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目的研究水痘-带状疱疹病毒(VZV)在人神经母细胞瘤细胞株(SHSY5Y)中培养及增殖的特点,观察VZV感染SHSY5Y细胞后的生物学效应。方法于普通倒置显微镜和电子显微镜下观察被感染细胞的形态变化;利用CCK-8法检测VZV感染对细胞增殖的影响;比较不同感染复数(MOI)和不同浓度胎牛血清维持液对病毒增殖的影响。结果普通倒置显微镜下观察细胞病变效应(cytopathic effect,CPE),从24h~96h,细胞病变数量增加和病变程度加重。电镜观察显示:感染48 h后核染色质固缩、浓染,线粒体肥大及增生,胞质空泡化和自噬小体形成,可见包装成熟的病毒颗粒。CCK-8法提示VZV感染24h后细胞增殖被抑制,各实验组存活率与对照组比较差异均有统计学意义(P<0.05)。在2%胎牛血清的维持液中用MOI为1的VZV感染SHSY5Y细胞,能收获较高滴度的病毒。结论初步建立了VZV感染SHSY5Y细胞的体外模型,并利用该模型观察病毒感染对细胞生物性状的影响,为探讨VZV的致病及潜伏机制奠定了基础。
Objective To investigate the characteristics of varicella - zoster virus (VZV) cultured and proliferated in human neuroblastoma cell line SHSY5Y and observe the biological effects of VZV infection on SHSY5Y cells. Methods The morphological changes of infected cells were observed under inverted microscope and electron microscope. The effects of VZV infection on cell proliferation were detected by CCK-8 assay. The effects of different concentrations of MOI and different concentrations of fetal bovine serum on virus proliferation were compared influences. Results Under normal inverted microscope, the cytopathic effect (CPE) was observed. From 24 h to 96 h, the number of cytopathic lesions and the severity of lesions were increased. Electron microscopy showed that after 48 hours of infection, nuclear chromatin condensation, staining, mitochondrial hypertrophy and hyperplasia, cytoplasmic vacuolization and autophagy formation were observed. The CCK-8 method suggested that the cell proliferation was inhibited after VZV infection for 24 h, and there was significant difference between the experimental groups and the control group (P <0.05). Infection of SHSY5Y cells with VZV at a MOI of 1 in a 2% fetal bovine serum maintenance solution yielded higher titers of virus. Conclusion The in vitro model of VZV infection in SHSY5Y cells was established and the effect of virus infection on cell biological characteristics was observed. This study laid the foundation for exploring the pathogenesis and latent mechanism of VZV infection.