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 μM)dose dependently increased the viability of EV71-infected RD cells with an IC50 value of 24.95±0.05 μM against EV71.The antiviral activity of B4 was associated with enhanced interferon(IFN)-β response,since knockdown of IFN-β 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·kg-1·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-y,attenuating complications of EV71 infection.Collectively,our results suggest that B4 could enhance the antiviral effect of IFN-β by orchestrating Hippo and RLRs pathway,and B4 would be a potential lead compound for developing an anti-EV71 drug.

Nai-xin Kang;Yue Zou;Qing-hua Liang;Yan-er Wang;Yan-li Liu;Guo-qiang Xu;Han-dong Fan;Qiong-ming Xu;Shi-lin Yang;Di Yu

College of Pharmaceutical Science,Soochow University,Suzhou 215123,China;State Key Laboratory of Bioactive Substance and Function of Natural Medicines,Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China;Institute of Aging Research,School of Medicine,Hangzhou Normal University,Hangzhou 310036,China;Department of Immunology,Genetics and Pathology,Science for Life Laboratory,Uppsala University,Uppsala,Sweden

中国药理学报（英文版）

[1]Nai-xin Kang;Yue Zou;Qing-hua Liang;Yan-er Wang;Yan-li Liu;Guo-qiang Xu;Han-dong Fan;Qiong-ming Xu;Shi-lin Yang;Di Yu-.Anemoside B4 inhibits enterovirus 71 propagation in mice through upregulating 14-3-3 expression and type Ⅰ interferon responses)[J].中国药理学报（英文版）,2022(04):977-991

Anemoside,Pulsatilla

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