论文部分内容阅读
【目的】在细胞水平上研究黄酮类化合物flavopiridol的抗流感病毒效果,初步探索了其抗流感病毒的机制。【方法】首先用Western blot初步检测了在蛋白激酶抑制剂flavopiridol处理下流感病毒NP和M1蛋白的水平,然后通过免疫荧光实验观察了宿主细胞中流感病毒vRNP的合成,又利用噬斑实验检测了flavopiridol对病毒复制的影响,最后通过检测flavopiridol处理的宿主细胞内RNA聚合酶Ⅱ的磷酸化状态和病毒各种RNA的合成量,探究了flavopiridol抑制流感病毒复制的机理。【结果】结果表明,flavopiridol在细胞水平上可以显著抑制流感病毒蛋白质和vRNP的合成及病毒的复制,同时flavopiridol也可以抑制宿主RNA聚合酶Ⅱ大亚基CTD结构域七肽重复序列中的2位丝氨酸的磷酸化来抑制聚合酶的转录延伸活性,显著地减少病毒vRNA的合成。【结论】Flavopiridol可以通过抑制宿主细胞RNA聚合酶Ⅱ的转录延伸活性有效地抑制流感病毒的复制。
【Objective】 The anti-influenza virus effect of flavopiridol was studied at the cellular level, and its anti-influenza virus mechanism was initially explored. 【Method】 The levels of NP and Ml protein of influenza virus were first detected by Western blot and then the synthesis of influenza virus vRNP in host cells was observed by immunofluorescence assay. The plaque assay was also used to detect flavopiridol on virus replication. Finally, the mechanism by which flavopiridol inhibits the replication of influenza virus was explored by examining the phosphorylation state of RNA polymerase II in host cells treated with flavopiridol and the synthesis of various RNAs of the virus. 【Result】 The results showed that flavopiridol could significantly inhibit the synthesis of influenza virus protein and vRNP and the replication of virus at the cellular level. At the same time, flavopiridol can also inhibit the 2 position in the heptad repeat sequence of the CTD domain of the large RNA subunit Ⅱ Serine phosphorylation to inhibit polymerase transcription elongation activity, significantly reducing the viral vRNA synthesis. 【Conclusion】 Flavopiridol can effectively inhibit the replication of influenza virus by inhibiting the transcriptional elongation activity of RNA polymerase Ⅱ in host cells.