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Application of Danjie emodin in preparation of medicine for preventing and treating African swine fever virus

A technology of African swine fever virus and emodin, which is applied in the field of biomedicine, can solve the problem of no virus application and achieve good application prospects

Active Publication Date: 2021-12-17
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no application of it in the prevention and treatment of viruses

Method used

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  • Application of Danjie emodin in preparation of medicine for preventing and treating African swine fever virus
  • Application of Danjie emodin in preparation of medicine for preventing and treating African swine fever virus
  • Application of Danjie emodin in preparation of medicine for preventing and treating African swine fever virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Cytotoxicity of Emodin to PAM Cells

[0034] 1. Experimental method

[0035] 1. Preparation of emodin

[0036] Dissolve emodin in DMSO to make 20mM, dilute it with cell culture medium RPMI-1640 when used, wrap it in tinfoil after aliquoting, and store it at -20°C for later use.

[0037] 2. Effect of Danye-emodin on PAM cells

[0038] The PAM cells were seeded in 96-well plates, and after the cells adhered to the wall, the culture medium in the plate was discarded, and RPMI Medium 1640 medium with 10% FBS (containing penicillin 100 U / mL, streptomycin 50 μg / mL, amphotericin 0.25 μg / mL) dilute emodin to 0, 20, 40, 80, 160 and 320 μM, 100 μL / well, set blank cell control group, do 6 duplicate wells for each concentration, at 37 ° C, 5% CO 2 After culturing in the incubator for 48 hours, discard the supernatant, wash twice with PBS, add new culture solution, and then add CCK-8 reagent, 10 μL / well, continue to incubate in the dark for 1 hour, use a microplate read...

Embodiment 2

[0041] Effect of different concentrations of emodin of embodiment 2 on ASFV p30 protein expression in PAM cells

[0042] The p30 protein is a structural protein of African swine fever, which is expressed in the early stage and has a high content, so the p30 protein is selected as a marker for detecting African swine fever virus.

[0043] 1. Experimental method

[0044] Inoculate PAM cells in a 96-well plate, wait for the cells to adhere to the wall, discard the culture medium, inoculate ASFV (MOI=0.1), 100 μL / well, culture at 37°C for 2 hours, discard the virus supernatant, and wash twice with PBS , add 5 μM, 10 μM, 20 μM emodin culture solution respectively, 100 μL / well, at the same time, set the cell group as the negative control and the virus group as the positive control, and set three parallels for each test concentration. Placed at 37°C, 5% CO 2 After culturing in the incubator for 48 hours, discard the supernatant, add 100 μL of 4% paraformaldehyde to each well, fix a...

Embodiment 3

[0047] Effect of different concentrations of emodin of embodiment 3 on ASFV B646L gene expression in PAM cells

[0048]ASFV-B646L is the gene of p72 protein, the main structural protein of African swine fever virus. This gene is highly conserved and must be detected for virus detection.

[0049] 1. Experimental method

[0050] Inoculate PAM cells in a 24-well plate, discard the culture medium after the cells adhere to the wall, add ASFV (MOI=0.1), set a blank cell control group, 500 μL / well, culture at 37°C for 2 hours, discard the virus supernatant solution, washed twice with PBS, added different concentrations of emodin, set virus control group, 500 μL / well, placed at 37 ° C, 5% CO 2 Cultivate in the incubator for 48h. After 48 hours, the cell samples were repeatedly frozen and thawed three times at -80°C and 4°C, and the cell supernatant was collected.

[0051] Use the AxyPrep Body fluid Viral DNA / RNA Miniprep Kit kit (Axygen Company) to extract the genome of ASFV from t...

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Abstract

The invention discloses an application of Danjie emodin in preparation of a medicine for preventing and treating African swine fever virus. The Danjie emodin disclosed by the invention has a remarkable effect of resisting the African swine fever virus. The maximum non-toxic dose of the Danjie emodin is 40 mu M, and the Danjie emodin can effectively inhibit infection and proliferation of the African swine fever viruses, can be used as the medicine for preventing and treating African swine fever, and has a very good application prospect in the aspect of preventing and treating the African swine fever.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of emodin in the preparation of medicines for preventing and treating African swine fever virus. Background technique [0002] African swine fever (ASF) is a severe infectious disease with high infectivity, high morbidity and high fatality rate caused by African swine fever virus (ASFV). Its clinical features include fever, skin Symptoms include reddening, necrotizing dermatitis, and severe bleeding in multiple organs. African swine fever virus is the only member of the African swine fever virus family and the African swine fever virus genus. The size of the virus particle is 200-260nm, it is icosahedral, and it is wrapped by a capsule. The genome length is 170-190kb, and it can encode 150-200 kind of protein. The virulence of the virus varies greatly, and it is clinically manifested as acute, subacute, chronic and recessive infection. The disease is curren...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/085A61K36/70A61P31/20
CPCA61K31/085A61K36/70A61P31/20Y02A50/30
Inventor 王衡郭彦辰张桂红龚浪陈洋孙彦阔郑泽中罗依琢
Owner SOUTH CHINA AGRI UNIV
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