PCV2感染猪肺泡巨噬细胞自噬过程中miRNA差异表达谱及靶基因功能调控网络研究

项目名称： PCV2感染猪肺泡巨噬细胞自噬过程中miRNA差异表达谱及靶基因功能调控网络研究

项目编号： No.31302071

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 王小敏

作者单位： 江苏省农业科学院

项目金额： 22万元

中文摘要： 猪肺泡巨噬细胞（PAM）不仅是PCV2的靶细胞,而且是重要的免疫细胞。PCV2可以感染PAM、不引起凋亡，逃避细胞免疫识别引起免疫抑制，而自噬可抑制凋亡产生。病毒可通过miRNA干扰宿主细胞的正常免疫过程，实现免疫逃逸。因而，我们设想PCV2感染可能会引起miRNA调控的改变和巨噬细胞发生自噬，从而抑制凋亡产生，逃避免疫，引起免疫抑制。本课题前期研究表明，PCV2感染PAM可引起自噬现象。因此，拟以猪肺泡巨噬细胞系为模型，探讨PCV2-自噬-PAM之间的关系；对自噬过程中的PAM进行miRNA差异表达分析；预测差异表达miRNA的靶基因并进行验证，绘制靶基因调控网络，确立调控病毒在肺泡巨噬细胞中复制和自噬的miRNA；通过miRNA与靶基因mRNA相互作用的功能学验证，确立miRNA对靶基因生物学功能的影响；验证miRNA是否抑制PAM凋亡，探索PCV2免疫逃避机制，具有一定的理论意义和应用价值。

中文关键词： 猪圆环病毒2型；自噬；microRNA；靶基因；基因功能

英文摘要： Porcine alveolar macrophages （PAM） is not only the target cells of PCV2 but also an important immune cells. PCV2 could infect PAM, not result in apoptosis, escape the cellular immune identify and cause immunosuppression. Autophagy can inhibit apoptosis and virus can escape cell-mediated immunity by miRNA interfere with the normal immune process of host cells. We assumed that PCV2 infection may lead to changes of miRNA regulation and autophagy of alveolar macrophages, thereby inhibiting apoptosis, evading the immune system and causing immunosuppression. The results of our preliminary study indicated that PCV2 infection could cause autophagy on the alveolar macrophages. Therefore, we use the porcine alveolar macrophage cell lines as a model to explore the relationship between PCV2, autophagy and PAM. miRNA microarray is used to detect differential expression of miRNA in the autophage process of PCV2-infected PAM. The way of biological information is used to predict the target gene. And the target gene is tested and verified, then construct the regulatory networks between the specific expression of microRNA and the target gene to get the critical miRNA which regulated replication of PCV2 on PAM. The interaction between the miRNA and target gene is validated to analyze the impact of miRNA on the biological function

英文关键词： porcine circovirus type 2；autophagy；microRNA；target gene；gene