Application of anemoside B4 serving as EV71 virus inhibitor in preparing hand-foot-and-mouth disease resisting drugs
A technique of Pulsatilla saponin and hand, foot and mouth disease, which is applied in the field of medicine and achieves the effects of safety, reliability and no toxic and side effects.
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[0016] Example 1: Determination of anti-EV71 activity of Pulsatilla saponin B4 in vitro.
[0017] (1) Virus infection
[0018] Infection mode: RD cells (human rhabdomyoma cells), EV71 can infect and replicate in RD cells, causing apoptosis and necrosis of RD cells, EV71 mainly infects human muscle and nerve tissues, and infecting RD cells with EV71 in vitro can simulate hand, foot and mouth disease model.
[0019] Infection method: EV71 infects RD cells to cause RD cell apoptosis and necrosis. RD cells were planted in 6-well plates. After the cells adhered to the wall, the concentration reached 80%. EV71 infected cells, and normal cultured RD cells were used as blank control. Samples were taken at different times after infection, the cytopathic effect (CPE) of the virus was observed with an inverted fluorescence microscope, and the apoptosis and necrosis of the cells were measured by flow cytometry. The results of flow cytometry are shown in the attached instructions figur...
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[0027] Example 2: Determination of anti-EV71 activity of Pulsatilla saponin B4 in vivo
[0028] (1) Experimental animals: 2-day-old ICR mice with an average body weight of 1.6-1.8 grams.
[0029] (2) Infection method: Take 2-day-old suckling mice and inject EV71 virus intraperitoneally after 5 hours of fasting.
[0030] (3) Experimental grouping: divided into 9 groups: negative group (uninfected virus group), model group (infected with virus, intraperitoneal injection of equal volume of normal saline every day), positive control group (after infected with virus, daily ribavirin Intraperitoneal injection treatment, 100mg / kg / day), experimental group I (after 24 hours of virus infection, intraperitoneal injection of Pulsatilla saponin B41 times a day, the doses were 50, 100, 200mg / kg / day), experimental group II (after 72 hours of virus infection, Daily intraperitoneal injection of Pulsatilla saponin B41 times, the doses were 50,100,200mg / kg / day). There were 12 experimental anim...
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