Title

薑黃素於抑制A型流感病毒感染之作用

Translated Titles

The Effect of Curcumin on Anti-Influenza A virus Infection

DOI

10.6845/NCHU.2008.00923

Authors

陳大元

Key Words

薑黃素 ; 流感 ; 病毒 ; curcumin ; influenza ; virus

PublicationName

中興大學微生物暨公共衛生學研究所學位論文

Volume or Term/Year and Month of Publication

2008年

Academic Degree Category

碩士

Advisor

徐維莉

Content Language

英文

Chinese Abstract

A型流感病毒所引起的流行性感冒持續盛行於世界各地,每年造成龐大的醫療負擔以及經濟損失。由於流感病毒的高度變異性以及高傳染力,如何有效的防治以及治療、避免病毒變異可能引發的新疫情,皆是現在公共衛生研究上的首要工作。A型流感病毒為具有封套的正黏液病毒,其感染細胞起始於表面突起之血球凝集素(Haemagglutinin, HA 蛋白質)與細胞表面受器的結合,經由胞噬作用進入細胞,啟動病毒之複製週期。 研究發現啟動細胞內 nuclear factor-κB (NF-κB)訊息傳遞為維持病毒複製能力所必須。因此,調控 NF-κB路徑的藥物,應能有效的控制流感病毒感染產生的病害。 Curcumin(薑黃素)為 Curcuma longa萃取物之主要成份。不少文獻指出,curcumin具有捕捉自由基、抗發炎、抗病原微生物等能力,以及經由調控細胞內蛋白,例如抑制 NF-κB的表現路徑,來達到抗癌症的效果。有鑒於此,本實驗利用MDCK細胞的模式探討 curcumin於 A型流感病毒的作用。 結果顯示,細胞前處理 curcumin可抑制流感病毒之複製,而且當病毒先與 curcumin作用後,則其感染力盡失; 此現象顯示 curcumin除了藉由調控細胞訊息傳遞路徑之外,也可能經由與A型流感病毒直接作用,而阻擋病毒顆粒貼附或進入細胞,達到抑制A型流感病毒的效果。 以血球凝集抑制試驗(Haemagglutinin inhibition test, HI test)進一步確認 curcumin的抗流感病毒活性可能藉由阻止HA蛋白質的功能。 此外並以 Raman光譜儀初步證實 curcumin與流感病毒表面具有交互作用。 最後以 HI test以及抑制病毒斑形成的能力比較 curcumin的衍生物(curcuminoids) 以及結構類似物(迷迭香酸、薑辣素、辣椒素及異黃酮素)的抗流感作用,結果顯示三種 curcuminoids皆具有抑制HA以及流感病毒的能力,但其餘結構類似物則無。因此推測兩端芳香環數目,分子立體構型,或者其中間碳鏈上的雙鍵數目,可能影響 curcumin與流感病毒的 HA 蛋白質作用,進而影響病毒與細胞受器接合,達到抑制感染的效果。

English Abstract

High pathogenic avian influenza (HPAI) is one of the most important issues in public health nowadays. However, the commonly used prevention or therapeutic strategies have emerged serious problems such as the shortage of vaccine production and the development of drug resistance. Hence, it is of importance to develop new antiviral therapeutic strategies. In present study, the anti-Flu activity of curcumin was evaluated regarding its inhibitory effect on NF-κB pathway which is a prerequisite for efficient influenza replication. Our results demonstrated that pre-treatment of curcumin reduced the yield of IAV progenies to 20% of that of untreated control. Strikingly, results of plaque reduction assay indicated that pre-incubation of virus with curcumin at concentration of 30uM completely abolished the plaque formation in a dose dependent manner. These findings suggest that the anti-flu effect of curcumin seems not only via the effect on cellular level, but also through the direct interaction with the viral protein(s) involved in early events of IAV infection. As the initial stage of IAV infection begins at the binding of viral HA protein to the cellular receptor, the accessibility of HA was further evaluated by means of HA-inhibition (HI) test. Results of the HI test provided evidence that the anti-flu activity of curcumin may via the interruption of HA protein function. By Raman microscopy, a peak formation specific to curcumin pre-incubated influenza virus was revealed indicating a possible interaction between virus particle and curcumin. In addition, to map out the crucial structure involved in anti-flu activity, three curcuminoids that differ by methoxyl groups and four nature structure analogues were analyzed by the properties of HA inhibition and plaque reduction. Results indicated that the unique conformation (cis or trans) and central carbon backbone but not methoxyl groups are important for anti-flu effect of curcumin. Unlike Amantadine, blind passages of influenza viruses in the presence of curcumin did not develop drug resistance. Taken together, in MDCK cells, curcumin harbors strong anti-flu activities and the possibly mechanism is via the interruption of the function of viral HA protein, which lead to the failure of receptor-binding ability of influenza viruses.

Topic Category 醫藥衛生 > 基礎醫學
醫藥衛生 > 預防保健與衛生學
獸醫學院 > 微生物暨公共衛生學研究所
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