HSV-1感染对人视网膜色素上皮细胞生长的影响

目的:建立单纯疱疹病毒1型(Herpes simplex virus,HSV-1)感染人视网膜色素上皮细胞(Retinal pigmentepith elial,RPE)的体外模型,观察HSV-1感染对体外培养RPE细胞生长的影响。方法:以感染复数(MOI)为5的HSV-1(F株)感染体外培养的RPE细胞。用相差显微镜观察RPE细胞的形态变化;用MTT法、流式细胞术观察HSV-1感染对RPE细胞增殖、凋亡和细胞周期的影响;用PCR和免疫荧光法验证HSV-1感染RPE细胞是否成功。结果:成功在体外建立了HSV-1感染RPE的细胞模型。HSV-1感染RPE细胞8h后出现病变,细胞形态开始变得细长和皱缩;感染24h后细胞病变更加明显,细胞间间隙增加,甚至可见多核巨细胞;感染48h后所有细胞均变圆,细胞开始出现脱落甚至死亡。MTT法显示HSV-1感染RPE后24h和48h能抑制细胞生长(P<0.01),细胞的抑制率分别为(16.37±3.28)%和(47.91±6.39)%;流式细胞仪检测发现HSV-1感染可影响RPE细胞的细胞周期,但对细胞凋亡无明显影响。结论:HSV-1能感染体外培养的RPE细胞,抑制RPE细胞的增殖,并影响RPE细胞的细胞周期。眼科学报2007;23:212-218.

Effects of HSV-1 on the Growth of Human Retinal Pigment Epithelial Cells

PURPOSE: The present study was aimed to investigate whether herpes simplex virus type 1 (HSV-1) could infect human retinal pigment epithelial (RPE) cells in vitro and the effects of HSV-1 on the growth of human RPE cells. METHODS: RPE cell line D407 was infected with HSV-1 F strain at a multiplicity of infection of 5. The susceptibility to HSV-1 was assessed by the detection of viral DNA using the polymerase chain reaction (PCR), and viral antigen using immunofluorescence staining. At indicated times, contrast-phase microscope was used to record the morphological changes of HSV-1 infection of RPE cells. Cell proliferation was measured by MTT assay. Cell apoptosis and cell cycle were detected by flow cytometry. RESULTS: HSV-1 infected RPE cells successfully in vitro. The contrast-phase microscope revealed that cytopathic effects (CPE) were first observed at 8 h post-infection. The infected cells became slender and shrinking. At 24 h post-infection, nearly all the cells displayed CPE changes. Occasionally multinucleated giant cells were found. At 48 h, the infected cells became round, some even detached from the culture flask. Cell proliferation were dramatically inhibited by HSV-1 at 24 h and 48 h post-infection, and the inhibition ratio were (16.37 +/- 3.28)% and (47. 91 +/- 6.39)% respectively. Furthermore, HSV-1 could perturb the cell cycle in the infected cells, and had no effect on cell apoptosis. CONCLUSIONS: Our data suggested that RPE cells are susceptible to HSV-1. HSV-1 can inhibit cell proliferation and perturb the cell cycle in the infected cells.