18β-glycyrrhetinic acid inhibits rotavirus replication in culture.
18β-glycyrrhetinic acid inhibits rotavirus replication in culture.
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DOI：
10.1186/1743-422x-9-96
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发表时间：
2012-05-22
期刊：
影响因子：
4.8
通讯作者：
Faunce NR
中科院分区：
文献类型：
--
作者：
Hardy ME;Hendricks JM;Paulson JM;Faunce NR
甘草甜素 (GA) 和主要代谢物 18β-甘草次酸 (GRA) 是药用甘草根的药理活性成分，两者均已被证明具有抗病毒和免疫调节特性。尽管这些特性已得到充分证实，但其作用机制尚未完全了解。在这项研究中，测试了 GA 和 GRA 在细胞培养物中抑制轮状病毒复制的能力，以实现发现可补充现有疫苗的天然化合物的长期目标。在感染前或感染后用 GA 或 GRA 处理上皮细胞不同时间，并通过免疫荧光焦点测定法测量病毒产量。通过免疫印迹测量 GRA 处理的细胞中病毒蛋白 VP2、VP6 和 NSP2 的水平，以确定 GRA 处理是否对病毒蛋白的积累有影响。当病毒吸附后添加到受感染的培养物中时，GRA 处理使轮状病毒产量降低了 99%，而 GA 处理的培养物中的病毒产量与模拟处理的对照相似。添加时间实验表明GRA介导的复制抑制可能发生在病毒进入后的一个或多个步骤中。当在进入后两小时内将 GRA 添加到培养物中时，VP2、VP6 和 NSP2 的量显着减少。 GRA（而非 GA）在体外对轮状病毒复制具有显着的抗病毒活性，确定 GRA 是否能减弱体内轮状病毒复制的研究正在进行中。
Glycyrrhizin (GA) and primary metabolite 18β-glycyrrhetinic acid (GRA) are pharmacologically active components of the medicinal licorice root, and both have been shown to have antiviral and immunomodulatory properties. Although these properties are well established, the mechanisms of action are not completely understood. In this study, GA and GRA were tested for the ability to inhibit rotavirus replication in cell culture, toward a long term goal of discovering natural compounds that may complement existing vaccines. Epithelial cells were treated with GA or GRA various times pre- or post-infection and virus yields were measured by immunofluorescent focus assay. Levels of viral proteins VP2, VP6, and NSP2 in GRA treated cells were measured by immunoblot to determine if there was an effect of GRA treatment on the accumulation of viral protein. GRA treatment reduced rotavirus yields by 99% when added to infected cultures post-- virus adsorption, whereas virus yields in GA treated cultures were similar to mock treated controls. Time of addition experiments indicated that GRA-mediated replication inhibition likely occurs at a step or steps subsequent to virus entry. The amounts of VP2, VP6 and NSP2 were substantially reduced when GRA was added to cultures up to two hours post-entry. GRA, but not GA, has significant antiviral activity against rotavirus replication in vitro, and studies to determine whether GRA attenuates rotavirus replication in vivo are underway.