miR-1249在BVDV诱导MDBK细胞凋亡与持续性感染发生过程中的机制研究

牛病毒性腹泻病毒（BVDV）的持续性感染对于该病的防治至关重要，但致病机制目前尚不清楚。miRNA已被证实在病毒感染过程中起到重要调控作用。BVDV致病机制是否与miRNA有关？我们前期研究表明BVDV感染细胞后miR-1249表达显著上调，并可促进细胞凋亡。提示：miR-1249可能与BVDV持续性感染有关。为进一步阐明miR-1249调控病毒感染的分子机制，明确其在病毒感染中的作用，本项目拟首先利用高通量测序技术建立BVDV感染MDBK细胞的转录本文库，利用该文库信息和生物信息学技术预测miR-1249的调亡相关靶基因，并用荧光素酶报告系统等技术进行验证。通过qPCR、流式细胞技术等方法研究miR-1249在细胞凋亡和BVDV感染中的调节作用。本研究将从miRNA的角度揭示BVDV引起持续性感染的机制，同时为寻找新的抗病毒治疗方法和新的抗病毒治疗靶点提供理论依据。

Mechanisms of persistent infection induced by bovine viral diarrhea virus (BVDV) is essential for the prevention and treatment of Mucosal disease. The pathogenic mechanisms are concerned, but it is not yet clear. Indeed, some research have reported that host-cellular miRNAs, modulating the expression of various viral genes, play an important role in the host-pathogen interaction network. miRNA become possibly a breakthrough for elucidating mechanisms of BVDV infection. Our previous studies showed that miR-1249 was up-regulated significantly. miR-1249, as class of modulators, may play an important role in the regulation of cell apoptosis pathway which is involved in BVDV persistent infection. To investigate the regulatory role of miR-1249 in BVDV infection and the underlying mechanisms, total RNAs from cpBVDV-infected MDBK cells and ncpBVDV-infected MDBK cells at different time points were extracted and then subjected to Transcriptome Resequencing. Target genes of miR-1249 will be predicted by using gene array technique and target-predicting software such as TargetScan. Target genes of miR-1249 will be validated by using luciferase reporter system. Regulatory role of miRNAs will be investigated in MDBK apoptosis and persistent infection induced by BVDV using RT-PCR and Flow cytometry technology. The study will determined the role of miRNA in MDBK apoptosis and persistent infection by BVDV, which lay a foundation for developing novel anti-BVDV therapeutic strategies.