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Archive > Volume 16 Issue 1 > 2024,16(1):106-112. DOI:https://doi.org/10.1016/j.chmed.2023.09.005 Prev Next

白頭翁皂苷B4在體外和體內(nèi)抑制新冠病毒復(fù)制

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Anemoside B4 inhibits SARS-CoV-2 replication in vitro and in vivo

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    摘要:
    目的：白頭翁中含量最多的三萜皂苷——白頭翁皂苷B4（AB4）對流感病毒FM1和肺炎克雷伯菌所致肺炎均具有抑制作用。然而，AB4是否具有抗新冠病毒（SARS-CoV-2）的作用還不清楚。因此，本研究旨在研究AB4在體內(nèi)和體外抑制SARS-CoV-2的抗病毒活性及其潛在作用機(jī)制。 方法：使用CCK-8試劑盒檢測AB4的細(xì)胞毒作用；用SARS-CoV-2感染的HEK293T、HPAEpiC和Vero E6細(xì)胞進(jìn)行體外抗病毒活性研究；采用SARS-CoV-2感染的hACE2-IRES-luc轉(zhuǎn)基因小鼠模型評價(jià)AB4在體內(nèi)的抗病毒作用。此外，通過無標(biāo)記定量蛋白質(zhì)組學(xué)和生物信息學(xué)分析探討AB4潛在的抗病毒作用機(jī)制。使用免疫印跡和免疫組織化學(xué)方法檢測I型干擾素（IFN）信號通路相關(guān)蛋白的水平。 結(jié)果：AB4抑制HEK293T細(xì)胞中SARS-CoV-2的拷貝數(shù)，同時(shí)降低病毒核衣殼蛋白（N）、刺突蛋白（S）和3C樣蛋白（3CLpro）的水平；體內(nèi)抗病毒實(shí)驗(yàn)表明，在SARS-CoV-2感染小鼠模型中，AB4抑制病毒復(fù)制并抑制病毒誘導(dǎo)的肺炎。進(jìn)一步研究發(fā)現(xiàn)，AB4的抗病毒活性通過激活視黃酸誘導(dǎo)基因I（RIG-1）樣受體（RLP）途徑增強(qiáng)干擾素-β反應(yīng)相關(guān)。此外，非標(biāo)記定量蛋白質(zhì)組學(xué)分析發(fā)現(xiàn)，在SARS-CoV-2病毒感染細(xì)胞中，AB4顯著改變17種蛋白質(zhì)，這些蛋白質(zhì)主要聚集在RNA代謝中。 結(jié)論：AB4通過RLR途徑抑制SARS-CoV-2的復(fù)制，并調(diào)節(jié)RNA代謝，有望開發(fā)為抗SARS-CoV-2藥物的潛在先導(dǎo)化合物。

    Abstract:
    Objective: Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARSCoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro. Methods: The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining. Results: The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-b response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism. Conclusion: Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.

    關(guān)鍵詞:
    白頭翁皂苷B4;干擾素;RNA代謝;新型冠狀病毒

    Keywords:


    Project Supported:
    This work was jointly supported by National Natural Science Foundation of China (No. 82341087, 82073912) and Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions.

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