液态发酵虎乳灵芝菌丝体抗A型流感(H1N1)病毒之效用评估

Evaluation of Effectiveness of Liquid Fermented Lignosus rhinocerus Mycelium against H1N1 Influenza Virus Infections

摘要: 本研究以MDCK犬肾上皮感染H1N1流感病毒模式，评估液态发酵虎乳灵芝(Lignosus rhinocerus)菌丝体萃取物之抗A型流感病毒(H1N1)能力，其中使用MTS法测定病毒或萃取物对宿主细胞之毒杀性，再依药物与病毒作用时间点之不同，分别设定为萃取物预处理、共处理以及后处理三组。结果显示酒萃物之保护效果在共处理组明显优于水萃物，因此将同一批次之虎乳灵芝菌丝体酒萃物进行层析纯化，共获得四种分层，分别为水层(Lr H2O)、正丁醇层(LrBtOH)、乙酸乙酯层(Lr EA)以及正己烷层(Lr Hex)。其中，以Lr H2O、Lr Hex两组在共处理试验(分别为p < 0.01, p < 0.001)以及后处理试验(分别为p < 0.01, p < 0.01)中皆可见显著提升细胞存活率之效果。因此两种成分的极性差异悬殊，故推估液态发酵虎乳灵芝菌丝体的酒萃物中可能具有两种抗H1N1流感病毒之功效成分。

Abstract: The purpose of this study was to evaluate the in vitro anti-influenza A virus (H1N1) potential of ex-tracts from liquid fermented Lignosus rhinocerus mycelia and identify the active compounds that may be responsible. Cytotoxicities of Lignosus rhinocerus mycelia extracts were first determined using MTS assay, by means of time-of-addition experiments: pre-treatment, simultaneous treatment, and post-treatment. As the ethanol extract exhibited a higher survival rate than the aqueous extract, it was then further subjected to differential bioguided fractionation. A total of four fractions, namely aqueous (Lr H2O), butanol (LrBtOH), ethyl acetate (Lr EA) and hexane (Lr Hex) were obtained from the ethanol extract of Lignosus rhinocerus mycelia using partitioning process. Among these fractions, Lr H2O and Lr Hex significantly enhanced the cell survival rate in the simultaneous treatment and post-treatment tests. Conclusively, these findings suggest that ethanolic extract from liquid fermented Lignosus rhinocerus mycelia may contain two anti-H1N1 influenza virus compounds through bioguided fractionation, as the polarities of these two components are very different.