利用雞胚胎蛋建立中藥抗流感病毒之篩選平臺

Establishment of screening platform of traditional Chinese herbal medicine against influenza virus by embryonated chicken egg

蔡佩樺(Pei-Hua Cai)

指導教授 ： 陳志豪

中山醫學大學/醫學院/醫學應用微生物研究所/碩士(2011年)

摘要

流感病毒造成嚴重的全球性公共衛生問題，並可能導致經濟負擔。由於不斷出現抗藥性的流感病毒株，開發新抗病毒藥物是有必要的。中國傳統醫藥（中藥）在亞洲廣泛用於解熱和抗病毒，並予以治療感冒，發燒，慢性支氣管炎。在本研究中利用胚胎雞蛋模式評估333種中草藥的抗病毒活性，利於之後抗病毒藥物的研發。克流感為此模式中的H1N1感染後的有效藥物治療對照組。A型流感病毒感染並於72小時35.5℃培養之後4℃放置隔夜，收集尿囊液測定血凝集效價。候選中草藥利用WST-1測其細胞毒性。經由雞胚胎模式，克流感EC50為 23.7 mg/kg。並從333中草藥中找出22種具抗流感病毒活性的中草藥。這些中草藥包括薔薇科、山茱萸科、薯蕷科、茜草科、菊科、仙茅科、馬兜鈴科、毛茛科、木賊草科、菊科、天南星科、十字花科、玄參科、動物、龍膽科、唇形科、菫菜科、旋花科、百合科。其細胞毒性範圍從0.22 mg/ml 到19.5 mg/ml。總之雞胚胎是一個有利於評估並預測體內抗流感病毒活性的模式。

關鍵字

流感病毒 ； 傳統中藥 ； 雞胚胎

並列摘要

Influenza is a serious global public health problem that can cause economic burdens. Because of continually emerging drug-resistant influenza virus strains, developing new antiviral drugs is critical. Traditional Chinese medicines (TCMs) are broadly used in Asia as antipyretic and antiviral drugs for the treatment of colds, fever, and chronic bronchitis. This study developed an embryonated chicken egg model for evaluating 333 types of antiviral TCMs that act against influenza virus strains. Oseltamivir was used as a positive control against H1N1 in this model. Influenza A virus levels in allantoic fluid were titrated by conducting a hemagglutination test after incubation for 72 h at 35.5 °C and overnight at 4 °C. The cytotoxicity of the candidate TCM was assayed using the WST-1 system and expressed as CC50. Using this model, the EC50 of oseltamivir was 23.7 mg/kg. Twenty-two of 333 tested TCMs exhibited anti-influenza virus activity. The drugs included Rosaceae, Cornaceae, Dioscoreaceae, Rubiaceae, Compositae, Hypoxidaceae, Aristolochiaceae, Ranunculaceae, Equisetaceae, Araceae, Cruciferae, Scrophulariaceae, Gentianaceae, Labiatae, Violaceae, Convolvulaceae, and Liliaceae. The CC50 of the candidate drugs ranged from 0.22 mg/ml to 19.5 mg/ml. In conclusion, this model can be a valuable evaluation of in vivo pretesting of anti-influenza virus activity.

並列關鍵字

influenza virus ； traditional Chinese medicine ； embryonated chicken egg

參考文獻

洪一仁， 廖慧伶， 黃建文，行政院衛生署豐原醫院藥劑科，行政院衛生署豐原醫院胸腔內科。2010年。 對抗H1N1新型流感的新選擇─Peramivir 中南盟臨床專刊 。第3卷。

2009. Swine influenza: frequently asked questions. Weekly Epidemiological Record. World Health Organization. 84:149-160.

Abdul-Careem, M., D. Hunter, M. Lambourne, J. Barta, and S. Sharif. 2007. Ontogeny of cytokine gene expression in the chicken spleen. Poultry science. 86:1351-1355.

Alleva, L.M., C. Cai, and I.A. Clark. 2010. Using complementary and alternative medicines to target the host response during severe influenza. Evidence-Based Complementary and Alternative Medicine. 7:501-510.

Balicer, R.D., M. Huerta, N. Davidovitch, and I. Grotto. 2005. Cost-benefit of stockpiling drugs for influenza pandemic. Emerg Infect Dis. 11:1280-1282.