流感病毒对MDCK细胞核转录因子表达的影响

Effect of influenza virus on the expression of MDCK cellular nucler transcription factors in vitro

目的研究流感病毒刺激对体外培养的MDCK细胞核转录因子表达的影响,探讨流感病毒损伤细胞的分子学机制。方法体外培养获得MDCK细胞,分别用NF-κB免疫组化方法和TransAM P65试剂盒测定血凝滴度1:64的A型流感病毒H1N1和H3N2两种亚型、B型流感病毒、紫外线照射灭活的B型流感病毒NF-κB蛋白表达和活性的动态变化。以细胞激活剂佛波酯刺激后的MDCK细胞为阳性对照,以等量生理盐水为阴性对照。结果各组细胞加入不同的流感病毒后,与对照组相比,细胞免疫组化显示NF-κB随流感病毒作用相同时间后结果出现明显差异;各组相比,B型流感病毒作用72h对NF-κB的激活作用最明显,而A型两种亚型之间无显著差异。TransAM P65试剂盒测定也得出相同的结果。结论流感病毒可激活体外培养MDCK细胞的核转录因子,且这种作用随时间而增强。

Objective To investigate the impact of influenza virus A subtypes H1N1/H3N2 and influenza virus B as well as inactivated influenza virus NF-κB uhravilet-radiation on the expression of NF-κB in MDCK cells in vitro and prelimina-rily explore the molecular mechanism of cells breakdown by influenza virus. Methods The MDCK ceils were harvested by in vitro culture. They were grouped by exposed samples. The activity and the expression of proteins of influenza virus A sub-type H1N1/H3N2 and influenza virus B as well as influenza virus NF-κB B p65 protein inactivated by ultravilet-radiation in MDCK cells in vitro were determined with immunohistochemistry and TransAM P65 kit. Results In cells exposed to influenza virus, P65 transferred from cytoplasma to cell nucleus. The magnitude was different to kinds of samples. Type B influenza virus was capable of activating NF-κB most obviously. Conclusion Influenza can activate NF-κB in MDCK ceils cultured in vitro, and this kind of activating action is correlated with the exposure time.