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- 三黃瀉心湯及相關成分抑制A型流行性 感冒病毒及呼吸道融合病毒複製及誘導 免疫機轉探討之體外研究(2-1)
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題 名 | 三黃瀉心湯及相關成分抑制A型流行性 感冒病毒及呼吸道融合病毒複製及誘導 免疫機轉探討之體外研究(2-1)=The Mechanism of the Inhibitive Replication and Inducible Immunization of San-Ou-Say-Sin-To and Its Related Ingredients in Influenza A Virus and RSV in Vitro(2-1) |
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作 者 | 林俊清; | 書刊名 | 中醫藥年報 |
卷 期 | 28:7 2010.09[民99.09] |
頁 次 | 頁267-298 |
分類號 | 414.32 |
關鍵詞 | 三黃瀉心湯; 吸附; 穿透; San-Ou-Say-Sin-To; Attachement; Penetration; |
語 文 | 中文(Chinese) |
中文摘要 | 本研究目標乃從生命科學發展迅速的分子生物學理論與傳統中國醫藥方 劑之作用相互配合,深入探索其抗病毒之活性機轉,如此不僅可增加西方醫 學對中國醫藥作用的接受度,並賦與中國醫藥複方藥效的現代化意義。病毒 感染目前為威脅人類健康的頭號敵人,雖然全世界各國都投入大量的資源進 行研究,但目前仍有許多困難仍需克服:其中包括許多臨床藥物不能完全殲 滅病毒傳染。再者,對於一些具高度突變的病毒會發展出高度的抗藥性,造 成治療的失敗。世界衛生組織 (WHO) 在2001年發表的報告中指出,RSV會引 起18,000-75,000住院病人的感染,而且每年會造成1900人的死亡,根據CDC 對全球的推斷,年齡大的感染人數是嬰幼兒感染人數的四倍,每年大約2.4到 4百萬人會被 感染,因此RSV被指定為主要治療及研究的目標。A型流感病毒 與呼吸道融合病毒對目前臨床用藥敏感性都不高,治療效果通常也不佳。所以,尋求一個能有效且安全的治療藥物乃當務之要。 藥物對細胞增生的抑制活性,抗病毒活性試驗,抑制病毒繁殖的劑量與時間效應,抑制病毒融合作用,抑制病毒吸附試驗與抑制病毒穿透試驗以XTT方式測得。除此之外,抑制病毒融合蛋白分析以免疫分析法測得。 研究發現傳統方劑三黃瀉心湯與其相關成分對常感染人類呼吸道感染的病毒如呼吸道融合病毒與A型流感病毒在不影響宿主細胞生長情況下具有良 好抑制病毒感染效果。但根據研究數據發現三黃瀉心湯與其相關成分抑制呼 吸道融合病毒比A型流感病毒效果較佳。而在三黃瀉心湯相關組成裡發現, 以大黃醇抽物對病毒的作用最好,且大黃相關純品裡又以大黃素對病毒的抑 制最好。除此之外,在藥物抑制病毒時間點發現,大黃醇抽在病毒感染前加 入病毒、感染同時加入病毒或是病毒感染後加入皆發現,此藥物有良好的抑制活性。且隨著劑量增加其抑制作用更為顯著。相同的實驗裡發現大黃素則 是對病毒感染前與同時感染加入藥物的抑制情況最好。從此初步結果推測, 大黃醇抽物之所以對病毒感染前與同時感染有良好作用或許與大黃素有關, 所以接下來皆以大黃醇抽物與大黃素做詳細機轉的探討。在病毒吸附、穿透與融合試驗中結果發現,大黃醇抽物與大黃素隨著劑量增加對病毒有良好的抑制作用。且利用免疫螢光分析大黃醇抽物與大黃素對病毒融合蛋白的影響 發現,此藥物對病毒有明顯抑制作用。 綜合種種的研究發現,三黃瀉心湯與相關成分對呼吸道融合病毒具有良 好抑制作用,由於在病毒感染前後皆有明顯療效,表示此藥物具有預防與治 療作用。建議此後可以進一步發展為臨床藥物。 |
英文摘要 | The incidence of antiviral is increasing worldwide and it is the single most common cause of deaths in both developed and developing countries. World Health Organization (WHO) point out in 2001, RSV will cause 18,000-75,000 impatients' infection, and will cause 1900 people's death every year. According to the data from CDC, the older number of the infected is the quadruple of infant and pre-school children's number of the infected about 2.4 to 4 million people will be infected every year. RSV is appointed as the goal treated and studying mainly.Our laboratory focuses on the integration of traditional Chinese traditional medicines (CTM) with molecular biology to further study the active mechanism their anti-viral activity. This allows a better appreciation of CTM in the modern era and also a better understanding of its underlying potential for therapy, thus increasing its acceptance in Western medicine. Cell proliferation inhibition, antiviral assay, viral replication, viral syncytium, viral attachement and viral penetration were assay by XTT. Besides, viral fusion protein was detect by immunofluorence. Our study reports here for the first time the antiviral effect of SOSST in two upper respiratory tract virus, respiratory syncytial virus and influenza A virus. The results indicated that SOSST and associated crude extract or pure compound successfully inhibited RSV and influenza A virus infection in Hep2 (laryngeal carcinoma cells) and MDCK (Monkey kidney epithelial cell) without significantly affecting cell viability and growth. However, the effect of RSV was batter than influenza A virus. Interestingly, our initial results from the time-of-addition studies indicated that RO (M) could inhibit RSV infection whether added at times of pre-infection (-1 to -4 h), infection (0 h), or post-infection (1 to 4 h). However, emodin inhibited RSV infection was pre- batter than both and post-infection. We showed that RO (M) and emodin could inhibit the RSV attachment penetration and syncytium formation to target cells. In addition, subsequent studies revealed that RO (M) and emodin could also impede RSV post-infection stages by inhibiting viral proteins fusion protein expression. Together, these results suggested that SOSST possesses antiviral activities that can interfere with RSV infection at multiple levels, including directly averting viral attachment and penetration (preventive effect), and inhibiting viral protein (fusion protein) (therapeutic effect). The antiviral mechanisms observed along with its low cytotoxicity level and high selectivity at effective concentrations, suggest that the use of SOSST as direct or supportive treatment for RSV infection merits to be developed. |
本系統中英文摘要資訊取自各篇刊載內容。