CAJ | 학술논문

目的建立小鼠巨细胞病毒(MCMV)先天感染模型,并观察其脑组织的病理变化及感染状况.方法取孕11～13.5d的BALB/c鼠,模型组羊膜腔微量接种K181毒株(病毒量1×103 PFU),对照组注射DMEM培养液.单独饲养5d后处死孕鼠,剖宫取出胎鼠,麻醉处死,取胎鼠脑组织制作冰冻切片.胎脑组织切片常规HE染色,光镜下观察病理学变化；采用免疫酶组化及免疫荧光法检测脑组织中的MCMV早期抗原.结果感染组胎鼠存活率为71.9％.与对照组相比,羊膜腔内接种MCMV病毒对胎鼠存活率、吸收胎率、死胎率及胎鼠头部重量无明显影响,但可致胎鼠体重降低.感染组胎鼠脑组织出现明显的病理变化.在宫内感染组,免疫酶学组化法发现脑室区、脑室管膜下区、大脑皮质和海马区可见病毒感染细胞；免疫荧光法观察到的主要感染部位与免疫酶组化法基本一致.结论通过羊膜腔内注射MCMV病毒成功建立MCMV先天感染小鼠模型,为研究MCMV先天感染致脑发育异常机制提供合适的整体研究模型.
목적 건립소서거세포병독(MCMV)선천감염모형,병관찰기뇌조직적병리변화급감염상황.방법 취잉11～13.5d적BALB/c서,모형조양막강미량접충K181독주(병독량1×103 PFU),대조조주사DMEM배양액.단독사양5d후처사잉서,부궁취출태서,마취처사,취태서뇌조직제작빙동절편.태뇌조직절편상규HE염색,광경하관찰병이학변화；채용면역매조화급면역형광법검측뇌조직중적MCMV조기항원.결과 감염조태서존활솔위71.9％.여대조조상비,양막강내접충MCMV병독대태서존활솔、흡수태솔、사태솔급태서두부중량무명현영향,단가치태서체중강저.감염조태서뇌조직출현명현적병리변화.재궁내감염조,면역매학조화법발현뇌실구、뇌실관막하구、대뇌피질화해마구가견병독감염세포；면역형광법관찰도적주요감염부위여면역매조화법기본일치.결론 통과양막강내주사MCMV병독성공건립MCMV선천감염소서모형,위연구MCMV선천감염치뇌발육이상궤제제공합괄적정체연구모형.
Objective To establish the murine congenital infection model by MCMV and observe the pathological changes and infection status of brain tissue.Methods After anesthesia,mice who were pregnant 11-13.5 days (E11-13.5 d) were intra-amniotic injected one uterus by one with virus (MCMV K181 suspension,1 μl,1×103 PFU).The control group of the same period was intra-anmiotic injected with culture medium DMEM (1 μl).Carefully reset the uteruses and close the abdomen.After 5 days of separated feeding,kill the pregnant mice,take the fetus out of the uterus,anesthetize and kill them.Make frozen sections of these fetal brains.Some sections were stained using conventional HE method,to observe the pathological changes under the light microscope.Detect MCMV early antigen in the brain tissue by immunohistochemistry staining and immunofluorescence assay.Results The survival rates of the infected group were 71.9％.Compared with the control group,intra-amniotic inoculation of MCMV does not affect the rate of fetal survival,fetal absorption,fetal death and the average weight of the heads,but decrease their average weight of the bodies.The pathological changes are found in the brain tissue of the mouse in the infection group.Through enzyme immunohistochemistry assay,there are many MCMV infected cells in brain-ventricular zone,brain subependymal zone,cerebral cortex and hippocampus area in the infection group.Similar findings were observed by immunofluorescence method.Conclusion By intra-amniotic injection of MCMV suspension,murine model of MCMV congenital infection can be successfully established.This model could be used to study the mechanisms of encephalodysplasia caused by congenital CMV infection in vivo.