热休克蛋白70在寨卡病毒感染中的作用  

The role of heat shock protein 70 in Zika virus infection

作　　者：刘巧巧 赖宏智 刘婷 蒲洁莹[1,2,3,4] 黄曦 LIU Qiao-qiao;LAI Hong-zhi;LIU Ting;PU Jie-ying;HUANG Xi(Institute of Immunology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080;Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai , Guangdong 519000;Institure of Human Virology, Sun Yatsen University, Guangzhou, Guangdong 510080;Key Laboratory of Tropical Disease Control Research of Ministry of Education, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080;Third People Hospital, Shantou, Guangdong 515073, China)

机构地区：[1]中山大学中山医学院免疫学教研室,广东广州510080 [2]中山大学第五附属医院,广东珠海519000 [3]中山大学人类病毒学研究所,广东广州510080 [4]中山大学热带病防治研究教育部重点实验室,广东广州510080 [5]汕头市第三人民医院,广东汕头515073

出　　处：《热带医学杂志》2018年第6期703-707,共5页Journal of Tropical Medicine

基　　金：国家自然科学基金(31470877;31370868;81261160323)

摘　　要：目的探讨热休克蛋白70(Hsp70)在寨卡病毒(ZIKV)感染中的作用。方法建立ZIKV感染的A549细胞模型,Realtime-PCR方法检测感染后不同时间点Hsp70表达水平;在A549细胞中过表达Hsp70-flag或沉默Hsp70,感染ZIKV后,Realtime-PCR方法检测ZIKV RNA表达水平和肿瘤坏死因子(TNF-α)、干扰素-β(IFN-β)表达水平,Western blot方法检测细胞中病毒蛋白的表达水平;用Hsp70的抑制剂VER155008处理A549细胞,Realtime-PCR方法检测ZIKV RNA表达水平和TNF-α、IFN-β表达水平,Western blot方法检测病毒蛋白表达水平,并用FocusForming Assay检测病毒粒子释放水平。结果 A549细胞感染ZIKV后,Hsp70表达上调,感染组是未感染组的2倍(P<0.05);在A549细胞中过表达Hsp70-flag后,ZIKV复制水平和TNF-α、IFN-β水平均增高(P<0.05);在A549细胞中沉默Hsp70后感染ZIKV,病毒胞内复制减弱,TNF-α、IFN-β的表达水平降低(P<0.05);抑制Hsp70的功能后,ZIKV胞内复制水平降低,TNF-α、IFN-β表达水平降低(P<0.05)。结论 Hsp70可促进ZIKV在A549细胞中的增殖及促炎性因子TNF-α、IFN-β的释放,并且这种作用依赖于Hsp70与ATP结合的功能域。Objective To investigate whether heat shock protein 70（Hsp70）plays a role in Zika virus（ZIKV）infection.Methods A549 cells were infected with Zika virus（SZ01）;the expression levels of Hsp70 at different time points weredetected by Realtime-PCR.A549 cells were transfected with Hsp70-flag plasmid or si RNA,followed by Zika virus infection;the expression levels of Zika virus RNA and proteins were evaluated by Realtime-PCR and Western blot,respectively.Theexpression levels of tumor necrosis factor-α（TNF-α）,interferon-β（IFN-β）were also detected.Hsp70 in A549 cells wereinhibited followed by/after Zika infection;the expression levels of Zika virus RNA and proteins were detected by Realtime-PCRand Western blot,respectively;TNF-α and IFN-β were also detected.The releasing of virus particles was tested by FocusForming Assay.Results The expression level of Hsp70 in A549 cells was upregulated by 200% after Zika infection（P〈0.05）.After transfection with Hsp70-flag in A549 cells,the expression levels of Zika virus was upregulated,as well as TNF-α andIFN-β（P〈0.05）.After transfection with si RNA in A549 cells,the expression level of Zika virus was downregulated,as well asTNF-α and IFN-β（P〈0.05）.Inhibition of Hsp70 in A549 leads to reduction of Zika virus paticles and TNF-α and IFN-β（P0.05）.Conclusion Hsp70 could promote the infection of Zika virus in A549 cells,which was dependent on its ATP bindingregion.

关 键 词：HSP70 寨卡病毒 肿瘤坏死因子 干扰素-Β 

分 类 号：R373[医药卫生—病原生物学]