论文部分内容阅读
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。
AIM: To investigate the effect of high dose of Astragalus polysaccharides on the expression of plasmid pNS1 / EGFP in the tissue of influenza A / PR / 8/34 NS1 gene. Methods: Plasmid pNS1 / EGFP containing A / PR / 8/34 NS1 was constructed. Sixty Kunming mice were randomly divided into three groups with 20 mice in each group. The mice were injected intramuscularly with p EGFP-N1, pNS1 / EGFP and pNS1 / EGFP + respectively. Muscle and serum samples were taken on the 14th and 28th days after the last injection. Serum levels of IL-4 and IFN-γ were measured by ELISA. The expression of NS1 protein and Caspase-3 in muscle was measured by Western blot. Results: At the 14th day after the last injection, the levels of IL-4 in each group were not significantly different, while the levels of IFN-γ in pNS1 / EGFP + APS group were significantly different from those in the other groups. However, the levels of caspase-3 The results of RT-PCR showed that there was no significant difference between pNS1 / EGFP + APS group and pNS1 / EGFP. The levels of IFN-γ and IL-4 in pNS1 / EGFP + astragalus polysaccharides group were significantly different from those in the other groups on day 28, while the expression of caspase-3 in pNS1 / EGFP + astragalus polysaccharide group was significantly different from that in pNS1 / EGFP group The expression level of NS1 in pNS1 / EGFP + APS group was also significantly different from that in pNS1 / EGFP group. Conclusion: High dose of astragalus polysaccharide can prolong the expression of pNS1 / EGFP, and this effect is related to the reduction of inflammation and the decrease of apoptosis.