大剂量黄芪多糖增强甲型流感病毒NS蛋白表达的研究  

Research of increasing influenza A virus NS1 protein expression by high dose astragalus polysaccharides

作　　者：褚兆苹[1] 吴淑慧[2] 刘文泰[2] 马志红[2] 徐丙元[2] 曹刚[2] 徐华洲[2] 石玉娥[2] 戴军[2] 

机构地区：[1]河北省人民医院妇产科,石家庄050051 [2]河北中医学院基础医学院,石家庄050200

出　　处：《中国免疫学杂志》2015年第5期629-631,637,共4页Chinese Journal of Immunology

基　　金：河北省卫生厅课题(20130139)

摘　　要：目的:研究大剂量的黄芪多糖对构建有甲型流感A/PR/8/34株NS1基因的质粒pNS1/EGFP的在组织内表达的影响。方法:构建含甲流A/PR/8/34 NS1的质粒pNS1/EGFP。将60只昆明小鼠随机分为三组,每组20只,分别肌肉注射p EGFP-N1,pNS1/EGFP和pNS1/EGFP+腹腔注射黄芪多糖,间隔三周注射2次。最后一次注射后14 d和28 d分别取肌肉和血清标本。ELISA法测量血清IL-4和IFN-γ水平;Western blot法测量肌肉中NS1蛋白和Caspase-3的表达水平。结果:在最后一次注射后第14天,各组IL-4水平差别不大,而pNS1/EGFP+黄芪多糖组的IFN-γ水平与其他组有显著性差异,但caspase-3表达水平各组无显著性差异,而RT-PCR结果显示pNS1/EGFP+黄芪多糖组NS1蛋白水平与pNS1/EGFP无显著性差异。在第28天pNS1/EGFP+黄芪多糖组的IFN-γ水平和IL-4水平均与其他组有显著性差异,pNS1/EGFP+黄芪多糖组caspase-3表达与pNS1/EGFP组有显著性差异,而pNS1/EGFP+黄芪多糖组NS1的表达水平也与pNS1/EGFP组有显著性差异。结论:大剂量的黄芪多糖能够延长pNS1/EGFP表达,这种作用与炎症减轻和凋亡减少有关。Objective：To study whether high dose astragalus polysaccharides （APS） could affect the expression of pNSI/EGFP that included influenza A virus（IAV） non-structure protein 1 （NS1） gene in the tissue. Methods： pNS1/EGFP was constructured with NS1 of IAV. Sixty Kunming mice were divided into three groups randomly. Each group of mice was injected separately with one of the following：pEGFP-Nl, pNS1/EGFP and pNS1/EGFP＋ APS in intraperitoneal injection. The mice were injected by intramuscular injection twice with a 3-week interval between injections. The serum samples and muscle samples were obtained on day 14 and day 28 after the booster injection. Sera IL-4, sera IFN-7, muscle caspase-3 and muscle NS1 expression were measured in ELISA, Western blot or RT-PCR. Results ： There were no significant difference among the different groups in day 14 expect that IFN-γ of pNS1/EGFP＋APS were lower（P〈0. 05）. IFN-3, level or IL4 level of pNS1/EGFP＋APS were lower compared with other groups in day 28. caspase-3 of pNS1/EGFP＋APS were lower compared with other groups in day 28. Conclusion： APS could increase the expression of pNS1/EGFP by decreasing the inflammation and apoptosis.

关 键 词：黄芪多糖 NS1 CASPASE-3 凋亡 

分 类 号：R373.1[医药卫生—病原生物学]