黃芩抑制A型流感病毒神經胺酸酶之研究

黃芩（Scutellaria baicalensis Gergi）通常做為傳統中藥複方成分，處方為治療發熱或炎症急病。黃芩在臨床上以證實有許多功效，包括抗病毒、抗過敏、抗發炎、抗細菌、抗腫瘤之效果。A型流感病毒屬於正黏液病毒，為一種高傳染性單股負性RNA病毒，易引起季節性或全球大流行。本論文研究針對黃芩粗抽物及指標成分對抗A型流感病毒功效之探討。首先建立抑制試管內神經胺酸酶活性實驗方法，並分析黃芩抑制病毒神經胺酸酶活性之抑制百分之五十之濃度（50％ inhibiting concentration, IC50）。其次，利用MTT方法分析藥物濃度百分之五十之的毒殺效果（50％ cytotoxicity concentration, CC50），然後觀察黃芩抑制A型流感病毒感染細胞之細胞病變（CPE,Cytopathic effect）於MDCK細胞之情形，再以即時定量聚合酶連鎖反應（Real-time PCR）分析黃芩抑制A型流感病毒之產量，進一步利用病毒斑試驗來分析黃芩對A型流感病毒之抑制百分之五十之濃度（50％ inhibiting concentration, IC50）。抑制季節性A型流感病毒神經胺酸酶活性實驗結果顯示，黃芩甲醇、以酸乙酯層、氯仿層及baicalein、baicalin、apigenin及chrysin之IC50為分別為473.93 μg/ml、 57.14 μg/ml、103.51 μg/ml、0.423 μM、 2.55 μM、61.72 μM及109.64 μM。黃芩乙酸乙酯層粗抽物濃度為100μg/ml，明顯抑制A型流感病毒之細胞病變現象。在以即時定量聚合酶連鎖反應實驗結果顯示，在細胞以藥物及病毒處理後，會造成流感病毒複製倍數下降，黃芩以酸乙酯層粗抽物濃度100μg/ml ΔCt值大於3.3，達到百分之九十有抑制效濃度（90％ effective concentration, EC90）。結果顯示出黃芩乙酸乙酯層粗抽物可以抑制A型流感病毒神經胺酸脢活性、抑制病毒斑現象以及抑制病毒複製情形。本實驗提供黃芩為一個新的抗A型流感病毒之藥物。

關鍵字

黃芩；流感病毒；神經胺酸酶

並列摘要

Scutellaria baicalensis Georgi is often included as an ingredient in traditional Chinese compound prescriptions for the treatment of fever-related or inflammatory conditions. It has been demonstrated to have various clinical activities, including anti-virus, anti-allergic, anti-inflammatory, anti-microbial, and anti-cancer effects. Influenza A virus (IAV) belongs to the Orthomyxovirus family, being segmented, single stranded, negative sense RNA virus. IAV is the important respiratory pathoge, being responsible for both seasonal influenza outbreaks and periodic world-wide pandemics. The goal of this study is to investigate the antiviral effect of Scutellaria baicalensis Gergi and their marker compounds against Influenza A virus (IAV) in vitro assay. First, neuraminidase inhibition activity assay was used to measure for 50% inhibiting concentration (IC50) of each extract and marker compounds. The MTT assay was used to measure for 50% cytotoxicity concentration (CC50) of each extract and marker compounds. To investigate the efficacy of Scutellaria baicalensis Gergi and their marker compounds as antivirals against Influenza A virus by assessing their ability to reduce Influenza A virus RNA levels in MDCK cells. The plaque assay was carried out detecting IC50 of extracts and compounds of Scutellaria baicalensis Gergi. The IC50 values of methanol extract, ethyl acetate extract, chloroform extract, baicalein, baicalin, apigenin and chrysin were 473.93 μg/ml、 57.14 μg/ml、103.51 μg/ml、0.423 μM、 2.55 μM、61.72 μM and 109.64 μM for in vitro neuraminidase inhibition activity assay. Moreover, Scutellaria baicalensis Gergi and its marker compounds inhibited the neuraminidase activity of IAV. Extracts and compounds of Scutellaria baicalensis Gergi showed the inhibitory ability on the IAV-induced CPE in MDCK cells. Otherwise, real-time PCR demonstrated the inhibition of IAV titer by extracts and the marker compounds of Scutellaria baicalensis Gergi. Therefore we concluded that Scutellaria baicalensis Gergi is effective in inhibiting the replication of influenza virus, and it is more likely to be a potential antiinfluenza viral agent for clinical use.

並列關鍵字

Scutellaria baicalensis Gergi ； neuraminidase ； influenza

參考文獻

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黃芩抑制A型流感病毒神經胺酸酶之研究

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