Application of tripterine in preparation of anti-EHV-8 medicine

Abstract

The invention relates to the technical field of traditional Chinese medicine production, in particular to application of traditional Chinese medicine tripterine in preparation of an anti-EHV-8 medicine. Comprising an application of the tripterine in preparation of the anti-EHV-8 medicine, an application of a plant extract containing the tripterine in preparation of the anti-EHV-8 medicine and an application of a traditional Chinese medicine compound containing the tripterine in preparation of the anti-EHV-8 medicine. The research on inhibition of EHV-8 by tripterine is carried out in in-vitro susceptible cells and a mouse model in-vivo system, and the result shows that tripterine can significantly inhibit replication of EHV-8, can be used as an effective drug component for preventing and treating EHV-8 infection, fully plays the advantage of small toxic and side effects of natural compounds of plant sources, and has a good application prospect in prevention and treatment of EHV-8 infection. Valuable theoretical basis and reference are provided for clinical medication of EHV-8, including antiviral and anti-inflammatory effects, and hosts can be protected from dual damage of viruses and inflammations.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine production, and specifically to an application of tripterygium wilfordii in the preparation of an anti-EHV-8 drug. Background Art

[0002] Previous studies have found that equine animals (horses or donkeys) infected with equine herpesvirus type 8 (EHV-8) can cause rhinopneumonia, which can cause breathing difficulties and death in severe cases. Pregnant horses or donkeys are often latently infected, with sudden miscarriages and neurological symptoms in some donkeys. The virus seriously endangers the health of equine animals. EHV-8 was first isolated from the nasal cavity of donkeys in Australia; subsequently, domestic researchers isolated the EHV-8wh strain from horses for the first time in 2010; in 2020, Israeli scholars isolated the EHV-8 wild strain from an adult male donkey for the first time.

[0003] EHV-8 belongs to the alpha herpes virus. The diameter of the virus particle is about 150nm. The icosahedral nucleocapsid with a diameter of 100nm is located inside the virus particle. The structure of the mature virus particle consists of three parts: the virus core, the capsid and the capsule. The genome in the nucleocapsid is linear and reel-like, and the lipoprotein capsule and its small fibrous protrusions composed of glycoproteins are located outside the nucleocapsid. Equine herpes virus is an enveloped double-stranded DNA virus with a genome length of about 150kb, containing 80 open reading frames and encoding 76 proteins.

[0004] In recent years, with the rapid development of the horse and donkey industries, the number of epidemics in large-scale horse and donkey farms has gradually increased, especially the high abortion rate and frequent respiratory diseases, which seriously trouble the sustainable development of the horse and donkey industries. Our team's preliminary research found that EHV-8 is the main pathogen that causes donkey abortion, respiratory diseases and viral encephalitis. Multiple strains of EHV-8 wild virus were isolated from the lungs, placentas and brain tissues of dead donkeys and donkeys. At present, there is no effective vaccine or drug for this virus, and an economical and efficient antiviral drug is urgently needed for the prevention and control of EHV-8.

[0005] Tripterygium wilfordii is a natural pentacyclic triterpenoid compound extracted and separated from the traditional Chinese medicine Tripterygium wilfordii. Its molecular formula is C29H38O4 and its molecular weight is 450.61 (such as Figure 1 As shown). Modern pharmacological studies have shown that the drug has multiple pharmacological activities such as immunomodulation, anti-inflammatory, anti-tumor, anti-fibrosis, protection of cardiovascular and cerebrovascular and nervous systems, and good in vivo safety. In recent years, it has received extensive attention in anti-inflammatory and anti-tumor research due to its advantages such as low adverse reactions, high efficiency, broad spectrum, multiple targets, and low drug resistance. However, there are currently no reports on the use of tripterygium wilfordii to prevent and treat EHV-8 infection. Summary of the invention

[0006] The purpose of the present invention is to provide an application of tripterygium wilfordii in the preparation of anti-EHV-8 drugs to make up for the problem of lack of EHV-8 vaccines and prevention and control drugs.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] The invention discloses an application of tripterygium wilfordii in preparing an anti-EHV-8 drug, including the application of tripterygium wilfordii in preparing a drug for inhibiting EHV-8 replication.

[0009] Preferably, the invention also includes the use of plant extracts containing tripterygium wilfordii in the preparation of anti-EHV-8 drugs.

[0010] Preferably, the invention also includes the use of a traditional Chinese medicine compound containing tripterygium wilfordii in the preparation of anti-EHV-8 drugs.

[0011] Preferably, the tripterygium wilfordii, the plant extract containing tripterygium wilfordii and the traditional Chinese medicine compound containing tripterygium wilfordii are used for preventing and treating respiratory diseases caused by EHV-8 infection in equine animals.

[0012] Preferably, the tripterygium wilfordii, the plant extract containing tripterygium wilfordii and the traditional Chinese medicine compound containing tripterygium wilfordii are used for preventing and treating abortion caused by EHV-8 infection in equine animals.

[0013] Preferably, the tripterygium wilfordii, the plant extract containing tripterygium wilfordii and the traditional Chinese medicine compound containing tripterygium wilfordii are used to treat viral encephalitis caused by EHV-8 infection.

[0014] Preferably, at the susceptible cell level, the maximum safe mass concentration of tripterygium wilfordii is 1 μmol / L.

[0015] Preferably, the tripterygium wilfordii, the plant extract containing tripterygium wilfordii and the traditional Chinese medicine compound containing tripterygium wilfordii all have inhibitory effects on multiple strains of EHV-8 (SDLC66, SD2020113, and donkey / Shandong / 10 / 2021).

[0016] Preferably, the tripterygium wilfordii, the plant extract containing tripterygium wilfordii and the traditional Chinese medicine compound containing tripterygium wilfordii all have inhibitory effects at multiple stages of EHV-8 replication.

[0017] Preferably, the tripterygium wilfordii, the plant extract containing tripterygium wilfordii and the traditional Chinese medicine compound containing tripterygium wilfordii are used to reduce lung damage in a mouse model caused by EHV-8 infection.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. Inhibit viral replication

[0020] The maximum safe mass concentration of tripterygium wilfordii on susceptible cells (RK-13 ​​and NBL-6) is 1 μmol / L, at which it can effectively inhibit viral replication.

[0021] In vitro, the study of EHV-8 susceptible cells RK-13 ​​and NBL-6 found that tripterygium wilfordii had a significant inhibitory effect on the replication of EHV-8SDLC66. qPCR and Western blot analysis showed that tripterygium wilfordii could significantly inhibit EHV-8gD gene transcription and gD protein expression.

[0022] In vitro, using the IFA method, it was confirmed that triptolide has a significant inhibitory effect on the replication of multiple strains of EHV-8 (such as SDLC66, SD2020113, and donkey / Shandong / 10 / 2021, etc.).

[0023] With the help of qPCR and Western blot methods, it was determined that tripterygium wilfordii can exert an antiviral effect in multiple stages of EHV-8 replication (such as pretreatment group, co-treatment group, post-infection incubation group, full-stage treatment group, etc.), but it cannot directly inactivate EHV-8.

[0024] 2. Anti-EHV-8 effect in vivo

[0025] In the mouse model, tripterygium wilfordii can effectively reduce the replication level of EHV-8 in the lungs of BALB / c mice. 50 The results showed that the replication level of EHV-8 in the lungs of mice treated with tripterygium wilfordii was significantly lower than that in the control group.

[0026] 3. Drug advantages

[0027] Tripterygium wilfordii can be used as an effective drug ingredient for the prevention and treatment of EHV-8 infection, giving full play to the advantages of natural compounds derived from plants with low toxicity and side effects.

[0028] Triptolide has anti-inflammatory effects and can reduce the damage of inflammatory responses to host cells. After infection with EHV-8, host cells release a variety of inflammatory factors. Triptolide can protect the host from the dual damage of virus and inflammation by inhibiting the release of inflammatory factors.

[0029] Tripterine has multiple pharmacological activities such as immunomodulation, anti-tumor, anti-fibrosis, and protection of cardiovascular and nervous systems, which provides it with more potential advantages as an anti-EHV-8 drug.

[0030] In summary, the application of tripterygium wilfordii in the preparation of anti-EHV-8 drugs of the present invention systematically carried out tripterygium wilfordii anti-EHV-8 studies in vitro and in vivo in mouse models, revealing that tripterygium wilfordii can significantly inhibit the replication of EHV-8, can be used as an effective ingredient of drugs for preventing and treating EHV-8 infection, and give full play to the advantages of natural compounds of plant origin with small toxic and side effects, providing valuable theoretical basis and reference for the clinical use of EHV-8, including antiviral and anti-inflammatory effects, which can protect the host from dual damage of viruses and inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they provide further detailed explanations but do not constitute a limitation of the present invention.

[0032] Figure 1 This is the molecular formula diagram of tripterygium wilfordii;

[0033] Figure 2 The toxicity test results of tripterygium wilfordii on EHV-8 susceptible cells of the present invention;

[0034] Figure 3 The tripterygium wilfordii extract of the present invention significantly inhibits the infection of EHV-8SDLC66; the gD gene transcription (A) and the expression of gD protein (B) are reduced;

[0035] Figure 4 The results of indirect immunofluorescence analysis (IFA) show that tripterygium wilfordii has an inhibitory effect on different strains of EHV-8.

[0036] Figure 5 Schematic diagram of the experimental setup of different treatment methods of tripterygium wilfordii and EHV-8 infection of the present invention;

[0037] Figure 6 This is a result diagram showing that tripterygium wilfordii of the present invention exerts an inhibitory effect on multiple replication stages of EHV-8 infection;

[0038] Figure 7 This is a result diagram showing that tripterygium wilfordii in the present invention cannot directly inactivate EHV-8;

[0039] Figure 8 HE staining diagram showing that tripterygium wilfordii reduces the damage caused by EHV-8 in the mouse lungs;

[0040] Fig. 9 This is a graph showing the results of the celastrol of the present invention reducing the replication level of EHV-8 in the lungs of mice. DETAILED DESCRIPTION

[0041] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0042] Example 1 Toxicity test of tripterygium wilfordii on EHV-8 susceptible cells

[0043] Rabbit kidney cells RK-13 ​​and horse dermal cells NBL-6 were seeded in 96-well plates (1×10 4 / well), when the confluence reached about 80%, 100 μL of different concentrations (0, 0.1 μM, 0.2 μM, 0.4 μM, 0.8 μM, 1.6 μM, and 3.2 μM) of tripterygium wilfordii dilution was added to each well. At the same time, DMSO negative control and blank control were set up, and 3 replicate wells were set up for each concentration. After incubation in a 37°C, 5% CO2 incubator for 24 hours, 10 μL of CCK-8 solution was added to each well and cultured for 2 hours. With the help of an enzyme reader, the absorbance value at OD450nm was read, and the following formulas were used to calculate the effects of tripterygium wilfordii on the activity of RK-13 ​​and NBL-6 cells, respectively, and the maximum safe mass concentration of tripterygium wilfordii was evaluated (the results are shown in Figure 2 shown).

[0044] Cell viability (%) = [A1-A2] / [A3-A2] × 100%

[0045] A1 test group: absorbance values ​​of wells with cells, CCK-8 solution and tripterygium wilfordii solution

[0046] A2 Blank control: absorbance value of the well with only culture medium and CCK-8 solution but no cells

[0047] A3 Negative control: absorbance value of the well with cells and CCK-8 solution but no tripterygium wilfordii solution

[0048] The results are as follows Figure 2 As shown, different concentrations of tripterygium wilfordii had no significant effect on the viability of RK-13 ​​and NBL-6 cells, and the maximum safe mass concentration was 1 μM.

[0049] Example 2: Tripterine inhibits infection of EHV-8SDLC66 in vitro

[0050] 1. Analysis of EHV-8gD gene expression by qPCR and Western blot

[0051] RK-13 ​​and NBL-6 cells were seeded in 6-well cell plates (2×10 5 / well), when the cells were confluent to 80%, different concentrations of tripterygium wilfordii (0μM, 0.25μM, 0.5μM and 1μM) were added for pretreatment for 2h, and EHV-8SDLC66 (0.1MOI) was inoculated for 1h, the virus solution was discarded, and cell maintenance solution containing tripterygium wilfordii was added. A DMSO-treated group and a normal cell untreated group were set as control groups, respectively. After 24h of virus infection, the cell supernatant was collected, and the total cell DNA was extracted using a kit. The changes in the EHV-8gD virus copy number were determined by qPCR. The results are shown in Figure 3 shown.

[0052] RK-13 ​​and NBL-6 cells were seeded in 6-well plates (2×10 5 / well), when the cells were confluent to 80%, different concentrations of tripterygium wilfordii (0μM, 0.25μM, 0.5μM, and 1μM) were added and cultured for 2h. After inoculation of EHV-8SDLC66 (0.1MOI) for 1h, the virus solution was discarded, and the cell maintenance solution containing tripterygium wilfordii was added. The DMSO-treated group and the normal cell untreated group were set as control groups, and the cells were collected 24h after infection. The changes in the expression level of EHV-8gD protein were analyzed by Western blot. Figure 3 As shown, tripterygium wilfordii significantly inhibited the expression of EHV-8 gD protein.

[0053] 2. Tripterygium wilfordii has an inhibitory effect on multiple EHV-8 strains

[0054] RK-13 ​​cells were inoculated into 12-well cell culture plates. When the cells were confluent to 80%, they were pretreated with different concentrations of tripterygium wilfordii (0.25 μM, 0.5 μM and 1 μM). The DMSO-treated group was used as a negative control. The drug solution was discarded after 2 hours. The cells were infected with EHV-8SDLC66, EHV-8SD2020113 and EHV-8donkey / Shandong / 10 / 2021 for 1 hour, respectively. The supernatant containing drugs and viruses was discarded and replaced with 3% FBSDMEM. After 48 hours of infection, the cells were fixed with 75% ethanol, and the positive serum of mouse anti-EHV-8 was used as the primary antibody. The CY3-labeled sheep anti-mouse fluorescent antibody was used as the secondary antibody. The effect of tripterygium wilfordii on the replication of EHV-8 was evaluated by indirect immunofluorescence experiment (IFA). The results are shown in Figure 4 shown.

[0055] 3. Determine the effect of tripterygium wilfordii on the replication cycle of EHV-8

[0056] To further explore the mechanism of tripterygium wilfordii affecting EHV-8 infection, RK-13 ​​and NBL-6 cells were inoculated in 12-well plates (1×10 5After the cells were confluent to 80%, the control group, the celastrol pretreatment group (Pre), the celastrol and EHV-8 co-treatment group (Co), the celastrol incubated group after EHV-8 infection (Post), and the celastrol full-stage treatment group (All stages-treatment) were set up respectively (see Figure 5 ), inoculated with EHV-8SDLC66 (MOI = 0.1), and 24 h later, cells were collected to detect viral copy number by qPCR and to evaluate gD protein expression by Western blot. Figure 6 As shown. This suggests that tripterygium wilfordii exerts antiviral effects at multiple stages of EHV-8 replication.

[0057] 4. Determine whether tripterygium wilfordii has the activity of inactivating EHV-8

[0058] To further explore the mechanism of tripterygium wilfordii on EHV-8 infection, RK-13 ​​and NBL-6 cells were seeded in 12-well plates (1×10 5 / well), after overnight culture. Tripterygium wilfordii (1μM) was incubated with different doses of EHV-8SDLC66 (0.1MOI, 0.5MOI and 1MOI) at 37℃ for 2h. After infecting the cells for 1h, they were replaced with 3% FBS MEM for 24h, and the cell supernatants were collected to detect the virus copy number by qPCR. The results are shown in Figure 7 This suggests that tripterygium wilfordii cannot directly inactivate EHV-8.

[0059] 5. In vivo study on the effect of tripterygium wilfordii on EHV-8 infection

[0060] Twelve SPF BALB / c mice were randomly divided into 4 groups, 3 mice in each group. A negative control group (Mock); DMSO-treated control group + EHV-8; tripterygium wilfordii (2 mg / kg) + EHV-8; tripterygium wilfordii (4 mg / kg) + EHV-8 were set up. The mice in the tripterygium wilfordii drug-treated group were intraperitoneally injected with the corresponding dose of the drug 1 day before infection, and the mice in the EHV-8 infection control group were injected with the same dose of DMSO solution. Except for the negative control group, the mice in each group were intranasally infected with 100 μL (1×10 5 PFU / mouse) EHV-8SDLC66, and were administered once at fixed time points on the first day after infection (i.e., 1dpi) and the third day (i.e., 3dpi), and their weight changes and status were detected every day. The mice were killed by cervical dislocation on the seventh day after virus infection (i.e., 7dpi), and the lungs were collected for formaldehyde fixation and histopathological examination. The results showed that tripterygium wilfordii could significantly reduce the pathological damage caused by EHV-8 ( Figure 8 ). At the same time, lung tissues were collected at 7 dpi for grinding and TCID50 The results showed that tripterygium wilfordii can effectively reduce the replication of EHV-8 in the lungs ( Fig. 9 ).

[0061] In summary, the present invention takes traditional Chinese medicine technology as a starting point to find effective drug ingredients for resisting EHV-8 infection. By treating EHV-8 susceptible cells RK-13 ​​and NBL-6 with tripterygium wilfordii and conducting in vivo experiments, it was determined that tripterygium wilfordii had a significant inhibitory effect on the replication of EHV-8, further verifying that the drug had an inhibitory effect on multiple EHV-8 strains, and in-depth analysis found that tripterygium wilfordii could affect multiple replication stages of EHV-8, thereby proving that tripterygium wilfordii can be used to prepare drugs for preventing and treating EHV-8 infection.

[0062] The application of tripterygium wilfordii in the preparation of anti-EHV-8 drugs has the following advantages:

[0063] 1. Inhibit viral replication

[0064] The maximum safe mass concentration of tripterygium wilfordii at the susceptible cell level is 1 μmol / L, at which it can effectively inhibit EHV-8 replication.

[0065] In vitro, studies on EHV-8 susceptible cells RK-13 ​​and NBL-6 found that celastrol had a significant inhibitory effect on the replication of multiple strains of EHV-8 (such as SDLC66, SD2020113 and donkey / Shandong / 10 / 2021).

[0066] Analysis using qPCR and Western blot methods showed that tripterygium wilfordii could significantly inhibit EHV-8gD gene transcription and gD protein expression.

[0067] It was determined that tripterygium wilfordii can exert an antiviral effect in multiple stages of EHV-8 replication (such as pretreatment group, co-treatment group, post-infection incubation group, full-stage treatment group, etc.), but cannot directly inactivate EHV-8.

[0068] 2. Anti-EHV-8 effect in vivo

[0069] In the mouse model, tripterygium wilfordii can effectively reduce the replication level of EHV-8 in the lungs. For example, in an experiment on SPF BALB / c mice, different treatment groups were set up and it was found that the replication level of EHV-8 in the lungs of mice in the tripterygium wilfordii drug treatment group was significantly lower than that in the control group. Tripterygium wilfordii can significantly reduce the pathological damage caused by EHV-8. By formaldehyde fixation and histopathological examination of mouse lung tissue and tissue TCID 50 This has been confirmed by measurements and so on.

[0070] 3. Drug advantages

[0071] Tripterygium wilfordii can be used as an effective drug ingredient for the prevention and treatment of EHV-8 infection, giving full play to the advantages of natural compounds derived from plants with low toxicity and side effects.

[0072] Triptolide has anti-inflammatory effects and can reduce the damage of inflammatory responses to host cells. After being infected with EHV-8, host cells will release a variety of inflammatory factors. Triptolide can protect the host from the dual damage of virus and inflammation by inhibiting the release of inflammatory factors.

[0073] Tripterine has multiple pharmacological activities such as immunomodulation, anti-tumor, anti-fibrosis, and protection of cardiovascular and nervous systems, which provides it with more potential advantages as an anti-EHV-8 drug.

[0074] Triptolide has significant antiviral activity and has inhibitory effects on a variety of viruses (including hepatitis C virus, dengue virus, and new coronavirus). EHV-8 is a herpes virus that is highly pathogenic to equine animals. Therefore, triptolide can be used as a candidate drug for anti-EHV-8 drugs, which can protect the host from the virus by inhibiting the replication and spread of the virus and reducing the damage of the virus to the host cells.

[0075] Triptolide has anti-inflammatory effects and can reduce the damage of inflammatory response to host cells. After being infected with EHV-8, host cells will release a variety of inflammatory factors, leading to the occurrence of inflammatory response. Triptolide can reduce the damage of inflammatory response to host cells by inhibiting the release of inflammatory factors, thereby protecting the host from the dual damage of virus and inflammation.

[0076] In summary, the present invention relates to the use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs. The anti-EHV-8 study of tripterygium wilfordii in vitro and in vivo in mouse models revealed that tripterygium wilfordii can significantly inhibit the replication of EHV-8, can be used as an effective ingredient for the prevention and treatment of EHV-8 infection, and gives full play to the advantages of plant-derived natural compounds with low toxicity and side effects, providing valuable theoretical basis and reference for the clinical use of EHV-8 drugs, including antiviral and anti-inflammatory effects, which can protect the host from the dual damage of viruses and inflammation.

[0077] Studies have found that tripterygium wilfordii has significant anti-inflammatory, antioxidant and anti-tumor effects. The latest research has found that it has a good effect against the new coronavirus. EHV-8 is a herpes virus that is highly pathogenic to equine animals. The present invention confirms that tripterygium wilfordii significantly reduces the infection of EHV-8 in susceptible cells and mouse models.

[0078] Therefore, tripterygium wilfordii can be used as a candidate drug for anti-EHV-8 drugs, which can reduce the damage of viruses to host cells by inhibiting the replication and spread of viruses, thereby protecting the host from the invasion of viruses. The application of tripterygium wilfordii in the preparation of anti-EHV-8 drugs in vitro and in vivo in mouse models systematically carried out tripterygium wilfordii anti-EHV-8 research, revealing that tripterygium wilfordii can significantly inhibit the replication of EHV-8, can be used as an effective ingredient for preventing and treating EHV-8 infection, and give full play to the advantages of natural compounds from plant sources with low toxicity and side effects, providing valuable theoretical basis and reference for the clinical use of EHV-8.

[0079] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and descriptions are only preferred examples of the present invention and are not intended to limit the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. An application of tripterygium wilfordii in the preparation of an anti-EHV-8 drug, characterized in that: The invention includes the use of tripterygium wilfordii in preparing a medicine for inhibiting the replication of EHV-8.

2. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to claim 1, characterized in that: The invention also includes the application of the plant extract containing tripterygium wilfordii in the preparation of anti-EHV-8 medicine.

3. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to claim 1, characterized in that: It also includes the use of a traditional Chinese medicine compound containing tripterygium wilfordii in the preparation of anti-EHV-8 drugs.

4. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to any one of claims 1 to 3, characterized in that: The tripterygium wilfordii, the plant extract containing the tripterygium wilfordii and the traditional Chinese medicine compound containing the tripterygium wilfordii are used for preventing and treating respiratory diseases caused by EHV-8 infection of equine animals.

5. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to claim 4, characterized in that: The tripterygium wilfordii, the plant extract containing the tripterygium wilfordii and the traditional Chinese medicine compound containing the tripterygium wilfordii are used for preventing and treating abortion caused by EHV-8 infection in equine animals.

6. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to claim 4, characterized in that: The tripterygium wilfordii, the plant extract containing the tripterygium wilfordii and the traditional Chinese medicine compound containing the tripterygium wilfordii are used for treating viral encephalitis caused by EHV-8 infection.

7. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to claims 5 and 6, characterized in that: At the level of susceptible cells, the maximum safe mass concentration of the tripterygium wilfordii is 1 μmol / L.

8. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to any one of claims 1 to 3, characterized in that: The tripterygium wilfordii, the plant extract containing tripterygium wilfordii, and the traditional Chinese medicine compound containing tripterygium wilfordii all have inhibitory effects on multiple strains of EHV-8 (including SDLC66, SD2020113, and donkey / Shandong / 10 / 2021).

9. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to any one of claims 1 to 3, characterized in that: The tripterygium wilfordii, the plant extract containing the tripterygium wilfordii and the traditional Chinese medicine compound containing the tripterygium wilfordii all have inhibitory effects on multiple stages of EHV-8 replication.

10. The use of tripterygium wilfordii in the preparation of anti-EHV-8 drugs according to any one of claims 1 to 3, characterized in that: The tripterygium wilfordii, the plant extract containing the tripterygium wilfordii and the traditional Chinese medicine compound containing the tripterygium wilfordii are used for reducing lung damage in a mouse model caused by EHV-8 infection.

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