A plant source preparation for antiviral infection of livestock and poultry

An anti-virus, livestock and poultry technology, applied in the field of plant-derived preparations for anti-viral infection of livestock and poultry, can solve problems such as unclear anti-viral effect, and achieve the effect of broad-spectrum and high-efficiency anti-viral activity

Inactive Publication Date: 2021-02-09
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported in the early years that HHT can inhibit the infection of hepatitis B virus and mouse hepatitis virus to a certain extent, but the mechanism of action and the antiviral effect in vivo are still unclear.

Method used

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  • A plant source preparation for antiviral infection of livestock and poultry
  • A plant source preparation for antiviral infection of livestock and poultry
  • A plant source preparation for antiviral infection of livestock and poultry

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0031] 1. Preparation of cells

[0032] DMEM medium containing 10% fetal bovine serum and 100U penicillin and streptomycin was used for HeLa, Vero, PK15 and Marc145 cell passage, and DMEM medium containing 2% fetal bovine serum and 100U penicillin streptomycin was used as maintenance medium.

[0033] NDV infected HeLa cells, VSV, HSV-1, PEDV, PRV infected Vero cells, TGEV, FMDV infected PK15 cells, PRRSV infected Marc145 cells.

[0034] 2. Virus proliferation

[0035] Proliferation of HSV-1, NDV, VSV, and PRV cytotoxicity: Inoculate the virus stock solution according to the amount of 1 / 10 of the cell flask culture medium, and wait for the cytopathic changes (such as NDV to form large syncytia) to reach 75% (usually 48 hours) to use repeatedly Harvest the virus by freezing and thawing, that is, store the cell bottle at -80°C and store it at room temperature until it partially thaws. The virus was released from the cells and frozen at -80°C.

[0036] Proliferation of NDV chic...

Embodiment 1

[0043] Embodiment 1 The influence of HHT on the virulence of VSV virus

[0044] Infect HeLa cells with 0.5MOI of VSV, add different concentrations of HHT and Ribavirin (Ribavirin) to the cell culture medium after 1.5h, freeze and thaw three times after infecting 24h, measure virus virulence, the results show that compared with Ribavirin, the effective antiviral concentration of HHT is 1000 times less ( figure 2 A).

[0045] HeLa cells were infected with 1 MOI of VSV, 1.5 hours later, different concentrations of HHT were added to the cell culture medium, and the cells were collected after 36 hours of virus infection. The expression of viral protein G protein was detected by immunoblotting, and the results showed that HHT significantly inhibited the expression of G protein in a dose-dependent manner. HeLa cells were infected with 0.5 MOI of VSV, and 50nM HHT was added after 1.5h, and the cell samples were collected at 8h, 24h, and 36h to detect the expression of viral protein...

Embodiment 2

[0047] Example 2 Effect of HHT on NDV virus virulence

[0048]Infect HeLa cells with 0.1 MOI of GFP-NDV (green fluorescent protein-labeled NDV), add different concentrations of HHT to the cell culture medium after 1.5 h, and the concentration of HHT from low to high is 5, 10, 25, 50 , 100 nM. After virus infection 24h, observe in fluorescent microscope Olympus IX73, image with CellSens software, the result shows that, HHT significantly inhibits GFP-NDV infection and is dose-dependent ( image 3 A).

[0049] HeLa cells were infected with 0.1 MOI of GFP-NDV. After 1.5 hours, different concentrations of HHT were added to the cell culture medium. After 36 hours of virus infection, the cells were collected, and the expression of viral protein NP was detected by Western blotting. The results showed that HHT significantly inhibited the expression of NDV protein and in a dose-dependent manner ( image 3 B).

[0050] HeLa cells were infected with 0.1 MOI of GFP-NDV. After 1.5 hours...

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Abstract

The invention relates to a plant source preparation for antiviral infection for livestock and poultry, in particular to the application of HHT as a broad-spectrum antiviral preparation. The present invention finds for the first time that homoharringtonine (HHT) has broad-spectrum and high-efficiency antiviral activity, and can efficiently inhibit 9 kinds of viruses (VSV, NDV, PEDV, TGEV, AIV, HSV‑1, PRV, PRRSV, FMDV) ) to infect host cells and animals (chicken, chicken embryos, pigs, etc.), significantly reduce the amount of virus in the animal body, and alleviate the symptoms of infection, providing safe and reliable new biological agents for the breeding industry. The invention provides the basis and possibility for developing broad-spectrum and high-efficiency antiviral new drugs, and provides a new safe and reliable technical means for ensuring the sustainable development of agriculture and animal husbandry and human health. It has far-reaching practical significance for developing traditional medical resources and promoting the cause of traditional Chinese medicine in our country.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a plant source preparation for antiviral infection for livestock and poultry. Background technique [0002] At present, for viral infections, especially when the pathogen is sudden or unknown, there are no broad-spectrum, efficient and safe drug treatments similar to antibiotics. In addition, viral biochemical weapons are increasingly becoming a huge potential threat to all human beings. These facts emphasize the new The necessity and urgency of developing broad-spectrum antiviral agents. Vaccine is a recognized and feasible treatment method that has made positive contributions to human health, but it also faces some thorny problems and shortcomings, such as narrow spectrum - the vaccine is only effective against a specific serotype of virus; So much so that the development of vaccines can only be carried out after the onset of infectious diseases; in addition, the development of vacc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/55A61K36/13A61P31/22A61P31/16A61P31/14A61P31/20
CPCA61K31/55A61K36/13
Inventor 王晓佳薛彦沈志强董慧君
Owner CHINA AGRI UNIV
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