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目的探讨单纯疱疹病毒Ⅱ型(HSV-2)诱导人宫颈上皮(HCE)细胞上调细胞的自噬活性和表达水平的作用。方法实验分为HSV-2感染组、HSV-2瞬转组及不加病毒处理的对照组,通过Western blot分析、直接免疫荧光、间接免疫荧光、质粒DNA的转染等方法研究HCE细胞在正常培养条件下的自噬活性、HSV-2感染HCE细胞的自噬活性水平及激活自噬则使HCE对HSV-2感染诱导的白细胞介素-6(IL-6)和β干扰素(IFN-β)表达的影响。结果与He La和HEK293T细胞相比,HCE细胞具有高基础水平的自噬;HSV-2感染HCE后进一步诱导上调细胞的自噬活性和表达水平(P<0.05),并且呈时间依赖性的变化。自噬特异性抑制剂显著降低HSV-2感染所诱导的IL-6和IFN-β的表达(P<0.05),自噬激活剂则促进HSV-2感染诱导的IL-6和IFN-β的表达(P<0.05)。结论 HCE细胞有着高基础水平的自噬;HSV-2感染HCE细胞早期进一步上调自噬活性水平;HSV-2感染HCE细胞诱导的自噬存在时间依赖性的变化规律;自噬参与了HSV-2感染HCE细胞的天然免疫应答。
Objective To investigate the effect of HSV-2 on the up-regulation of autophagy activity and expression in human cervical epithelial (HCE) cells. Methods The experiment was divided into HSV-2 infection group, HSV-2 transient infection group and control group without virus treatment. Western blot analysis, direct immunofluorescence, indirect immunofluorescence and plasmid DNA transfection were used to investigate the effect of HCE cells in normal Autophagy activity of HSV-2-infected HCE cells and activation of autophagy led HCE to inhibit HSV-2-induced interleukin-6 (IL-6) and interferon- β) expression. Results HCE cells had a high basal level of autophagy compared with HeLa and HEK293T cells. HSV-2 induced a further upregulation of autophagy activity and expression (P <0.05) after HCE infection, and showed a time-dependent change . The autophagy-specific inhibitor significantly decreased the expression of IL-6 and IFN-β induced by HSV-2 infection (P <0.05), while the autophagy-specific inhibitor promoted the expression of IL-6 and IFN-β induced by HSV-2 infection Expression (P <0.05). Conclusions HCE cells have high basic level of autophagy. HSV-2 infected HCE cells further upregulate the level of autophagy activity; HSV-2 infected HCE cells induced by time-dependent changes in autophagy; autophagy involved in HSV-2 The natural immune response to HCE cells.