Inhibitory Effect And Mechanism Studies Of Antivirals Against Zika Virus In Vivo And Vitro

Objective: The first part of this study is to study the in vivo and in vitro effects of emodin on zika virus(ZIKV)and its mechanism.The second part is the study of in vivo and in vitro effects of Arbidol Hydrochloride on zika virus and its mechanism.It provides theoretical and experimental basis for the follow-up research and development of new anti-zika virus drugs and clinical treatment of zika virus at the cell and animal levels.Methods:(1)The cytotoxicity of emodin on A549 cells was evaluated by MTT.Emodin was used to treat A549 cells infected by zikv in four ways: pre-treatment,biosynthesis,entry and attachment,and the expression of ZIKV NS5 gene and related antiviral gene was detected by real-time PCR.The expression of ZIKV envelope protein was detected by western blot and indirect immunofluorescence.This study also explored the pharmacodynamics of emodin in the mouse viral encephalitis model through histopathology and immunohistochemistry.(2)The cytotoxicity of arbidol hydrochloride on A549 cells was evaluated by MTT.The A549 cells infected with ZIKV were treated with arbidol in the way of prevention and biosynthesis,and the expression of ZIKV NS5 gene and related antiviral gene was detected by real-time PCR.The expression of ZIKV envelope protein was detected by western blot and indirect immunofluorescence.The pharmacodynamics of Arbidol Hydrochloride in the mouse model of viral encephalitis were investigated by histopathology and IHC.Results:(1)The CC50 of emodin on A549 cells was 31.93μM.Emodin can pre-treat ZIKV in vitro,The EC50 of emodin on A549 cells was 1.25μM,emodin can prevent zikv in vitro throught biosynthetic,the EC50 of emodin on A549 cells was 1.59μM,and the results of pre-treatment assay and biosynthetic assay showed that there are significant differences on the group of virus control and drug treatment.Emodin inhibits the expression of ZIKV NS5 gene and envelope protein(e)through pre-treatment and anti-biosynthesis.Emodin has no effect on zikv in vitro throught adsorption and invasion,and there is no statistical difference on the group of virus control and drug treatment.Emodin promoted the expression of IFN-β,IL-28,IL-29,STAT-1,STAT-2,IFITM-3,ISG-15 and APOBEC3 G in ZIKV infected A549 cells.The effect of emodin on ZIKV is related to immune responses that type Ⅰ and type Ⅲ interferon have mediated.Emodin can inhibit the replication of the virus and the pathological changes in the mouse brain,delay the onset time of the sick mice,and improve the health status and survival rate of the milk mice through treatment and pre-treatment.(2)The CC50 of ARB on A549 cells was 25.43μg/ml.Emodin can pre-treat ZIKV in vitro,The EC50 of ARB on A549 cells was 0.46μg/ml,emodin can prevent zikv in vitro throught biosynthetic,the EC50 of emodin on A549 cells was0.28μg/ml,and the results of pre-treatment assay and biosynthetic assay showed that there are significant differences on the group of virus control and drug treatment.Emodin promoted the expression of IFN-β,IL-28,IL-29,STAT-1,STAT-2,IFITM-3and APOBEC3 G in ZIKV infected A549 cells.The effect of emodin on ZIKV is related to immune responses that type Ⅰ and type Ⅲ interferon have mediated.Emodin can inhibit the replication of the virus and the pathological changes in the mouse brain,delay the onset time of the sick mice,and improve the health status and survival rate of the milk mice through treatment and pre-treatment.Conclusion:(1)Emodin can efficiently inhibit ZIKV throught pre-treatment and biosynthesis,but has no effect on zikv throught entry and attachment.Emodin plays an antiviral role by modulating cytokines through the jak-stat signaling pathway because of type Ⅰ and type Ⅲ interferon-mediated immune responses.Emodin has protective effect on mice infected with ZIKV.(2)ARB can inhibit ZIKV in vitro and vivo.ARB can inhibit the expression of ZIKV NS5 gene in vivo and improve the health status and survival rate.ARB plays an antiviral role by modulating cytokines through the jak-stat signaling pathway because of type Ⅰ and type Ⅲ interferon-mediated immune responses.