subject to transcriptome for sequencing.The results showed that there were 102 differentially expressed genes in H-IBV group compared with IBV in-fection group,among which 48 genes weresignificantly up-regulated and 54 genes were significantly down-regulated.Through functional annotation in GO and KEGG databases,it was found that dif-ferentially expressed genes were mainly annotated in biological processes such as cellular proces-ses,biological regulation,metabolism,and secondary pathways such as viral infectious diseases,signal transduction and interaction.Retinol metabolism pathway,phospholipid transfer to mem-brane,IL-27 mediated signaling pathway,MDA5/RIG-I and Toll-like receptor signaling pathway were significantly enriched in CEK cells,and the production of type Ⅰ interferon and interferon al-pha and the process of antiviral infection were also positively regulated.There were more differen-tial genes enriched in nucleic acid catalysis,immune system,and reaction,and interbiological reac-tion.Through the STRING network interaction map,it was found that most immune-related genes could form a 36-node interaction network centered on IRF7,TLR3 and STAT1.Therefore,com-pared with IBV group,the differentially expressed genes after baicalin treatment were mainly an-notated and enriched in the biological process,and the immune system and response were en-hanced,mainly through the positive regulation of IRF7 in the MDA5/RIG-I receptor signaling pathway and the inhibition of TLR3 signal transduction in the Toll-like receptor signaling path-way.Positive regulation of IL-27 mediated pathway and regulation of JAK-STAT signaling path-way were supplemented by activation of the expression of IRF7,TLR3,STAT1 and other related genes,and interaction with corresponding downstream proteins to promote the expression of IFN-α and regulatory cytokines,coupled with negative regulation of viral(defense)response and viral processes.Thus,baicalin interferes with IBV replication in CEK cells.
关键词
黄芩苷/鸡传染性支气管炎病毒/鸡胚肾(CEK)细胞/转录组Key words
baicalin/avian infectious bronchitis virus/CEK cells/transcriptome引用本文复制引用
基金项目
河北省重点研发计划(21326614D)
河北省现代农业产业技术体系建设项目蛋禽创新团队项目(HBCT2024260209)
河北省现代农业产业技术体系建设项目肉禽创新团队项目(HBCT2024270207)
河北省预防兽医学重点实验室绩效后补助资助项目(20567621H)
出版年
2024
NETL
NSTL
万方数据