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目的通过检测蒲公英不同的萃取物体外抗甲型H1N1流感病毒作用,探讨其在甲型H1N1流感治疗上的应用。方法首先用乙醇萃取而得到蒲公英的乙醇萃取物,然后依次采用乙酸乙酯、正丁醇、石油醚、水萃取蒲公英的乙醇萃取物,得到不同溶剂的萃取物;选用狗肾传代MDCK细胞作为病毒的宿主对病毒进行培养和扩增;通过血凝效价和Real time RT-PCR实验,间接或直接检测蒲公英不同萃取物对病毒的中和作用和增殖抑制作用。结果 4种不同溶剂的蒲公英萃取物作用病毒后,只有石油醚、乙酸乙酯组的血凝效价数值出现了一定程度的下降,其中乙酸乙酯萃取物组的值下降了2~4倍;而石油醚萃取物在96 h时,中和实验组和增殖抑制组的血凝效价值分别下降了256倍和128倍。Real time RT-PCR检测表明,石油醚和乙酸乙酯萃取物明显抑制病毒的扩增量。作用96 h时,无论是中和实验还是增殖抑制实验,石油醚萃取物的作用都是最显著的,病毒扩增率较阳性对照分别减少了92.4%和95.6%。结论蒲公英乙酸乙酯萃取物和石油醚萃取物在体外有明显的抗甲型H1N1流感病毒的作用。
Objective To detect the effect of different extracts of dandelion against influenza A (H1N1) virus in vitro and to explore its application in the treatment of influenza A (H1N1). Methods The ethanol extract of dandelion was first extracted by ethanol, then the ethanol extracts of dandelion were extracted by ethyl acetate, n-butanol, petroleum ether and water to obtain the extracts of different solvents. Dogs kidney was used to passage MDCK cells as virus Of the host culture and amplification of the virus; by hemagglutination titer and Real time RT-PCR experiments, indirect or direct detection of different extracts of dandelion on virus neutralization and inhibition of proliferation. Results After the four different solvents of dandelion extracts act as viruses, only the values of blood coagulation potency of petroleum ether and ethyl acetate groups decreased to a certain extent, and the value of ethyl acetate extract group decreased 2-4 times. However, at 96 h of petroleum ether extract, the values of hemagglutination decreased by 256-fold and 128-fold respectively in the neutralization and proliferation inhibition groups. Real time RT-PCR test showed that petroleum ether and ethyl acetate extract significantly inhibited the amplification of the virus. The effect of petroleum ether extract was the most significant at 96 h, both in neutralization and proliferation inhibition experiments. The virus amplification rate was reduced by 92.4% and 95.6% respectively compared with the positive control. Conclusion Dandelion ethyl acetate extract and petroleum ether extract have obvious anti-H1N1 influenza virus effect in vitro.