大黃與板藍根及其成份抗流行性感冒之分子機制研究

中國醫藥大學機構典藏 China Medical University Repository, Taiwan > 醫學院 > 醫學檢驗生物技術學系暨碩士班 > 研究計畫 > Item 310903500/25658

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题名:	大黃與板藍根及其成份抗流行性感冒之分子機制研究 Investigating the Molecular Mechanisms of Rhubarb, Isatis indigotica and Their Marker Compounds against Influenza a Virus
作者:	林振文
贡献者:	中國醫藥大學醫學檢驗生物技術學系
关键词:	流行性感冒大流行;大黃;板藍根;血球凝集素;神經氨酸脢;M2 蛋白;NS1 蛋白;pandemic influenza;Rhubarb;Isatis indigotica;hemagglutinin;neuraminidase;M2 protein;NS1 protein
日期:	2010-07
上传时间:	2010-09-05 15:44:44 (UTC+8)
摘要:	從禽流感H5N1 在亞洲、非洲及歐洲爆發後，人類流感一直被認為具有嚴重性的全球大流行威脅。目前許多文獻報導顯示世界各國續分離出對治療流感藥物，如adamantine, zanamivir and oseltamivir 具有抗藥性的人類流感及禽流感病毒株。因此，開發新的抗流感藥物與疫苗已成為緊急性課題。本研究計畫目標為藉大黃、板藍根及其主成分等鑑定出具有抑制流感病毒複製的中草藥成分，進而開發以組合微脂粒遞送及抗流感新型預防與治療策略。為達成此目標，首先我們將藉病毒斑點抑制與定量PCR 試驗篩選大黃、板藍根及其主成分抑制流感病毒複製的能力，活性成分再對血球凝集素(HA1、HA3、及HA5 亞型), 神經氨酸脢(NA1 及NA2 亞型), M2 蛋白與NS1 蛋白活性。其次，我們將藉由結合親和力、免疫螢光、血球凝集抑制、神經氨酸脢抑制、干擾素傳遞訊號等方法分析所篩選出的活性成分抑制流感病毒複製的機轉。然後，我們將藉由動物模式探討結合微脂粒遞送抗流感之預防與治療功效。我們所鑑定的抗流感之大黃及板藍根的活性成分可藉由参與阻斷病毒貼附至受器、抑制病毒顆粒釋放、阻斷proton 通道與加強抗病毒免疫反應，達到預防與治療流感之功效。我們預期結果可以將對當前全球流感大流行威脅，貢獻新型且有潛力的預防與治療流感模式。 Pandemic influenza has been recognized as a significant global threat since the emergence of H5N1 avian influenza in domestic poultry flocks in Asia, Africa and Europe. Currently, resistance to antiviral drugs including adamantine, zanamivir and oseltamivir was identified in influenza virus isolates in several countries. New therapies and vaccines against influenza infection become urgent issues. The goal of this proposal is to identify Rhubarb and Isatis indigotica extracts and their maker compounds targeting the influenza viral replication, as well as to develop a combination therapy of Rhubarb and Isatis indigotica extracts and their maker compounds using the liposome delivery system. To achieve this goal, we will identify antiviral activity of Rhubarb and Isatis indigotica extracts and their maker compounds using quantitative methods including the plaque assay and real time RT-PCR. Subsequently, we will determine antiviral properties with in vitro assay systems, such as binding affinity, immunofluorescence, hemagglutination inhibition, neuraminidase inhibition, and IFN signaling. Moreover, we will investigate protective abilities of antiviral herb compounds in liposome delivery system against the lethal challenge with in vivo systems (the mice model). Such assays would be able to identify the herb compounds that are involved in blocking the receptor attachment, inhibiting the release of the progeny virions, blocking proton channel and enhancing antiviral immune responses. Hence, the results obtained from this study may provide valuable information for the combination therapy of herb compound mixtures in liposomes for prophylaxis and therapy against the influenza infection.
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