阅读说明：本技术 Egcg在制备预防和/或治疗prv感染制剂中的应用、预防和/或治疗prv感染制剂 (Application of EGCG in preparation of preparation for preventing and/or treating PRV infection and preparation for preventing and/or treating PRV infection ) 是由 郇长超 徐伟音 郭停停 高崧 于 2019-10-25 设计创作，主要内容包括：本发明提供了表没食子儿茶酚没食子酸酯在制备预防和/或治疗猪伪狂犬病毒感染制剂中的应用和预防和/或治疗猪伪狂犬病毒的制剂,属于猪伪狂犬病毒感染治疗技术领域,所述药物中表没食子儿茶酚没食子酸酯的浓度为10～100μmol/L。本发明中,所述表没食子儿茶酚没食子酸酯抗猪伪狂犬病毒感染效果非常显著：所述表没食子儿茶酚没食子酸酯可以减少猪伪狂犬病毒引起的细胞病变,抑制猪伪狂犬病毒的吸附、入胞和复制,且可直接杀死猪伪狂犬病毒,更为重要的是表没食子儿茶酚没食子酸酯可以预防和/或治疗猪伪狂犬病毒感染的小鼠,为临床治疗猪伪狂犬病提供了科学可靠的理论依据。(The invention provides application of epigallocatechin gallate in preparation of a preparation for preventing and/or treating porcine pseudorabies virus infection and a preparation for preventing and/or treating porcine pseudorabies virus, and belongs to the technical field of treatment of porcine pseudorabies virus infection, wherein the concentration of epigallocatechin gallate in the medicine is 10-100 mu mol/L. In the invention, the epigallocatechin gallate has a very significant effect on resisting the infection of the porcine pseudorabies virus: the epigallocatechin gallate can reduce cytopathic effect caused by the porcine pseudorabies virus, inhibit the adsorption, cell entry and replication of the porcine pseudorabies virus, and can directly kill the porcine pseudorabies virus, and more importantly, the epigallocatechin gallate can prevent and/or treat mice infected by the porcine pseudorabies virus, thereby providing a scientific and reliable theoretical basis for clinically treating the porcine pseudorabies.)

1. Application of epigallocatechin gallate in preparing preparation for preventing and/or treating porcine pseudorabies virus infection is provided.

2. The use according to claim 1, wherein the epigallocatechin gallate is used for preventing and/or treating porcine pseudorabies by inhibiting porcine pseudorabies virus adsorption, entry and replication.

3. The use according to claim 1, wherein the epigallocatechin gallate prevents and/or treats porcine pseudorabies by directly killing porcine pseudorabies virus.

4, preparations for preventing and/or treating porcine pseudorabies virus infection, which is characterized in that the concentration of epigallocatechin gallate in the preparations is 10-100 mu mol/L.

5. The preparation according to claim 5, wherein the concentration of epigallocatechin gallate in the medicament is 20-80 μmol/L.

6. The preparation according to claim 5, wherein the concentration of epigallocatechin gallate in the medicament is 40-60 μmol/L.

7. The formulation of claim 5, wherein the solvent for the drug is PBS buffer or DMSO.

8. The preparation of claim 8, wherein the concentration of the PBS buffer is 0.05-0.15 mol/L.

9. The preparation of claim 9, wherein the concentration of the PBS buffer is 0.08-0.12 mol/L.

10. The formulation according to any , wherein the formulation is obtained by dissolving epigallocatechin gallate in a solvent.

Technical Field

The invention belongs to the technical field of treatment of porcine pseudorabies virus infection, and particularly relates to application of epigallocatechin gallate in preparation of a preparation for preventing and/or treating porcine pseudorabies virus infection and a preparation for preventing and/or treating porcine pseudorabies virus.

Background

The porcine pseudorabies virus is a linear double-stranded DNA molecule, belongs to herpesviridae and porcine herpesviridae, and has stronger resistance to the external environment because the outside of virus particles are wrapped by a capsule membrane. Pseudorabies virus is pantropic, can proliferate in a variety of tissue culture cells, is most sensitive to rabbit and pig kidney cells (including primary I cells and continuous cell lines), and causes significant cytopathic effects. Pigs are the storage host of pseudorabies virus, and sick pigs, virus-carrying pigs and virus-carrying mice are important infection sources of the pseudorabies virus. In the swine herd, the virus is transmitted mainly through nasal secretions, and in addition, milk and semen are possible modes of transmission. After pseudorabies happens to the pigs, the clinical symptoms of the pigs are different due to the age of the pigs in days, and the death rate of piglets within 15 days can reach 100 percent; infertility, miscarriage, etc. in pregnant sows; boars exhibit testicular swelling, atrophy, loss of reproductive function, etc. Causing huge economic loss to the pig industry.

Pseudorabies (PR) is a disease caused by a pig herpes virus type 1 (Suid herpesvirus 1, SuHV-1) or Pseudorabies virus (PRV) which is an acute virus infectious disease characterized by fever, severe itch (except pigs), respiratory system and nervous system diseases and encephalomyelitis, and is an outbreak disease in pigs, can cause abortion of pregnant sows, dead fetuses, male pig sterility, mass death of newborn piglets, difficult breathing, growth retardation and the like of pigs, is an important infectious disease which endangers the global pig industry, is an important infectious disease, and in many areas since 2011, a very serious Pseudorabies disease is outbreak, and Bartha-K6332 is inoculated in the pig farms, a newly-appearing PRV strain is very different from PRV strain, and can not provide a great protection effect for pig infection of swine, especially for pig pandemic pig, and the like, so the Pseudorabies virus strain can not provide a great protection effect for pig breeding and pig infection, and the PRV strain can not provide a great protection effect for pig infection.

At present, no effective treatment measures for the variant PRV exist in China, and the occurrence of the variant PRV can be controlled only by early prevention and strengthening sanitary management. In order to reduce the economic losses in the swine industry, there is an urgent need to develop effective drugs for the prevention and treatment of mutated PRV.

Disclosure of Invention

In view of the above, the present invention aims to provide an application of epigallocatechin gallate in preparation of a preparation for preventing and/or treating porcine pseudorabies virus infection, wherein the preparation has characteristics of low cost, no side effect, and significant treatment effect.

In order to achieve the above purpose, the invention provides the following technical scheme:

the invention provides application of epigallocatechin gallate in preparation of a preparation for preventing and/or treating porcine pseudorabies virus infection.

Preferably, the epigallocatechin gallate is used for preventing and/or treating porcine pseudorabies by inhibiting porcine pseudorabies virus adsorption, cell entry and replication.

Preferably, the epigallocatechin gallate prevents and/or treats porcine pseudorabies by directly killing the porcine pseudorabies virus.

The invention provides preparations for preventing and/or treating porcine pseudorabies virus infection, wherein the concentration of epigallocatechin gallate in the preparations is 10-100 mu mol/L.

Preferably, the concentration of epigallocatechin gallate in the preparation is 20-80 mu mol/L.

Preferably, the concentration of epigallocatechin gallate in the preparation is 40-60 mu mol/L.

Preferably, the solvent of the formulation is PBS buffer or DMSO.

Preferably, the concentration of the PBS buffer solution is 0.05-0.15 mol/L.

Preferably, the concentration of the PBS buffer solution is 0.08-0.12 mol/L.

Preferably, the formulation is obtained by dissolving the epigallocatechin gallate in a solvent.

The epigallocatechin gallate has the beneficial effects that the epigallocatechin gallate has a very remarkable effect of resisting the infection of the porcine pseudorabies virus, can reduce cytopathy caused by the porcine pseudorabies virus, inhibits the adsorption, cell-entering and replication of the porcine pseudorabies virus, can directly kill the porcine pseudorabies virus, provides a scientific and reliable theoretical basis for clinically treating the porcine pseudorabies, is a bioactive polyphenol with the most abundant content found in a solid green tea extract, has the advantages of drug source flood, easily obtained raw materials, low cost and the like, is derived from traditional Chinese medicines, has small environmental pollution after being metabolized by an animal body, has no toxic or side effect on the animal, has various biological meanings clinically, and has pharmacological effects of resisting oxidation, arteriosclerosis, thrombosis, angiogenesis, tumor and the like.

Drawings

FIG. 1 is a microscopic observation showing the effect of EGCG on cytopathic effect caused by porcine pseudorabies virus infecting PK-15B6 cells;

FIG. 2 Effect of EGCG on porcine pseudorabies virus infection on PK-15B6 cells; wherein, A is Westernblot for measuring the influence of EGCG on porcine pseudorabies virus infection on PK-15B6 cells; b is TCID50 to determine the effect of EGCG on the infection of porcine pseudorabies virus on PK-15B6 cells; c is the influence of EGCG on the porcine pseudorabies virus infection on PK-15B6 cells measured by fluorescent quantitative PCR;

FIG. 3 is a graph showing the effect of EGCG on porcine pseudorabies virus adsorption and entry cells on PK-15B6 cells, wherein A is Westernblot to determine the effect of EGCG on porcine pseudorabies virus adsorption and entry cells on PK-15B6 cells; b is TCID50 to determine the effect of EGCG on the adsorption and entry of porcine pseudorabies virus on PK-15B6 cells; c, determining the influence of EGCG on porcine pseudorabies virus adsorption and entry cells on PK-15B6 cells by fluorescent quantitative PCR (polymerase chain reaction);

FIG. 4 shows the effect of EGCG on porcine pseudorabies virus adsorption on PK-15B6 cells; wherein A is Western blot for determining the influence of EGCG on porcine pseudorabies virus adsorption on PK-15B6 cells; b is TCID50 to determine the effect of EGCG on adsorption of porcine pseudorabies virus on Dulac cells; c is the influence of EGCG on the porcine pseudorabies virus adsorption on PK-15B6 cells measured by fluorescent quantitative PCR;

FIG. 5 shows the effect of EGCG on porcine pseudorabies virus entry on PK-15B6 cells; wherein A is Weatern blot for determining the influence of EGCG on porcine pseudorabies virus entry on PK-15B6 cells; b is the influence of EGCG on porcine pseudorabies virus encytosis on PK-15B6 cells measured by fluorescent quantitative PCR; c is TCID50 to determine the effect of EGCG on porcine pseudorabies virus entry on PK-15B6 cells;

FIG. 6 shows a Western blot to determine the effect of EGCG on porcine pseudorabies virus replication on PK-15B6 cells;

FIG. 7 is the direct killing effect of EGCG on porcine pseudorabies virus; wherein A is Western blot for measuring the direct killing effect of EGCG on the porcine pseudorabies virus, and B is TCID50 for measuring the direct killing effect of EGCG on the porcine pseudorabies virus;

fig. 8 is a graph demonstrating the protection rate of EGCG against challenge mice by their survival rate.

Detailed Description

The invention provides application of epigallocatechin gallate in preparation of a preparation for preventing and/or treating porcine pseudorabies virus infection. In the invention, the epigallocatechin gallate has a very significant effect on resisting the infection of the porcine pseudorabies virus: the epigallocatechin gallate can reduce cytopathic effect caused by porcine pseudorabies virus; inhibiting the adsorption, cell entering and replication of the porcine pseudorabies virus; and can directly kill the porcine pseudorabies virus; through in vivo test in mice, epigallocatechin gallate can prevent and/or treat mice infected by porcine pseudorabies virus. The source of the epigallocatechin gallate is not particularly required, and the epigallocatechin gallate can be prepared by conventional commercial products in the field or by self.

The invention also provides preparations for preventing and/or treating porcine pseudorabies virus infection, wherein the concentration of epigallocatechin gallate in the preparation is 10-100 mmol/L, preferably 20-80 mmol/L, more preferably 40-60 mmol/L, and most preferably 50 mmol/L.

In the present invention, the drug is preferably obtained by dissolving the epigallocatechin gallate in a solvent. The method and time for dissolving are not particularly limited, and sufficient dissolving can be achieved.

The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.