Anemoside B4 inhibits enterovirus 71 propagation in mice through upregulating 14-3-3 expression and type I interferon responses
Anemoside B4 inhibits enterovirus 71 propagation in mice through upregulating 14-3-3 expression and type I interferon responses
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白头翁苷 B4 通过上调 14-3-3 表达和 I 型干扰素反应抑制小鼠肠道病毒 71 传播
DOI：
10.1038/s41401-021-00733-1
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发表时间：
2021-07-28
影响因子：
8.2
通讯作者：
Yu, Di
中科院分区：
文献类型：
--
作者：
Kang, Nai-xin;Zou, Yue;Yu, Di
肠病毒71型（EV71）是人类手足口病的主要病原体。EV71能有效地逃避宿主的先天免疫反应而引起感染。目前，尚无有效的EV71病毒抗病毒药物。Anemoside B4 （B4）是从白头翁（pulsatila chinensis）根中分离得到的一种天然皂苷。显示出广泛的生物活性。在本研究中，我们研究了B4在细胞培养和哺乳小鼠中对EV71病毒的抗病毒活性。我们发现B4 （12.5-200 μ M）剂量依赖性地提高了EV71感染的RD细胞的活力，其对EV71的IC50值为24.95 +/- 0.05 μ M。B4的抗病毒活性与干扰素(IFN)- β反应增强有关，因为IFN- β的敲低会消除其抗病毒活性。我们还证实了IFN反应的增强是通过激活视黄酸诱导基因I （RIG-I）样受体（RLRs）途径介导的，并通过14-3-3蛋白的上调来实现，这破坏了yes相关蛋白（YAP）和干扰素调节因子3 （IRF3）之间的相互作用。通过基于细胞培养氨基酸（SILAC）的蛋白质组学分析，我们发现Hippo通路是b4处理的ev71感染细胞中最重要的功能簇。用感染EV71病毒的哺乳小鼠（2日龄）进行体内实验，随后给予B4 (200 mg中心点kg（-1）中心点d(-1), ig) 16 d。我们发现B4有效抑制了EV71的复制，改善了肌肉炎症和肢体活动。同时，B4可调节手足口病生物标志物IL-10和ifn - γ的表达，减轻EV71感染的并发症。总之，我们的研究结果表明B4可以通过协调Hippo和RLRs通路来增强ifn - β的抗病毒作用，B4可能是开发抗ev71药物的潜在先导化合物。
Enterovirus 71 (EV71) is the major pathogens of human hand, foot, and mouth disease (HFMD). EV71 efficiently escapes innate immunity responses of the host to cause infection. At present, no effective antiviral drugs for EV71 are available. Anemoside B4 (B4) is a natural saponin isolated from the roots of Pulsatilla chinensis (Bunge) Regel. P. chinensis extracts that shows a wide variety of biological activities. In this study, we investigated the antiviral activities of B4 against EV71 both in cell culture and in suckling mice. We showed that B4 (12.5-200 mu M) dose dependently increased the viability of EV71-infected RD cells with an IC50 value of 24.95 +/- 0.05 mu M against EV71. The antiviral activity of B4 was associated with enhanced interferon (IFN)-beta response, since knockdown of IFN-beta abolished its antiviral activity. We also confirmed that the enhanced IFN response was mediated via activation of retinoic acid-inducible gene I (RIG-I) like receptors (RLRs) pathway, and it was executed by upregulation of 14-3-3 protein, which disrupted the interaction between yes-associated protein (YAP) and interferon regulatory factor 3 (IRF3). By using amino acids in cell culture (SILAC)-based proteomics profiling, we identified the Hippo pathway as the top-ranking functional cluster in B4-treated EV71-infected cells. In vivo experiments were conducted in suckling mice (2-day-old) infected with EV71 and subsequently B4 (200 mg center dot kg(-1) center dot d(-1), i.p.) was administered for 16 days. We showed that B4 administration effectively suppressed EV71 replication and improved muscle inflammation and limb activity. Meanwhile, B4 administration regulated the expressions of HFMD biomarkers IL-10 and IFN-gamma, attenuating complications of EV71 infection. Collectively, our results suggest that B4 could enhance the antiviral effect of IFN-beta by orchestrating Hippo and RLRs pathway, and B4 would be a potential lead compound for developing an anti-EV71 drug.