Immunological mechanism of antiviral effect of cepharanthine
Objective To investigate the antiviral effects of cepharanthine(Cep)in vivo and in vitro and explore its immunological mechanism based on the innate antiviral immune pathway.Methods The viability of A549 cells was assessed using the CCK-8 assay to determine the impact of Cep(0.062 5-64.000 0 μmol·L1).Using a vesicular stomatitis virus expressing green fluorescent protein(VSV-GFP)infected cell models,combined with flow cytometry to investigate the impact of Cep on virus replication.We also examined how pretreatment,the adsorption process,and post-adsorption treatment affect virus replication..Real-time fluorescence quantitative PCR(qRT-PCR)was employed to detect the replication of influenza A virus(H1N1),encephalomyocarditis virus(EMCV),and herpes simplex virus type 1(HSV-1)under the influence of Cep.The in vivo antiviral effects of Cep were investigated using the VSV infection mouse model.Bioinformatics analysis was conducted in A549 cells to explore the mechanism of the antiviral function of Cep.qPCR was used to detect changes in mRNA expression of IFNB1 and the interferon-stimulated genes(ISGs)in A549 and MEF cells with Cep treatment.Immunoblotting was performed to measure changes in the phosphorylation level of TBK1 and STAT1 in THP-1 cells.Results In vitro experiments showed that Cep significantly inhibited the replication of VSV,H1N1,EMCV,and HSV-1 viruses compared to the model group in A549 cells.Cep did not affect the adsorption process of VSV,but pre-treatment or post-adsorption administration effectively inhibited virus replication.Cep increased the survival rate of VSV-infected mice.Cep activates the innate antiviral immune response through the IFN-Ⅰ pathway.Conclusion This study demonstrates that Cep exhibits antiviral effects both in vivo and in vitro by activating the innate antiviral immune response through the IFN-Ⅰ pathway.
NETL
NSTL
万方数据
