连翘苷对甲型流感病毒核蛋白基因表达的影响研究  

Effect of Phillyrin on Gene Expression of Influenza A Virus Nucleoprotein

作　　者：段林建[1] 张清[1] 王农荣[2] 杨斌[2] 何士勤[2] 孙坚[2] 

机构地区：[1]南昌大学医学院,江西省南昌市330006 [2]南昌大学第四附属医院

出　　处：《中国全科医学》2012年第18期2082-2084,共3页Chinese General Practice

基　　金：国家自然科学基金项目(30860351);江西省卫生厅中医药科研基金(2008ZZ0014)

摘　　要：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。Objective To investigate the effect of phillyrin on gene expression of influenza A virus nueleoprotein after transfeetion. Methods Hela cells were transfected with N P gene of influenza A virus, and Influenza A virus colloidal gold meth- od was used to detect the phillyrin supernatant and intraeellular 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 plas- mid group, liposome group and Hela cells group mainly did not contain influenza A virus nucleopratein. Phillyrin superuatant con- tained no or little influenza A virus nucleoprotein. Phillyrin contained low level intraeellular influenza A virus nueleoprotein. RT - PCR calibration curve showed the correlation was 0. 998 and the efficiency was 97.4%. 48 h after four times transfeetion, 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. 05 ） . Conchtsion Phillyrin can inhibit the expression of Influenza A virus NP gene after transfection.

关 键 词：连翘苷 甲型流感病毒 NP基因 

分 类 号：R285[医药卫生—中药学]