黄芩总黄酮对甲型流感病毒核蛋白表达的影响

摘要

目的 探讨黄芩总黄酮(Total flavonoids of Scutellaria baicalensis Georgi,TFSB)对甲型流感病毒核蛋白(NP)的干预作用.方法 本实验设HeLa细胞组、pcDNA3.1(+)空载体组、pcDNA3.1(+)、NP组、TFSB组,其中pcDNA3.1(+)空载体组、pcDNA3.1(+)/NP组分别通过瞬时转染将重组质粒pcDNA3.1(+)、pcDNA3.1(+)/NP转染到HeLa细胞中；TFSB组在将重组质粒pcDNA3.1(+)、NP转染到HeLa细胞的同时使用TFSB进行药物干预.转染48 h后,采用胶体金免疫层析试纸法测维持液中NP的表达,荧光定量RT-PCR测NP基因起始核酸量.结果 胶体金免疫层析试纸法测NP表达示:pcDNA3.1(+)/NP组、TFSB组为阳性；荧光定量RT-PCR示:pcDNA3.1(+)/NP组和TFSB组NP基因起始拷贝数分别为(8.90±2.53)×106 copies/μ1、(6.15±1.49)×106 copies/μl,采用t检验进行统计学分析:差异无统计学意义(P＞0.05).结论 黄芩总黄酮对甲型流感病毒NP基因和NP的表达没有影响.%Objective To investigate the effect of total flavonoids of Scutellaria baicalensis Georgi (TFSB) on exogenous expression of influenza A virus nudeoprotein (NP) in HeLa cells. Methods HeLa cells were transiently transfected with the empty vector pcDNA3.1(+) or pcDNA3.1(+)/NP vector harboring influenza A virus NP. The pcDNA3.1(+)/NP-transfected cells were treated with TSFB and the expression of influenza A virus NP in the cell supernatant was measured using colloidal gold immunochromatography 48 h after the transfection; fluorescent quantitative RT-PCR was performed to measure the starting copy number of NP gene. Results The cells transfected with pcDNA3.1 (+)/NP with and without TFSB treatment were positive for NP expression. Fluorescent quantitative RT-PCR showed that the starting copy number of NP gene in pcDNA3.1(+)/ NP-transfected cells was (8.90±2.53)×106 copies/μl, showing no significant difference from that of (6.15±1.49)×106 copies/μl in pcDNA3.1(+)/NP-transfected cells with subsequent TFSB treatment (P＞0.05). Conclusion TFSB treatment does not obviously affect exogenous influenza A virus NP gene expression or its protein synthesis in HeLa cells.