四川农业大学学报 ›› 2019, Vol. 37 ›› Issue (04): 490-496.doi: 10.16036/j.issn.1000-2650.2019.04.009

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金银花“华金6号”新品种体外抗病毒活性研究

李哲1,2, 玄静1,2, 赵振华1, 马婷1, 张永清1,2,*   

  1. 1.山东中医药大学,济南 250355;
    2.山东省高校中药资源学重点实验室,济南 250355
  • 收稿日期:2018-11-06 出版日期:2019-08-28 发布日期:2021-01-31
  • 作者简介:李哲,硕士研究生。*责任作者:张永清,博士,教授,博士生导师,从事中药资源及质量控制研究,E-mai:zyq622003
  • 基金资助:
    国家公益性行业(农业)科研专项“我国水生、耐盐中药资源的合理利用研究”(201407002); 国家科技重大专项“中药与天然药物活性物质高效分离纯化单元技术平台”(2010ZX09401-302-5-12); 经方常用道地药材培育及质量控制精准化关键技术研究(2016CYJS08A01-2)

Study on the Antivirus Effect of Lonicerae japonicae Flos New Variety‘Huajin 6’ in Vitro

LI Zhe1,2, XUAN Jing1,2, ZHAO Zhenhua1, MA Ting1, ZHANG Yongqing1,2,*   

  1. 1. Shandong University of Traditional Chinese Medicine,Jinan 250355,Shandong,China;
    2. Key Laboratory of Chinese Medicine Resources in Colleges and Universities of Shandong Province,Jinan 250355,Shandong,China
  • Received:2018-11-06 Online:2019-08-28 Published:2021-01-31

摘要: 【目的】探讨金银花“华金6号”新品种体外抗H9-AIV活性。【方法】水煎提取,所得药液终浓度为1 g/mL,HPLC法测定绿原酸含量,鸡胚尿囊腔接种及血凝(HA)试验检测体外抗H9-AIV活性。根据金银花水提液的最大不凝血浓度,将“华金6号”新品种、平邑“大毛花”水提液稀释成31.25、15.625、7.812 5 mg/mL 3个浓度,每个浓度设4组平行对照,进行H9-AIV的体外直接抑制试验;将“华金6号”新品种、平邑“大毛花”水提液从对鸡胚的最大无毒浓度(TDO)起以2倍比稀释成125、62.5、31.25 mg/mL 3个浓度,每个浓度接种6枚鸡胚,0.2 mL/胚,分别进行中和试验、预防试验、治疗试验,并设病毒(100EID50)、利巴韦林(50 mg/mL)及PBS缓冲液对照组。【结果】“华金6号”新品种绿原酸含量为0.93%,而传统“大毛花”品种为0.91%,二者无显著差异;“华金6号”新品种与“大毛花”品种3个浓度组体外直接抑制H9-AIV的活性均较强,与对照组差异显著(P<0.05),“华金6号”新品种略优于“大毛花”金银花,且二者抗H9-AIV活性在24 h时达到最高;将金银花提取液与病毒体外先中和再接种时,两种金银花均表现为高浓度(125 mg/mL)抑制活性高、低浓度(62.5、31.25 mg/mL)抑制活性低;“华金6号”新品种与“大毛花”金银花预防与治疗H9-AIV效果无显著差异(P>0.05)。【结论】“华金6号”新品种与“大毛花”金银花均有较强的体外直接抑制H9-AIV活性,“华金6号”抗病毒活性略高于“大毛花”金银花。

关键词: 金银花, 华金6号, 抗病毒

Abstract: 【Objective】 To study the anti-H9AIV effect of Lonicerae japonicae Flos New Variety ‘Huajin 6’ in vitro. 【Method】 Lonicerae japonicae Flos were extracted by water decoction and the final concentra- tion of the obtained solution was 1 g crude drug /ml, the content of chlorogenic acid was assayed by HPLC; SPF embryo inoculation method and hemagglutination (HA) test were adopted in this research to detect antiviral activity in vitro.According to the maximum non-coagulation concentration of the aqueous extract of honeysuckle, the aqueous extract of the new variety‘Hua jin 6’and‘Da mao hua’were diluted into 31.25,15.625,7.812 5 mg/mL 3 concentrations. Four parallel control groups were set for each concentration to conduct direct inhibition test of H9-AIV in vitro. From the maximum nontoxic concentration (TDO), the two kinds of honeysuckling water extracts were diluted into 3 concentrations(125, 62.5, 31.25 mg/mL), each concentration was inoculated with 6 chicken embryos, 0.2 mL/embryo,respectively, for neutralization test, prevention test, treatment test, and virus (100EID50), ribavirin (50 mg/mL) and PBS buffer solution control group. 【Result】 The content of chlorogenic acid was 0.925% and 0.914% in the extract sample of Lonicerae japonicae Flos New Variety‘Hua jin 6’and ‘Da mao hua’. In vitro, two kinds of honeysuckle both had a strong effect on anti-H9AIV and the effect of ‘Hua jin 6’ was slightly better, there was significant difference between the honeysuckle group and the viral control group (P<0.05). The anti-H9AIV effect was the best in 24h; When the drug is neutralized with the virus and then inoculated with chicken embryo, the two kinds of honeysuckle showed high inhibitory activity at high concentration (125 mg/mL) and low inhibitory activity at low concentration (62.5, 31.25 mg/mL) ; In the prevention and treatment of H9-AIV, there was no significant difference between the new variety of ‘Hua jin 6’ and ‘Da mao hua’ (P>0.05) . 【Conclusion】 In vitro antivirus effect,two kinds of honeysuckle both had strong effect on the direct inhibition of H9-AIV,and Lonicerae japonicae Flos ‘Hua jin 6’ new variety was slightly better than ‘Da mao hua’.

Key words: Lonicerae japonicae Flos, ‘Hua jin 6’, antiviral activity

中图分类号: 

  • R285.5