黄芩苷联合连翘苷对甲型流感病毒核蛋白基因表达的影响  

Effect of baicalin combined with phillyrin on the gene expression of influenza A virus nucleoprotein

作　　者：黄智生[1] 杨斌[1] 张清[1] 孙坚[2] HUANG Zhi-sheng;YANG Bin;ZHANG Qing;SUN Jian(Department of Respiration,Fourth Affiliated Hospital of Nanchang University,Nanchang 330003,Jiangxi,China;Molecular Medicine and Genetics Center,Fourth Affiliated Hospital of Nanchang University,Nanchang 330003,Jiangxi,China)

机构地区：[1]南昌大学第四附属医院呼吸科,南昌医学硕士330003 [2]南昌大学第四附属医院分子医学与基因中心实验室,南昌330003

出　　处：《医学研究生学报》2018年第10期1033-1037,共5页Journal of Medical Postgraduates

基　　金：江西省教育厅科学技术研究项目基金(GJJ13194)

摘　　要：目的病毒的高变异率,使得抗病毒药物的研发显得非常重要,文中探讨黄芩苷联合连翘苷对甲型流感病毒核蛋白(Nucleoprotein,NP)基因表达的影响。方法通过瞬时转染技术将甲型流感病毒核蛋白真核重组质粒pc DNA3.1(+)/核蛋白导入Hela细胞以建立甲型流感病毒核蛋白真核表达载体,检测黄芩苷(15.625μg/m L)联合连翘苷(12.5μg/m L)组核蛋白基因表达量。选用低、中浓度的黄芩苷(IC25、IC50)联合低、中、高浓度的连翘苷(IC25、IC50、IC75)进行药物联合实验,实验分为Hela细胞对照组;脂质体对照组;重组质粒转染组;黄芩苷IC25+连翘苷IC25组;黄芩苷IC25+连翘苷IC25组;黄芩苷IC25+连翘苷IC75组;黄芩苷IC50+连翘苷IC50组;黄芩苷IC50+连翘苷IC50组;黄芩苷IC25+连翘苷IC75组。转染后使用各实验组药物进行干预,继续培养48 h后,采用反转录-实时荧光定量PCR(RT-qPCR)测定各实验组Hela细胞内核蛋白基因的拷贝量,评价黄芩苷联合连翘苷对靶细胞核蛋白基因表达的影响,并分析药物联合作用效果。结果重组质粒转染组核蛋白基因表达量为(62.868±13.587)×104copies/μL,Hela细胞对照组为(2.131±0.172)×104copies/μL,脂质体对照组为(1.627±0.489)×104copies/μL,黄芩苷联合连翘苷组为(7.596±2.066)×104copies/μL。黄芩苷IC25+连翘苷IC25组、黄芩苷IC25+连翘苷IC50组、黄芩苷IC25+连翘苷IC75组对靶细胞核蛋白基因表达的抑制率分别为:(37.87±9.11)%、(58.65±5.53)%、(79.12±5.35)%。通过Compu Syn软件分析药物联合指数CI分别为:1.242、0.954、0.947。黄芩苷IC50+连翘苷IC25组、黄芩苷IC50+连翘苷IC50组、黄芩苷IC50+连翘苷IC75组对靶细胞核蛋白基因表达的抑制率分别为:(62.00±4.26)%、(70.87±3.06)%、(83.00±2.90)%。通过Compu Syn软件分析药物联合指数CI分别为:0.895、0.855、0.892。黄芩苷联合连翘苷对核蛋白基因的表达的抑制作用总体表现为协同作用,低浓度黄芩苷与�Objective Influenza A(InfA)is an acute respiratory infectious disease persistently threatening human health and social stability.The high mutation of the InfA virus makes very significant the development of anti-influenza drugs.Traditional Chinese herbal drugs have shown distinct advantages in the prevention and management of InfA.This study aimed to investigate the effect of baicalin combined with phillyrin on the gene expression of InfA virus nucleoprotein(NP). Methods The nucleoprotein eukaryotic expression vector of InfA virus was constructed by transient transfection of InfA virus NP eukaryotic recombinant plasmid pcDNA3.1(+)/NP into hela cells.Six groups were designed for the experiment:hela cells,liposome,recombinant plasmid(transfected pcDNA3.1(+)/NP),baicalin,phillyrin,baicalin(15.625μg/mL)+phillyrin(12.5μg/mL),each treated with respective agents after transfection.After 48 hours of culture,the number of the copied InfA virus NPs in the hela cells was measured by RT-qPCR and the effect of baicalin combined with phillyrin on the gene expression of InfA virus NP in the target cells evaluated by the statistical method.Results RT-qPCR showed that the gene expression of InfA virus NP was significantly lower in the baicalin+phillyrin than in the recombinant plasmid group(t=6.966,P<0.01).Dose-effect analysis revealed an antagonistic effect of low-dose baicalin+low-dose phillyrin(combination index[CI]>1)and a synergistic effect of medium-and high-dose baicalin+medium-and high-dose phillyrin(CI<1). Conclusion Baicalin combined with phillyrin can decrease the gene expression of InfA virus NP and has a synergistic effect within a certain dose range.

关 键 词：黄芩苷 连翘苷 甲型流感病毒核蛋白 联合作用 基因表达 

分 类 号：R511[医药卫生—内科学]