目的利用人流感病毒H1N1和H3N2感染小鼠星形胶质细胞获得条件培养上清刺激正常星形胶质细胞,研究人流感病毒感染星形胶质细胞后释放的细胞因子,是否会诱导正常胶质细胞的免疫病理学变化。方法从新生小鼠大脑皮质分离纯化星形胶质细胞,经纯度鉴定后,用人流感病毒H1N1和H3N2体外感染,分别于感染早期(6 h)和感染中晚期(24 h)收获条件培养上清,刺激正常星形胶质细胞,CCK-8法检测细胞存活率,Western blotting法检测星形胶质细胞胶质纤维酸性蛋白(GFAP)表达状况,ELISA法检测细胞不同时间分泌的炎症因子TNF-α、IL-1β、IL-6水平变化。结果流感病毒感染星形胶质细胞的条件培养上清,可显著抑制正常胶质细胞的活性,诱导GFAP蛋白表达的上调,而炎症因子TNF-α、IL-1β、IL-6在不同时间点的蛋白表达水平发生不同程度改变,总体呈下降趋势。结论体外实验中,人流感病毒感染星形胶质细胞后释放的细胞因子会诱导正常胶质细胞发生较为明显的免疫病理学变化,但未见明显的细胞因子级联现象。 OBJECTIVE: To infect astrocytes in mouse astrocytes infected with human influenza virus H1N1 and H3N2 to obtain conditional culture supernatant to stimulate normal astrocytes to study whether cytokines released from human influenza virus infected astrocytes can induce normal glue Immunopathological changes of stromal cells. Methods The astrocytes were isolated and purified from the cerebral cortex of newborn mice. After purity identification, in vitro infection with human influenza virus H1N1 and H3N2, the culture supernatants were harvested at the early stage of infection (6 h) and in the middle stage of infection (24 h) , The normal astrocytes were stimulated, the cell viability was detected by CCK-8 assay, the glial fibrillary acidic protein (GFAP) expression was detected by Western blotting, and the secretion of inflammatory cytokines TNF- α, IL-1β, IL-6 levels. Results The conditioned medium of astrocytes infected with influenza virus could significantly inhibit the activity of normal glial cells and induce the up-regulation of GFAP protein expression. The inflammatory cytokines such as TNF-α, IL-1β and IL-6 at different time points The protein expression levels changed in varying degrees, the overall downward trend. Conclusion In vitro experiments, cytokines released from human astrovirus infected human influenza virus induced more obvious immunopathological changes in normal glial cells, but no obvious cascade of cytokines was observed.