中华临床感染病杂志
中華臨床感染病雜誌
중화림상감염병잡지
CHINESE JOURNAL OF CLINICAL INFECTIOUS DISEASES
2015年
2期
133-138
,共6页
彭雁忠%张仁利%郑立运%张崇远%刘利成
彭雁忠%張仁利%鄭立運%張崇遠%劉利成
팽안충%장인리%정립운%장숭원%류리성
干扰素α-2b%奥司他韦%流感病毒A型%流感病毒B型%体外研究
榦擾素α-2b%奧司他韋%流感病毒A型%流感病毒B型%體外研究
간우소α-2b%오사타위%류감병독A형%류감병독B형%체외연구
Interferon α-2b%Oseltamivir%Influenza A virus%Influenza B virus%In vitro study
目的 研究重组人干扰素(IFN) α-2b对流感病毒的体外抑制作用.方法 将甲型H1N1流感病毒和乙型流感B/Y病毒接种到体外Vero细胞中,并添加不同浓度的IFNα-2b和奥司他韦共培养.观察并计算病毒空斑数,同时利用荧光定量RT-PCR法比较IFNα-2b和奥司他韦对流感病毒的体外抑制效果,利用荧光显微镜观察病毒核糖核蛋白(RNP)复合物的输出.结果 病毒空斑试验显示,10 μg/μL的IFNα-2b和10 μg/μL奥司他韦对甲型H1N1流感病毒具有明显的抑制作用,病毒空斑数分别为7.5×108和15×108PFU/mL,其中,奥司他韦对病毒的抑制作用较IFNα-2b更强;同样,10 μg/μL IFNα-2b和10μg/μL奥司他韦对乙型流感B/Y病毒均具有明显的抑制作用,病毒空斑形成单位分别为1.1×108和1.5×108PFU/mL.荧光定量RT-PCR检测结果显示,加入10 μmol/LIFNα-2b和10 μmol/L奥司他韦的培养基中甲型H1N1流感病毒和乙型流感B/Y病毒核酸扩增循环阈值(CT)均明显高于流感病毒单独培养组,其中,甲型H1N1流感病毒单独培养组、IFNα-2b和奥司他韦共培养组的病毒核酸CT值分别为16、26和35,乙型流感B/Y病毒单独培养组、IFNα-2b和奥司他韦共培养组的病毒核酸CT值分别为18、27和31.荧光显微镜观察病毒RNP复合物的输出发现,10 μmol/L IFNα-2b能够有效损害感染Vero细胞内病毒RNP复合物的输出.结论 IFNα-2b在体外对甲型H1N1流感病毒和乙型流感B/Y病毒均具有明显的抑制作用.
目的 研究重組人榦擾素(IFN) α-2b對流感病毒的體外抑製作用.方法 將甲型H1N1流感病毒和乙型流感B/Y病毒接種到體外Vero細胞中,併添加不同濃度的IFNα-2b和奧司他韋共培養.觀察併計算病毒空斑數,同時利用熒光定量RT-PCR法比較IFNα-2b和奧司他韋對流感病毒的體外抑製效果,利用熒光顯微鏡觀察病毒覈糖覈蛋白(RNP)複閤物的輸齣.結果 病毒空斑試驗顯示,10 μg/μL的IFNα-2b和10 μg/μL奧司他韋對甲型H1N1流感病毒具有明顯的抑製作用,病毒空斑數分彆為7.5×108和15×108PFU/mL,其中,奧司他韋對病毒的抑製作用較IFNα-2b更彊;同樣,10 μg/μL IFNα-2b和10μg/μL奧司他韋對乙型流感B/Y病毒均具有明顯的抑製作用,病毒空斑形成單位分彆為1.1×108和1.5×108PFU/mL.熒光定量RT-PCR檢測結果顯示,加入10 μmol/LIFNα-2b和10 μmol/L奧司他韋的培養基中甲型H1N1流感病毒和乙型流感B/Y病毒覈痠擴增循環閾值(CT)均明顯高于流感病毒單獨培養組,其中,甲型H1N1流感病毒單獨培養組、IFNα-2b和奧司他韋共培養組的病毒覈痠CT值分彆為16、26和35,乙型流感B/Y病毒單獨培養組、IFNα-2b和奧司他韋共培養組的病毒覈痠CT值分彆為18、27和31.熒光顯微鏡觀察病毒RNP複閤物的輸齣髮現,10 μmol/L IFNα-2b能夠有效損害感染Vero細胞內病毒RNP複閤物的輸齣.結論 IFNα-2b在體外對甲型H1N1流感病毒和乙型流感B/Y病毒均具有明顯的抑製作用.
목적 연구중조인간우소(IFN) α-2b대류감병독적체외억제작용.방법 장갑형H1N1류감병독화을형류감B/Y병독접충도체외Vero세포중,병첨가불동농도적IFNα-2b화오사타위공배양.관찰병계산병독공반수,동시이용형광정량RT-PCR법비교IFNα-2b화오사타위대류감병독적체외억제효과,이용형광현미경관찰병독핵당핵단백(RNP)복합물적수출.결과 병독공반시험현시,10 μg/μL적IFNα-2b화10 μg/μL오사타위대갑형H1N1류감병독구유명현적억제작용,병독공반수분별위7.5×108화15×108PFU/mL,기중,오사타위대병독적억제작용교IFNα-2b경강;동양,10 μg/μL IFNα-2b화10μg/μL오사타위대을형류감B/Y병독균구유명현적억제작용,병독공반형성단위분별위1.1×108화1.5×108PFU/mL.형광정량RT-PCR검측결과현시,가입10 μmol/LIFNα-2b화10 μmol/L오사타위적배양기중갑형H1N1류감병독화을형류감B/Y병독핵산확증순배역치(CT)균명현고우류감병독단독배양조,기중,갑형H1N1류감병독단독배양조、IFNα-2b화오사타위공배양조적병독핵산CT치분별위16、26화35,을형류감B/Y병독단독배양조、IFNα-2b화오사타위공배양조적병독핵산CT치분별위18、27화31.형광현미경관찰병독RNP복합물적수출발현,10 μmol/L IFNα-2b능구유효손해감염Vero세포내병독RNP복합물적수출.결론 IFNα-2b재체외대갑형H1N1류감병독화을형류감B/Y병독균구유명현적억제작용.
Objective To investigate the effect of recombinant human interferon α-2b on influenza virus in vitro.Methods Influenza A virus subtype H1N1 and influenza B/Y virus were inoculated into Vero cells and different concentrations of interferon α-2b and oseltamivir were added.Numbers of virus plaques were observed and calculated,and quantitative RT-PCR were used to assess the inhibitory effect of interferon α-2b and oseltamivir in vitro.The nuclear export of viral ribonucleoprotein (RNP) complexes were monitored under fluorescence microscope.Results Virus plaque test showed that influenza A viruses subtype H1N1 were significantly inhibited when 10 μg/μL interferon α-2b and 10 μg/μL oseltamivir were added,and the numbers of plaques were 7.5 × 108 and 15 × 108 PFU/mL,respectively;the inhibitory effect of oseltamivir was better than that of interferon α-2b.Influenza B/Y viruses were also inhibited when 10 μg/μL interferon α-2b and 10 μg/μL oseltamivir were added,and the numbers of plaques were 1.1 × 108 and 1.5 × 108 PFU/mL,respectively.Quantitative RT-PCR results showed that the cycle threshold (CT) values of influenza A virus subtype H1N1 and influenza B/Y virus were much higher when 10 μmol/L interferon α-2b and 10 μmol/L oseltamivir were added.CT values of influenza A virus subtype H1N1 were 16,26 and 35 before and after inferferon α-2b and oseltamivir were added.CT values of influenza B/Y virus were 18,27 and 31 before and after interferon α-2b and oseltamivir were added.Reduction in the nuclear export of viral RNP in influenza A virus subtype H1N1-infected Vero cells was also observed when 10 μmol/L interferon α-2b were added.Conclusion Interferon α-2b has significantly inhibitory effect on both influenza A virus subtype H1N1 and influenza B/Y virus in vitro.
