This study aims to establish the culture method of human primary airway epithelial cell(hAEC)and to investigate the anti-respiratory syncytial virus(RSV)activity and mechanism of 3-thioindole compound RSVA-4 and immunosuppressive metabolite 6-MM-PR using hAEC system,which intends to construct a cell model for RSV drug screening and efficacy evaluation.The respiratory tract epithelial cells from volunteers were collected and cultured,then the morphology,activity and purity were identified.The anti-RSV ac-tivity and cytotoxicity of RSVA-4 and 6-MMPR were further verified in hAEC system.The mechanism of RSVA-4 and 6-MMPR ac-counting for the suppression of RSV replication on hAEC was explored by using fluorescence real-time quantitative PCR(RT-qPCR)and time-of-addition assay.The survival rate of cultured hAEC was 93.51％as determined by trypan blue staining.The half maximal inhibitory concentrations(IC50)of RSV-A-4 and 6-MMPR were(207.30±4.77)µmol/L and(3 191.00±6.11)μmol/L,respective-ly.The half maximal cytotoxic concentration(CC50)of 6-MMPR was(95 526.00±10.97)µmol/L,while no toxicity of RSVA-4 was observed on hAEC.Mechanistically,RSV-A-4 and 6-MMPr inhibited RSV replication in the genome replication/transcription phase.The hAEC culture method was successfully established,which could be used to screen and evaluate the anti-RSV drugs in vitro.RS-VA-4 and 6-MMPR could effectively inhibit RSV replication at the cellular level.Altogether,the result could provide an experimental basis for the research and development of RSV drug and pathogenesis.
关键词
人原代呼吸道上皮细胞/呼吸道合胞病毒/抗病毒化合物/抗病毒活性/抗病毒机制
Key words
human airway epithelial cell/human respiratory syncytial virus/antiviral compound/antiviral activity/antiviral mechanism
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