【摘要】 目的：研究确定人巨细胞病毒（ Ｈ Ｃ Ｍ Ｖ） 体内外先天性感染胎鼠大脑皮质神经元及其感染特征。方法：分别建立体内外模拟 Ｈ Ｃ Ｍ Ｖ 先天性感染小鼠脑神经元模型。采用病毒分离、病理学检测及核酸原位杂交技术等观察研究病毒的增殖，受染细胞的形态特征以及病毒核酸在细胞内的定位和分布。结果：体内模型显示： 脑组织上清液中分离出 Ｈ Ｃ Ｍ Ｖ； 大脑皮质发生侵袭性脑膜脑炎性病理改变； 神经元核内出现病毒特征性大的嗜碱性包涵体；病毒特异性核酸存在于受染神经元核内及胞浆内。体外模型亦证实：受染神经元胞体明显肿胀并可见核内嗜酸性包涵体； 原位杂交法检出 Ｈ Ｃ Ｍ Ｖ特异性核酸。结论： Ｈ Ｃ Ｍ Ｖ 可在体内外致胎鼠大脑皮质神经元发生类似于人类先天性中枢神经系统感染的病理改变； 建立的体内外感染模型可用于 Ｈ Ｃ Ｍ Ｖ 中枢神经系统先天性感染的发生机制研究。更多还原

【Abstract】 Objective : To investigate is to determine the infection of neurons that human cytomegalovirus A D169strain( H C M V A D169)could cross the placenta ofthe Balb/cmice and initiate congenitalinfection of the developing fetuscerebral cortexinvivo and thatthe neurons of the primary cultures from the cerebral cortex of Balb/c fetal mice could beinfected by H C M V A D169 in vitro . Methods : H C M V(5.0 log T C I D50 ml/ mouse)was injected into the intraperitoneum ofmice (half of mice are female) when they were about 8 ～12 weeks old . Then , these of mice were used for mat ing . Pregnancies were dated as day 0 when a copulation plug was found . Fetuses on day about to give birth of gestationwere removed from the uteri and its cerebral cortex was removed from the skull and fixed with 4% paraformaldehyde,thencutinto serial coronal sections 5 μm thick,applied to slides coated with Histostik ,and then stained with hematoxylin andeosin ( H E) . The supernatant of minced a fetal mouse cerebral cortex was used for virusisolates by H Fcells . And viral D N Apositive cells were examined byin situ hybridization using digoxigenin labelled H C M V D N Aoligonucleotide probe inthe acetone fixed imprint ofthe cerebral cortex. Meanwhile , The cerebral cortex neurons were prepared and cultured and H C M Vinfection model was made . 1 ～4 weeks afterinfection , the cultures was recorded and observed by photographyunder microscope , and the samples were taken for H Estainning. Atthe same time ,the virus D N Aand its intracellularlocation were analyzed in situhybridization ～using digoxingenin labelled H C M V D N A oligonucleotide probes. Results : H C M Vhad been isolafed from the supernatant of fetal mice cerebral cortex. The pathological changes consisting of de structive meningoencephalitis andlarge 、basophilic 、intranuclear neuronalinclusions of viraltype occurred in the fetal mousecerebral cortex . The presence of virus sequences was confirmed byin situ hybridization ,however,nothing was found in thenormal controls . The neurons exhibited cytopathic effects at 2 weeks of postinfection , and the viral sequences was foundwhen they were infected by H C M V A D169 in vitro . Conclusion : Developmentofthe fetal mouse model of H C M Vinfectionthat mimics human congenital central nervous system( C N S) infection in vivo and in vitro will allow the study of thepathogenesis of H C M Vcongenital C N Sinfection and the development oftherapeutic agents and vaccines.[更多还原