连翘苷对甲型流感病毒核蛋白基因表达的影响研究

目的探讨连翘苷对甲型流感病毒核蛋白(NP)基因转染后表达的影响。方法将甲型流感病毒NP基因转染Hela细胞,用甲型流感病毒胶体金法检测连翘苷对转染后细胞内和上清核蛋白表达情况,用实时定量反转录聚合酶链反应(RT-PCR)检测Hela细胞内NP基因的拷贝数。结果 NP重组质粒组甲型流感病毒核蛋白含量高。空质粒组、脂质体组、Hela细胞组基本不含甲型流感病毒核蛋白。连翘苷组上清无或可能含有微量核蛋白。连翘苷组胞内甲型流感病毒含量不高。RT-PCR校正曲线相关性为0.998,效率为97.4%。重复4次转染后48 h连翘苷组NP基因表达量为(2.1±0.3)×105拷贝数/μl,NP重组质粒组NP基因表达量为(61.5±15.0)×105拷贝数/μl,连翘苷组NP基因表达量低于NP重组质粒组,差异有统计学意义(t=7.672,P<0.05)。结论连翘苷可以抑制甲型流感病毒NP基因转染后表达。

Effect of Phillyrin on Gene Expression of Influenza A Virus Nucleoprotein

Objective To investigate the effect of phillyrin on gene expression of influenza A virus nucleoprotein after transfection.Methods Hela cells were transfected with NP gene of influenza A virus,and Influenza A virus colloidal gold method was used to detect the phillyrin supernatant and intracellular nucleoprotein expression,and RT-PCR was used to detect the NP gene copy number.Results NP recombinant plasmid group contained high level influenza A virus nucleoprotein.Empty plasmid group,liposome group and Hela cells group mainly did not contain influenza A virus nucleoprotein.Phillyrin supernatant contained no or little influenza A virus nucleoprotein.Phillyrin contained low level intracellular influenza A virus nucleoprotein.RT-PCR calibration curve showed the correlation was 0.998 and the efficiency was 97.4%.48 h after four times transfection,NP gene expression level was(2.05±0.3)×105 copy number/μl in Phillyrin group,(61.5±15.0)×105 copy number/μl in recombinant plasmid group,significantly higher than Phillyrin group.The difference of NP gene expression level between Phillyrin group and recombinant plasmid group was statistically significant(t=7.672,P<0.O5).Conclusion Phillyrin can inhibit the expression of Influenza A virus NP gene after transfection.