| 魏容,肖光军,刘艳婷,等.肠道病毒 71型感染的星形胶质细胞中 EV71 3C活性与 NF-κB信号转导效率的关联[J].安徽医药,2021,25(10):2022-2027. |
| 肠道病毒 71型感染的星形胶质细胞中 EV71 3C活性与 NF-κB信号转导效率的关联 |
| Correlation between EV-type 3C protease activity and NF-κB signal transduction efficiency in astrocytes infected with enterovirus 71 |
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| DOI：10.3969/j.issn.1009-6469.2021.10.026 |
| 中文关键词: 肠道病毒感染 星形细胞 EV71 3C蛋白 转化生长因子 β活化激酶 -1 核因子 -κB信号通路 |
| 英文关键词: Enterovirus infections Astrocytes EV71 3C protein TGF -β activated kinase-1 NF-κB signal pathway |
| 基金项目:遂宁市科技计划项目( 2015s15) |
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| 中文摘要: |
| 目的探究肠道病毒 71型感染的星形胶质细胞中肠病毒 71(EV71)3C蛋白酶活性与核因子 κB(NF-κB)信号转导效率的关联。方法本研究起止时间为 2018年 3月至 2019年 3月。通过将 EV71病毒感染星形胶质细胞后，经过质粒提取、转化、定点突变、基因转变等步骤，将 TANK结合激酶 1(TBK1)复合体切割后，分别导入肿瘤坏死因子受体相关因子 2(TRAF2)质粒和 TANK结合激酶 1(TBK1)质粒，观察其对 NF-kB表达的影响。同时， EV71感染后，通过 RT-PCR和 Western blotting法对细胞内相关基因表达，并对 EV71 3C活性进行检测，得到蛋白酶活性与 NF-kB表达关系，判断其关联。结果 EV71感染可诱导 TBK1和其复合体蛋白 TAB 1\2\3的降解，并呈现时间依赖性( 24h:TAB1:125.86±39.87；TAB2:352.12±42.36；TAB3:195.20±29.75)，但对于肿瘤坏死因子受体相关因子 2(TRAF2)和 TANK结合激酶 1(TBK1)表达无影响。同时，细胞内白细胞介素 -6(IL-6)(19.4±0.62)、白细胞介素 -8(IL-8)(13.8±2.89)、白细胞介素 -12(IL-12)(13.0±0.68)及 IL-1β(14.6±0.96)的表达均在 12h后显著提高； TAB2可诱导 NF-kB激活，而与 3C共同表达时可显著抑制 NF-kB表达， 3C亦可显著抑制由 TAK1复合体作用诱导 NF-kB的激活。不同浓度 EV71感染后，细胞内 EV71 3C活性出现明显差异，而随着蛋白酶活性的增加，细胞内 NF-kB mRNA和蛋白的表达均成明显的下调趋势。结论肠道病毒 71型感染的星形胶质细胞中 EV71 3C可通过调节 TAK1蛋白表达，达到抑制 NF-κB信号转导的作用。 |
| 英文摘要: |
| Objective To investigate the association between the activity of enterovirus 71 type 3C protease and NF-κB signal transduction efficiency in astrocytes infected with enterovirus 71.Methods The starting and ending time of this study was from March2018 to March 2019. After infecting astrocytes with EV71 virus, the Tank-bound kinase 1 (TBK1) complex was cleaved after plasmid extraction, transformation, site-directed mutagenesis and gene transformation, and then transferred into the tumor necrosis factor receptor-associated factor 2 (TRAF2) plasmid and the TBK1 plasmid respectively to observe its effect on NF-κB expression. At the sametime, after EV71 infection, the expression of intracellular related genes and EV71 3C protease activity were detected by RT-PCR and Western blotting, and the correlation was determined according to the relationship between activity and NF-κB expression.Results EV71 infection induced the degradation of TBK1 and its complex protein tab1/2/3 in a time-dependent manner (24h: tab1: 125.86±39.87; TAB2: 352.12 ± 42.36; TAB3: 195.20 ± 75), but had no effect on the expression of TRAF2 and TBK1. At the same time, IL-6 (19. 4 4 ±0.62), IL-8 (13. 80 ±0.89), IL-12 (13. 02 ±0.68) and IL-1β(14.6±0.96) increased after 12 hours of infection; TAB2 could induce the activation of NF-κB, but co-expression with 3C could significantly inhibit the expression of NF-κB. 3C could also significantly inhibit the activation of NF-κB induced by TAK1 complex. However, with the increase of protease activity, the expression of NF-κB mRNA and protein decreased significantly.Conclusion Enterovirus 71 type 3C protease in astrocytes infected with enterovirus 71 can inhibit the transcription of NF-κB by regulating the expression of TAK1 protein. |
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