摘要
细胞内源性microRNA(miRNA)在宿主与病原相互作用过程中发挥着重要的调节功能。为了分析细胞内源性miRNA与传染性法氏囊病病毒(IBDV)的相互作用,用IBDV弱毒和强毒分别感染鸡胚成纤维细胞(CEF)和SPF鸡,24h后,取感染CEF和法氏囊组织,提取细胞总RNA,用Hy3/Hy5双色荧光标记,与miRNA芯片杂交,进行芯片内标准化、芯片间标准化、表达差异比较以及聚类分析。结果显示:在IBDV弱毒感染的CEF中,有17个细胞内源性miRNA表达上调,17个miRNA表达下调;在IBDV强毒感染的鸡法氏囊组织细胞中,有30个细胞内源性miRNA表达上调,18个miRNA表达下调。根据表达差异显著的miRNA序列设计引物,用荧光定量RT-PCR方法验证芯片检测结果,2种方法的检测结果一致。结果表明,IBDV感染可诱导细胞内源性miRNA表达变化,这些上调或下调的miRNA能调控细胞内多种代谢和信号传导途径发生异常,引起细胞及组织器官发生病理学变化。
Abstract： MicroRNAs （miRNAs） mark a new paradigm of RNA-directed gene expression regulation in a wide spectrum of biological systems. These small non-coding RNAs can contribute to the repertoire of host-pathogen interactions during viral infection. This interplay has important consequences, both for the virus and the host. There have been reported evidences of host-cellular miRNAs modulating the expression of various viral genes,thereby playing a pivotal role in the host-pathogen interaction network. To explore the function of the host-cellular miRNA in the chick embryo fibroblast （CEF） and specific pathogen free（SPF）chicken infected with IBDV respectively,we compared the miRNA expression profiles in normal CEF and CEF infected with low virulent IBDV after 24 hours, and normal SPF chicken and SPF chicken infected with highly virulent IBDV after 24 hours respectively, using a microrray containing more than 1 700 capture probes,covering all microRNAs annotated in miRBase 11.0,as well as all viral microRNAs, related to these species. The results indicated that 17 miRNAs were downregulated in CEF infected with IBDV after 24 hours compared to normal CEF, 17 miRNAs upregulated. In SPF chicken groups, 30 miRNAs were detected downregulated,18 miRNAs upregulated. The microarray results were verified by quantitative real-time PCR（qRT-PCR）. Our experiment provided the data to further expound the molecular mechanism of differential expression of host-cellular miRNA involved in the pathogenesis of CEF and SPF chicken infected with IBDV.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2012年第3期329-336,共8页
Chinese Journal of Veterinary Science
基金
江苏省自然科学基金资助项目(BK2009041
BK2010471)
中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室开放基金资助项目(SKLVBF201105)
