Application of honokiol in treating herpes virus infection
A herpes virus infection, herpes virus technology, applied in the direction of antiviral agents, active ingredients of hydroxyl compounds, etc., can solve the problem of no effective treatment methods
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[0087] Preparation and storage of virus: Infect Vero cells with HSV-1. After a few days, collect the cell culture fluid according to the plaque formation in the cells, which is the virus stock solution, centrifuge at 4000rpm for 30 minutes, collect the supernatant, and distribute it in 1.5mL EP tubes medium, stored at -80°C.
[0088] Recovery and infection of the virus: Take out the HSV-1-GFP virus from -80°C, dissolve it at room temperature, and calculate the required virus volume according to the multiplicity of infection of MOI=0.0005, MOI=0.005, MOI=0.001 and MOI=0.01 , remove the virus and mix with the culture medium.
[0089] Aspirate and discard the original culture medium, add 300 μL virus-containing medium to each well for virus infection, gently shake the culture plate back and forth every 15 minutes, so that the virus-containing medium covers the cells, and infect for 1 hour, during which a total of 4 shakes Second-rate;
[0090] A complete medium containing 10 μg...
Embodiment 1
[0099] Toxicity detection of honokiol in Vero cells
[0100] Honokiol has been proven to promote cell apoptosis in a variety of tumors, such as melanoma, myeloma, leukemia and lung cancer, in order to study its cytotoxicity in Vero cells. After treatment of Vero cells with different concentrations of honokiol for 24 hours, cell apoptosis was detected by flow cytometry.
[0101] The result is as figure 1 As shown, low concentrations of honokiol (5 μg / mL and 10 μg / mL) had little effect on the viability (survival) of Vero cells, while cells began to appear apoptosis when treated with 15 μg / mL honokiol (the cell viability is less than 90%, about 88%), and the viability gradually decreases with the increase of the concentration, indicating that the high concentration of honokiol has obvious killing effect on Vero cells, so 10 μg is selected in the follow-up experiment / mL concentration to detect the effect of honokiol on HSV-1 infection.
Embodiment 2
[0103] In Vero cells, honokiol can inhibit the infection of different titers of HSV-1
[0104] When Vero cells are infected with herpes simplex virus-1 (HSV-1-GFP) with green fluorescent protein, GFP will be expressed together with the expression of viral proteins, so the expression of GFP can be used to indicate the infection of cells by the virus degree.
[0105] The result is as figure 2 shown. From figure 2 As can be seen in A-2C, when Vero was infected with virus of different titers (MOI=0.01, 0.001 and 0.005), it can be seen after 24 hours that compared with the control group, 10 μg / mL honokiol Can significantly inhibit the infection of HSV-1, and there is a statistically significant difference. Generally, the infection process of HSV-1 is divided into primary infection, latent infection and recurrent infection. The above results can confirm that honokiol can significantly inhibit the primary infection of HSV-1.
[0106] When using a lower titer virus (MOI=0.0005)...
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