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目的定量研究汉滩病毒(HTNV)体外诱导细胞热休克蛋白70(HSP70)及HSP70 mRNA的表达规律。方法以HTNV敏感细胞株Vero-E6为对象,体外实验性感染Hantaan 76-118。采用原位杂交、免疫细胞化学染色法及逆转录聚合酶链反应(RT-PCR)观察感染后72 h内HSP70的表达并加以定量分析。结果与模拟感染组相比,Hantaan 76-118感染Vero-E6细胞后0．5 h．HSP70 mRNA开始升高,于感染后12 h达高峰,并持续高表达至72 h(P＜0．05)。Hantaan 76-118感染后2 h可检测到HSP70蛋白表达增加,于感染后12 h达高峰,并持续高表达至72 h(P＜0．05)。结论体外感染HTNV可诱导Vero-E6细胞产生热休克反应并表达HSP70,这种快速、持久的应激反应,可能对HSP70持续发挥其细胞保护作用具有重要意义。
Objective To quantitatively study the expression of heat shock protein 70 (HSP70) and HSP70 mRNA induced by Hantaan virus (HTNV) in vitro. Methods HTNV sensitive cell line Vero-E6 was used to infect Hantaan 76-118 in vitro. In situ hybridization, immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR) were used to observe the expression of HSP70 within 72 hours after infection and quantitative analysis. Results Compared with the mock-infected group, Hantaan 76-118 infected with Vero-E6 cells at 0.5 h. HSP70 mRNA began to rise and peaked at 12 h after infection, and persisted to high level for 72 h (P <0.05). Hantaan 76-118 2 h after infection can be detected HSP70 protein expression increased at 12 h after infection peaked and continued to high expression to 72 h (P <0.05). Conclusion In vitro infection of HTNV induced heat shock response and expression of HSP70 in Vero-E6 cells. This rapid and long-lasting stress response may play an important role in protecting HSP70 from cytoprotection.