中文摘要：

目的确定巨细胞病毒（CMV）体外能否直接攻击肝细胞,并建立肝细胞感染模型。方法①人肝细胞系感染：用人CMV（HCMV）AD169毒株感染正常人肝细胞系（L-02）和人胚肺成纤维细胞（HELF）与L-02共培养细胞,HC-MV感染的HELF细胞为阳性对照;②原代鼠肝细胞（PML）感染：用重组鼠CMV（MCMV）感染PML细胞。光镜观察细胞病变（CPE）,电镜观察病毒颗粒,间接免疫荧光（IFA）法检测HCMV抗原;原位杂交法检测MCMVIE基因。PML性质经观察其特征性超微结构和检测培养上清白蛋白水平予以鉴定。结果在PML细胞纯度〉95%（培养第8天）时感染病毒,3d后〉80%细胞出现典型CPE;电镜下胞核和胞质内见大量病毒颗粒;病毒IE基因检测呈阳性。HELF细胞可被HCMV感染;而L-02细胞无论是单独,还是与HELF共培养,其病毒标志物均呈阴性,细胞结构保持正常。结论原代鼠肝细胞体外可受到MCMV直接感染,为CMV肝炎的体外研究提供成功的肝细胞感染模型;而正常人肝细胞系不能感染HCMV,推测其表面HCMV受体发生改变或丢失。

英文摘要：

Objective To demonstrate if cytomegalovirus（CMV） could directly infect hepatocytes in vitro and develop an appropriate hepatocyte model of CMV infection.Methods Two CMV-infected liver cell models were set up.（1） Human liver cell model：Normal human liver cell line（L-02） alone and L-02 cells co-cultured with human embryo lung fibroblasts（HELF）,which were permissive for human CMV（HCMV）,were infected by AD169 strain of HCMV,respectively.HCMV infected HELF cells served as positive controls;（2） Primary murine liver cells（PMLs） model：PMLs cells were prepared from liver tissues of BALB/c mice aged 2-4 weeks and infected by murine CMV K181 strain on the 8th day after cultivation（homogeneity of hepatocytes：〉95%）,and uninfected PMLs were cultured at the same time as controls.Cytopathic effects（CPE） of all cells were observed under a light microscope（LM） every day after infection and the distribution of viral articles and the changes of cellular untrastructures under the electronic microscopy（EM）.Moreover,HCMV immediate-early antigen（IEA） and early antigen（EA） were detected by means of indirect immunofluorescent assay（IFA）.MCMV immediate early gene（IE gene） was tested by in situ hybridization（probe containing extron 2,3,4,5 of IE genes）.Results After 3 days of infection,〉80% PML cells showed classic CPE under the LM.A large number of viral particles were found in nucleoli and cytoplasm of the infected cells,and cellular ultrastructures were destroyed（as revealed by disappearance of bile canalicula and organelles） under the EM.MCMV IE genes were found in the nucleoli of the infected PMLs.In the infected HELF,the HCMV markers including classic CPE,viral articles and viral antigens were also absolutely positive.By comparison,L-02 cells,no matter cultured alone or co-cultured together with HEF,retained normal structures under the light and electron microscopes and had no any evidence of HCMV infection.Conclusion The primary murine liver cells can be di