鸡新城疫病毒性脑炎的中枢神经系统基因表达谱的研究

发布时间：2018-10-04 21:04

【摘要】：新城疫(Newcastle disease)是由新城疫病毒(Newcastle disease virus,NDV)引起的一种急性病毒性的传染病,其特征是高热、呼吸困难、下痢和神经紊乱。NDV的宿主范围广泛,可感染250多种鸟类,在家禽中主要侵害鸡、火鸡、鹌鹑等,对养禽业造成巨大的威胁。NDV属于副黏病毒科、禽副黏病毒属成员。它是单股负链不分节段的RNA病毒,基因组全长15K核苷酸,其基因结构为3’-NP-P-M-F-HN-L-5’,分别编码6种病毒蛋白及2种非结构蛋白V和W。新城疫病毒强毒株的感染能够引起家禽非化脓性脑炎及因此导致显著的中枢神经紊乱,如扭颈、头颈震颤、转圈、腿或翅膀麻痹和瘫痪等症状,其死亡率可达90%以上。目前,对新城疫病毒神经致病机理知之甚少。本研究通过运用基因芯片技术探究新城疫病毒感染造成的家禽中枢神经系统基因差异性表达谱,力求为新城疫病毒的神经致病机理研究提供有价值的线索。本论文的主要研究结果如下:1.强弱毒力NDV感染鸡中枢神经系统的基因表达谱将4周龄的SPF鸡分别感染NDV强毒株F48E9、强毒株Yulin及弱毒疫苗株LaSota,并设立正常(未感染)对照组。感染第五天,两个强毒株组出现显著的神经症状而频临死亡,LaSota组和正常对照组没有表现任何临床症状。分别取各组鸡的脑部并进行44K鸡全基因组芯片(43,803 probes)表达谱分析。强毒株F48E9、Yulin组与对照组比较而共有的上调基因429个、下调基因267个。经过GO和KEGG pathway分析,筛选出56个重要差异表达的基因,其与神经细胞生长发育、天然免疫和细胞凋亡相关,为进一步研究提供了候选的靶基因。2.主要差异表达基因的分类筛选从差异表达显著的基因中,筛选出9个与神经细胞的生长发育有关的基因,ALDH1A2、TGIF1和YWHAG都涉及神经元的分化;ALDH1A2、MDK和CNTN4涉及神经系统发育;HRAS与NBN涉及神经元细胞的凋亡,CALB1与维持神经元细胞体的稳定有关。31个与天然免疫相关的基因,TLR4、TLR7、IRF7和IL18等与Toll样受体和干扰素调节因子有关。24个(其中8个也与天然免疫相关重叠)与细胞凋亡相关的候选基因,APAF1、BAK1和BIRC2等与诱导细胞凋亡或抑制细胞凋亡有关。这些基因可以为进一步研究NDV引起的禽病毒性脑炎提供有价值的线索。3.强弱毒力NDV的感染引起鸡脑组织的病理变化HE染色和免疫组化分析显示,强毒株F48E9和Yulin引起鸡大脑组织充血、神经元细胞软化,呈现非化脓性脑炎病理变化。此表明鸡感染NDV强毒株而导致的神经症状跟大脑组织病理变化的直接关联性。
[Abstract]:Newcastle disease (Newcastle disease) is an acute viral infectious disease caused by Newcastle disease virus (Newcastle disease virus,NDV). It is characterized by high fever, dyspnea, dysentery and neurologic disorder. Turkey, quail and so on, pose a great threat to poultry industry. NDV belongs to paramyxovirus family, avian paramyxovirus member. It is a single-strand negative strand non-segmented RNA virus with a 15K nucleotide length. Its gene structure is 3N-NP-M-F-HN-L-5, which encodes six viral proteins and two non-structural proteins V and Wrespectively. Newcastle disease virus (NDV) infection can cause non-suppurative encephalitis in poultry and thus lead to significant central nervous disorder, such as neck twisting, head and neck tremor, rotation, leg or wing paralysis and paralysis. The mortality rate is over 90%. At present, little is known about the neuropathogenetic mechanism of Newcastle disease virus (NDV). In order to provide valuable clues for the study of the neuropathogenetic mechanism of Newcastle disease virus (NDV), the differential gene expression profiles of the central nervous system caused by Newcastle disease virus (NDV) were investigated by using gene chip technique in this study. The main results of this thesis are as follows: 1. The gene expression profiles of central nervous system (CNS) infected with strong and attenuated NDV infected SPF chickens aged 4 weeks were infected with NDV virulent strain F48E9, virulent Yulin and attenuated vaccine LaSota, respectively, and were set up as normal (uninfected) control group. On the fifth day of infection, the two virulent strains showed significant neurological symptoms, while the LaSota group and the normal control group showed no clinical symptoms. The whole genome microarray (43803 probes) expression profile of 44 K chicken was analyzed. There were 429 up-regulated genes and 267 down-regulated genes in the virulent strain F48E9 Yulin group compared with the control group. By GO and KEGG pathway analysis, 56 differentially expressed genes were screened, which were related to the growth and development of nerve cells, innate immunity and apoptosis. Classification and screening of major differentially expressed genes from differentially expressed genes, Nine genes related to the growth and development of nerve cells were screened. Both ALDH1 A2TGIF1 and YWHAG were involved in the differentiation of neurons. Both ALDH1 A2MDK and CNTN4 were involved in the development of nervous system. HRAs and NBN were involved in the apoptosis of neuronal cells, CALB1 and maintaining the stability of neuronal cell bodies. 31 genes associated with innate immunity, such as TLR4, TLR7, IRF7 and IL18, are associated with Toll like receptors and interferon regulatory factors. 24 (8 of them also overlap with innate immunity) and apoptosis related candidate genes APAF1, BAK1, BIRC2, etc. Apoptosis or inhibition of apoptosis. These genes may provide valuable clues for further study of avian viral encephalitis caused by NDV. The pathological changes of chicken brain tissue caused by the infection of strong virulence NDV and immunohistochemical analysis showed that F48E9 and Yulin caused congestion and neuronal cell softening in chicken brain tissue and presented pathological changes of non-suppurative encephalitis. This suggests a direct correlation between neurologic symptoms and pathological changes in brain tissue in chickens infected with NDV virulent strains.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S855.3

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