Liu Bochao, Ma Xiaorui, Wang Qi, Luo Shengxue, Zhang Ling, Wang Wenjing, Fu Yongshui, Allain Jean-Pierre, Li Chengyao, Li Tingting
Department of Transfusion Medicine, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China.
Guangzhou Blood Center, Guangzhou, China.
Front Cell Infect Microbiol. 2020 Apr 21;10:135. doi: 10.3389/fcimb.2020.00135. eCollection 2020.
Common marmosets infected with GB virus-B (GBV-B) chimeras containing hepatitis C virus (HCV) core and envelope proteins (CE1E2p7) developed more severe hepatitis than those infected with HCV envelope proteins (E1E2p7), suggesting that HCV core protein might be involved in the pathogenesis of viral hepatitis. The potential role of HCV core in hepatic inflammation was investigated. Six individual cDNA libraries of liver tissues from HCV CE1E2p7 or E1E2p7 chimera-infected marmosets (three animals per group) were constructed and sequenced. By differential expression gene analysis, 30 of 632 mRNA transcripts were correlated with the immune system process, which might be associated with hepatitis. A protein-protein interaction network was constituted by STRING database based on these 30 differentially expressed genes (DEGs), showing that IL-32 might play a central regulatory role in HCV core-related hepatitis. To investigate the effect of HCV core protein on IL-32 production, HCV core expressing and mock constructs were transfected into Huh7 cells. IL-32 mRNA and secretion protein were detected at significantly higher levels in cells expressing HCV core protein than in those without HCV core expression ( < 0.01 and < 0.001, respectively). By KEGG enrichment analysis and using the specific signaling pathway inhibitor LY294002 for inhibition of PI3K, IL-32 expression was significantly reduced ( < 0.001). In conclusion, HCV core protein induces an increase of IL-32 expression via the PI3K pathway in hepatic cells, which played a major role in development of HCV-related severe hepatitis.
感染了含有丙型肝炎病毒（HCV）核心蛋白和包膜蛋白（CE1E2p7）的GB病毒B型（GBV - B）嵌合体的普通狨猴，比感染HCV包膜蛋白（E1E2p7）的狨猴患更严重的肝炎，这表明HCV核心蛋白可能参与病毒性肝炎的发病机制。研究了HCV核心蛋白在肝脏炎症中的潜在作用。构建并测序了来自感染HCV CE1E2p7或E1E2p7嵌合体的狨猴肝脏组织的六个个体cDNA文库（每组三只动物）。通过差异表达基因分析，632个mRNA转录本中的30个与免疫系统过程相关，这可能与肝炎有关。基于这30个差异表达基因（DEG），利用STRING数据库构建了蛋白质 - 蛋白质相互作用网络，表明IL - 32可能在HCV核心相关肝炎中发挥核心调节作用。为了研究HCV核心蛋白对IL - 32产生的影响，将表达HCV核心蛋白的构建体和空载体构建体转染到Huh7细胞中。在表达HCV核心蛋白细胞中检测到的IL - 32 mRNA和分泌蛋白水平显著高于未表达HCV核心蛋白的细胞（分别为<0.01和<0.001）。通过KEGG富集分析并使用特异性信号通路抑制剂LY294002抑制PI3K，IL - 32表达显著降低（<0.001）。总之，HCV核心蛋白通过PI3K途径诱导肝细胞中IL - 32表达增加，这在HCV相关严重肝炎的发展中起主要作用。
