MODE OF ACTION OF THE ANTIINFLUENZA VIRUS ACTIVITY OF PLANT FLAVONOID, 5,7,4'-TRIHYDROXY-8-METHOXYFLAVONE, FROM THE ROOTS OF SCUTELLARIA-BAICALENSIS
MODE OF ACTION OF THE ANTIINFLUENZA VIRUS ACTIVITY OF PLANT FLAVONOID, 5,7,4'-TRIHYDROXY-8-METHOXYFLAVONE, FROM THE ROOTS OF SCUTELLARIA-BAICALENSIS
复制标题
DOI：
10.1016/0166-3542(94)00062-d
复制
发表时间：
1995-01-01
影响因子：
7.6
通讯作者：
YAMADA, H
中科院分区：
文献类型：
--
作者：
NAGAI, T;MORIGUCHI, R;YAMADA, H
当小鼠适应的流感病毒A/PR/8/34（A/PR 8）（10 PFU/细胞）在4 ℃下吸附至Madin-Darby犬肾（MDCK）细胞1小时并在37 ℃下孵育时，从孵育后4小时起在培养基中检测到病毒从细胞的释放，并在8小时达到平台期。而黄芩根中的5，7，4 ′-三羟基-8-甲氧基黄酮（F36）在孵育后4 h至12 h以剂量依赖的方式显著抑制A/PR 8的这种单循环复制，使病毒滴度降低十分之一的剂量为11 μ M。F36（50 μ M）不抑制A/PR 8对MDCK细胞的吸附，但在孵育后0或2小时加入时减少培养基中病毒的释放。通过唾液酸酶活性测定的细胞相关病毒也在0或2 h时被F36处理减少。在pH5.0时，F36还抑制A/PR 8与含有牛脑混合神经节苷脂的脂质体的融合。而F36对病毒RNA依赖的RNA聚合酶的体外延伸活性影响不大。这些结果表明，F36通过抑制病毒与内体/溶酶体膜的融合来减少A/PR 8的复制，该融合发生在病毒感染周期的早期阶段。而当F36加入到A/PR 8感染的MDCK细胞中时，在孵育后3或4小时，与对照相比，培养基中病毒的释放减少，但细胞相关病毒增加。扫描和透射免疫电镜研究表明，F36抑制后代A/PR 8从MDCK细胞表面和微绒毛的出芽，当它在孵育后3小时加入。通过加入F36，通过免疫荧光和透射免疫电镜观察到A/PR 8抗原在细胞表面上的积累。这些结果表明，当在病毒感染周期的出芽阶段加入F36时，F36还通过抑制子代病毒从细胞表面出芽而显示出针对A/PR 8的抗流感病毒活性。
When mouse-adapted influenza virus A/PR/8/34 (A/PR8) (10 PFU/cell) was adsorbed to Madin-Darby canine kidney (MDCK) cells at 4 degrees C for 1 h and incubated at 37 degrees C, release of the virus from the cells was detected in the medium from 4 h after incubation and reached to plateau at 8 h. However, 5,7,4'-trihydroxy-8-methoxyflavone (F36) from the roots of Scutellaria baicalensis significantly reduced this single-cycle replication of A/PR8 from 4 h to 12 h after incubation by dose-dependent manner and the dose which decrease the virus titer one tenth was 11 mu M. F36 (50 mu M) did not inhibit the adsorption of A/PR8 to MDCK cells, but reduced release of the virus in the medium, when it was added at 0 or 2 h after the incubation. The cell-associated virus determined by sialidase activity was also reduced by F36 treatment at 0 or 2 h. F36 also inhibited the fusion of A/PR8 with liposomes containing bovine brain mixed gangliosides at pH 5.0. However, F36 little affected on the elongation activity of the viral RNA-dependent RNA polymerase in vitro. These results suggest that F36 reduces the replication of A/PR8 by inhibiting the fusion of the virus with endosome/lysosome membrane which occurs at early stage of virus infection cycle. Whereas, when F36 was added to the MDCK cells infected with A/PR8 at 3 or 4 h after incubation, release of the virus in the medium was reduced but the cell-associated virus was increased in comparison with control. Scanning and transmission immunoelectron microscopic studies revealed that F36 inhibited the budding of progeny A/PR8 from the MDCK cell surface and microvilli, when it was added at 3 h after incubation. The accumulation of the A/PR8 antigen was observed on the cell surface by immunofluorescence and transmission immunoelectron microscopies by the addition of F36. These results suggest that F36 also shows anti-influenza virus activity against A/PR8 by inhibiting the budding of the progeny virus from the cell surface, when it was added at budding stage of virus infection cycle.