单纯疱疹病毒Ⅰ型感染胎鼠原代神经元细胞模型的建立

Establishment of primary cultured neurons model of mouse embryo with HSV-Ⅰ infection

目的 建立单纯疱疹病毒Ⅰ型 (HSV Ⅰ )感染胎鼠原代皮质神经元细胞模型。方法 取孕 14～ 16d昆明小鼠皮质神经元进行细胞培养并感染HSV Ⅰ。观察倒置光学显微镜下神经元形态变化 ,并应用流式细胞仪检测二者的百分比。应用间接免疫荧光方法观察正常及病毒感染组神经元和星形胶质细胞的变化。用特异性HSV Ⅰ荧光抗体检测神经元受染情况 ,并利用MTT比色实验检测细胞活性。结果 原代培养神经元数量、形态良好。用阿糖胞苷抑制星形胶质细胞生长后 ,神经元纯度较高。培养 3d时加入HSV Ⅰ感染神经元 ,HSV Ⅰ特异性免疫荧光反应阳性 ,受染神经元形态变化 ,活性降低。结论 HSV Ⅰ可直接感染原代培养神经元细胞 。

Objective To establish a neurons model for herpes simplex virus type 1 (HSV Ⅰ) infection using cultures of mouse embryo Methods The cortical neuron of 14 16 day old mouse embryo were cultured and infected with HSV Ⅰ Microscopic examination, immunofluorescent test, MTT colorimetric analysis and flowcytometric assay were used to detect the immunofluorescent reaction of normal and infected neurons and astrocytes, respectively The percentage of the two kinds of cells were measured Result The mouse neurons could proliferate well in vitro The neurons amount met the requirement of cytological experiment after treatment with Ara C in the tissue culture HSV Ⅰ could infect the neurons directly, the activity of the infected neurons was obviously reduced Conclusion HSV Ⅰ could successfully infect the primary cultured neurons The model may provide a useful approach for studying the pathogenesis of herpes simplex virus encephalitis