白及提取物体外抗流感病毒药效及其机理研究  

Anti-influenza Viruse Activity of Extracts from the Tubes of Bletilla striata in vitro

作　　者：张兵 史亚[1] 周芳美[1] 卢亦愚[2] 程东庆[1] ZHANG Bing;SHI Ya;ZHOU Fang-mei;LU Yi-yu;CHENG Dong-qing(Zhejiang Chinese Medical University,Hangzhou 310053,China;Zhejiang Provincial Center for Disease Control and Prevention,Hangzhou 310009,China)

机构地区：[1]浙江中医药大学,浙江杭州310053 [2]浙江省疾病预防控制中心,浙江杭州310009

出　　处：《中药材》2017年第12期2930-2935,共6页Journal of Chinese Medicinal Materials

基　　金：浙江省科技厅公益类项目(2015C33113)

摘　　要：目的:评价白及提取物体外抗流感病毒药效及研究其机理。方法:以病毒血凝效价及核酸量为指标,评价白及提取物在鸡胚模型的抗流感病毒作用。以流感病毒致细胞病变效应(cytopathic effect,CPE)及血凝抑制试验(HI)为指标,评价白及提取物抑制流感病毒的入侵作用及作用靶点。以CPE及病毒RNA合成量为指标,评价白及提取物对病毒复制及RNA合成的抑制作用。神经氨酸酶抑制(NAI)试验测定白及提取物对病毒NA活性的抑制作用。结果:鸡胚模型中,白及提取物各剂量组血凝滴度以及病毒核酸量与病毒对照组比较差异有统计学意义(P<0.01)。细胞实验中,白及水提物和醇提物均具有较好的抗病毒入侵效果,抑制率最高分别达到(87.04±7.81)%和(60.14±6.88)%;水提物通过干预MDCK细胞HA受体阻止病毒的入侵。白及水提物和白及醇提物均具有较好的抗流感病毒复制效果,与病毒对照组比较,白及提取物可显著降低病毒RNA的合成(P<0.01)。醇提物具有较好的神经氨酸酶抑制活性,NA-IC_(50)为16.0 mg/mL。结论:白及提取物通过抑制流感病毒与HA受体结合、干预病毒RNA合成及抑制神经氨酸酶活性发挥抗流感病毒作用。Objective： To evaluate the anti-influenza virus（ H3N2） activity in vitro of Bletilla striata extracts,and to study the mechanisms. Methods： The viral hemagglutination titer and nucleic acid amount were used as measurement indicators to evaluate anti-influenza virus activity in embryonated chicken eggs from aqueous extracts and alcohol extracts of the Bletilla striata. The cytopathic effect（ CPE） and the hemagglutination inhibition（ HI） assay was used to investigate the inhibitory effects and the targets of viral invasion. CPE and the levels of intracellular influenza RNA was carried out to evaluate the inhibition of viral replication and RNA synthesis. The neuraminidase inhibition（ NAI） assay was employed to evaluate the effects of Bletilla striata extracts on the neuraminidase activity（ NA）.Results： The study showed that the viral hemagglutination titer and nucleic acid amount reduced significantly（ P0. 01） compared with the virus control after treated with the extracts of Bletilla striata in embryonated chicken eggs. Both the aqueous extract and alcohol extract had good effect on virus inactivated directly,the highest inhibition rate reached（ 87. 04±7. 81） % and（ 60. 14± 6. 88） % respectively. Aqueous extracts can block the viral attachment by inhibition of MDCK cells viral HA receptor. The aqueous extracts and ethanol extracts were showed effective antiviral activity,compared with virus control,Bletilla striata extracts can significantly reduce the flu virus RNA synthesis（ P0. 01）. Alcohol extracts had good neuraminidase inhibitory activities,NA-IC（50） was 16. 0 mg/mL. Conclusion： The extracts of Bletilla striata can play an anti-influenza virus effect by inhibiting virus HA receptor,and intervene the viral RNA synthesis and neuraminidase activity.

关 键 词：白及 甲型H3N2亚型流感病毒 血凝效价 细胞病变效应法 

分 类 号：R285.5[医药卫生—中药学]