三种板蓝根制剂对流感病毒核蛋白表达的影响  

Effect of three kinds of radix isatidis preparation on the expression of nucleoprotein of influenza virus

作　　者：徐咏书[1] 孙坚[2] 何士勤[1] 

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

出　　处：《山东医药》2010年第27期8-10,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30860351)

摘　　要：目的探讨不同板蓝根制剂对甲型流感病毒核蛋白(NP)的作用,寻找抗流感病毒中药的有效成分。方法采用传统水煮浸法提取的板蓝根水煎液和有机溶剂浸泡法提取的板蓝根凝集素及购买的板蓝根注射液,用细胞病变效应(CPE)法测得三种制剂试验浓度分别为1.95、2.60、0.78 mg/ml。将培养的Hela细胞分为八组,质粒组转染pcDNA3.1(+)/NP,空质粒组转染pcDNA3.1(+);注射液、水煎液、凝集素实验组则将板蓝根注射液、水煎液、凝集素的试验浓度分别作用于转染重组质粒pcDNA3.1(+)/NP的Hela细胞,三种制剂对应的对照组不进行转染,仅将对应制剂作用于Hela细胞。用胶体金免疫层析法和间接免疫荧光法检测各组NP表达。结果质粒组、注射液及水煎液实验组胶体金试验阳性,免疫荧光法证实作用转染的Hela细胞仍发出较强绿色荧光,且表达的NP定位于细胞质中;而凝集素组及空质粒组中表达NP的细胞未出现较强的绿色荧光且胶体金试验阴性。结论板蓝根凝集素可抑制NP基因表达,此为研发抗流感病毒的药物提供了新思路。Objective To investigate the different preparations of radix isatidis on the nucleoprotein（NP） of influenza virus,so as to look for the active ingredient of anti-influenza A virus herbs.Methods The radix isatidis decoction and isatis agglutinin were separately decocted and concentrated by boiled leaching and organic solvent immersion method,and radix isatidis injection was purchased,the test concentrations of which were 1.95,2.60,0.78 mg/ml detected by means of cytopathic effects（CPE）.The cultured Hela cells were divided into 8 groups,the plasmid group was transfected with pcDNA3.1（＋）/NP,the empty plasmid group was transfected with pcDNA3.1（＋）;after transfected with pcDNA3.1（＋）/NP,the experimental groups of injection,decoctionand,agglutinin were treated with the three preparations of radix isatidis at above test concentrations,the control groups corresponding the three preparations were not transfected and treated with the preparations only.Indirect immunofluorescence and colloidal gold immunochromatographic strip assay were used for determining the expression of NP gene.Results In the plasmid group and the decoction group and injection group of the radix isatidis,colloidal gold immunochromatography showed positive results,and strong green fluorescent was appeared in transfected Hela cells,and the NP were expressed in cytoplasm;while in the agglutinin group and the empty plasmid group,no strong green fluorescent were found in the cells expressed NP and,colloidal gold immunochromatography showed negative results.Conclusion Radix isatis agglutinin can inhibit the expression of NP gene,which provide new thoughts for the research of drugs anti-influenza virus.

关 键 词：流感病毒 板蓝根 核蛋白 转染 

分 类 号：R511.7[医药卫生—内科学]