人巨细胞病毒感染人胚肺成纤维细胞对其细胞周期的影响

目的研究人巨细胞病毒（human cytomegalovirus,HCMV）感染人胚肺成纤维细胞（human embryoniclung fibroblast,HELF）后对其细胞周期的影响,探讨HCMV感染的发病机制。方法用HCMV感染同步化的HELF作为感染组,同时设模拟感染组为对照组。用倒置相差显微镜观察两组细胞形态学变化至感染后7d。于感染后12h、24h、48h、72h和96h收获细胞,用免疫细胞化学法检测两组细胞核内HCMVIE172蛋白,流式细胞术检测细胞周期。结果模拟感染组未见细胞形态学改变,亦未出现HCMVIE172蛋白阳性产物。感染组感染后72h时即可见局部细胞变圆,膨胀,胞体及胞核巨大化,7d时可见所有细胞均发生病变;免疫细胞化学检测显示,其各时间点的细胞均在核内出现呈棕黄色颗粒的HCMVIE172蛋白。模拟感染组在感染后12h时有65.7%的细胞处于G1期,24h时有43.8%的细胞处于G1期;感染组在感染后12h时有70.4%的细胞处于G1期,24h时有69.9%的细胞处于G1期;两两比较,差异均有显著性（均P〈0.01）。结论HCMV感染HELF后在细胞核内进行复制、增殖,其影响了HELF周期,使大部分细胞停滞在G期。

Effect of human cytomegalovirus infection on cell cycle of human embryonic lung fibroblast

Objective To study the effects of human cytomegalovirus(HCMV) infection on cell cycle of human embryonic lung fibroblast(HELF),and explore the pathogenesis of HCMV infection.Methods Synchronized HELF were infected with HCMV,the simulant-infected group was set up as control group simultaneously.Observation of the pathomorphological change of infected HELF was performed until 7 days of postinfection by inverted phase contrast microscope.Infected HELF cells were harvested at 12,24,48,72 and 96 hours of post-infection.HCMV IE172 protein in nucleus was detected by immunocytochemistry method,cell cycle of HELF was detected by flow cytometry analysis.Results The change in morphology was not observed and positive products of HCMV IE172 protein were not detected in the simulant-infected cells.In HCMV-infected group,the partial cells became round,bulgy,and their cytoplasm and nucleus became large at 72 hours of post-infection,cytopathic effect was observed in all infected cells at 7 days of post-infection.Immunocytochemistry test showed the brown and yellow granule appeared at all time points in HCMV-infected cells.At 12 hours of post-infection,65.7% of the cells were in G1 phase in the simulant-infected group,but 70.4% of the cells were in G1 phase in the HCMV-infected group;at 24 hours of post-infection,43.8% of the simulant-infected cells were in G1 phase,but 69.9% of the HCMV-infected cells were in G1 phase.The statistical analysis suggested that there were significant difference(P<0.01) in the number of cells in G1 phase between the simulant-infected group and HCMV-infected group both at 12 and 24 hours of post-infection.Conclusion HCMV can replicate and propagate in HCMV-infected HELF,HCMV infection influences the progression of cell cycle of HELF and arrests cell cycle at G1 phase.