Application of tripterine in the preparation of anti-EHV-8 drugs

Abstract

The present invention relates to the technical field of traditional Chinese medicine production, specifically to the use of a traditional Chinese medicine, tripterine, in the preparation of an anti-EHV-8 drug, including the use of tripterine in the preparation of an anti-EHV-8 drug, the use of a plant extract containing tripterine in the preparation of an anti-EHV-8 drug, and the use of a traditional Chinese medicine compound containing tripterine in the preparation of an anti-EHV-8 drug. The present invention systematically conducts research on the inhibition of EHV-8 by tripterine in susceptible cells in vitro and in mouse models in vivo. The results show that tripterine can significantly inhibit the replication of EHV-8, can be used as an effective pharmaceutical ingredient for preventing and treating EHV-8 infection, and fully utilizes the advantage of natural compounds of plant origin with low toxicity and side effects, providing a 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.

Description

Technical Field

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

[0002] Previous studies have shown that equine herpesvirus type 8 (EHV-8) can cause rhinopneumonia in horses and donkeys, leading to respiratory distress and death in severe cases. Infection in pregnant horses and donkeys often manifests as a latent infection, with sudden miscarriage and, in some cases, neurological symptoms. This virus poses a serious health threat to equines. EHV-8 was first isolated from the nasal cavity of donkeys in Australia. Subsequently, Chinese researchers isolated the EHV-8 wh strain from horses for the first time in 2010. In 2020, Israeli researchers isolated the first wild-type EHV-8 from an adult male donkey.

[0003] EHV-8 is an alphaherpesvirus. Viral particles are approximately 150 nm in diameter, with a 100 nm diameter icosahedral nucleocapsid located within the particle. The mature virion is composed of a core, capsid, and envelope. The genome within the nucleocapsid is a linear scroll, while the lipoprotein envelope with its small glycoprotein spikes lies outside the nucleocapsid. Equine herpesvirus is an enveloped, double-stranded DNA virus with a genome of approximately 150 kb, containing 80 open reading frames and encoding 76 proteins.

[0004] With the rapid development of the horse and donkey industries in recent years, epidemics have been increasing on large-scale farms, particularly high abortion rates and frequent respiratory illnesses, seriously hindering the sustainable development of the industry. Our team's preliminary research has identified EHV-8 as the primary pathogen causing abortions, respiratory illnesses, and viral encephalitis in donkeys. We have isolated multiple strains of wild-type EHV-8 from the lungs, placentas, and brain tissue of dead donkeys. Currently, there are no effective vaccines or medications for this virus, and a cost-effective 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 Modern pharmacological studies have shown that this drug exhibits multiple pharmacological activities, including immunomodulatory, anti-inflammatory, anti-tumor, anti-fibrotic, and cardiovascular and nervous system protection, with a good in vivo safety profile. In recent years, it has garnered widespread attention in anti-inflammatory and anti-tumor research due to its low adverse reaction profile, high efficacy, broad spectrum, multi-target activity, and low resistance profile. However, there are currently no reports on the use of tripterygium wilfordii in the prevention and treatment of EHV-8 infection. Summary of the Invention

[0006] The purpose of the present invention is to provide a use of tripterine 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 tripterine in preparing an anti-EHV-8 drug, including the application of tripterine in preparing a drug for inhibiting EHV-8 replication.

[0009] As a preference, the invention also includes the use of a plant extract containing tripterine in the preparation of an anti-EHV-8 drug.

[0010] As a preference, the invention also includes the use of a traditional Chinese medicine compound containing tripterine 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 tripterine, the plant extract containing tripterine and the traditional Chinese medicine compound containing tripterine 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 the tripterygium wilfordii is 1µmol / L.

[0015] Preferably, the tripterine, the plant extract containing tripterine, and the traditional Chinese medicine compound containing tripterine 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 concentration of tripterygium wilfordii in susceptible cells (RK-13 and NBL-6) is 1µmol / L, at which it can effectively inhibit viral replication.

[0021] In vitro studies on EHV-8 susceptible cells, RK-13 and NBL-6, revealed that tripterygium wilfordii significantly inhibited the replication of EHV-8 SDLC66. Quantitative polymerase chain reaction (qPCR) and Western blot analyses demonstrated that tripterygium wilfordii significantly inhibited EHV-8 gD gene transcription and gD protein expression.

[0022] In vitro, the IFA method was used to confirm that tripterygium wilfordii 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] qPCR and Western blot methods were used to determine that tripterygium wilfordii could 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 could not 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 in the tripterygium wilfordii drug-treated group was significantly lower than that in the control group.

[0026] 3. Drug advantages

[0027] Triptolide can be used as an effective drug ingredient for preventing and treating EHV-8 infection, giving full play to the advantages of natural compounds derived from plants with few toxic and side effects.

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

[0029] Triptolide 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 was systematically studied in vitro and in vivo in mouse models to investigate the anti-EHV-8 effect of tripterygium wilfordii, revealing that tripterygium wilfordii can significantly inhibit the replication of EHV-8, and can be used as an effective ingredient in drugs for preventing and treating EHV-8 infection. It also gives full play to the advantages of natural compounds of plant origin with low toxicity and side effects, and provides a 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 virus 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 are shown in FIG.

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

[0035] Figure 4 The results of indirect immunofluorescence (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 in 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 cannot directly inactivate EHV-8;

[0039] Figure 8 This is a HE staining image showing that the tripterygium wilfordii of the present invention reduces the damage caused by EHV-8 in the mouse lungs;

[0040] Figure 9 This is a graph showing the results of the present invention's tripterygium wilfordii reducing the replication level of EHV-8 in mouse lungs. 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 embodiments described 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 making creative efforts 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 into 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 dilutions were added to each well. At the same time, a DMSO negative control and a 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 h, 10 μL of CCK-8 solution was added to each well and cultured for another 2 h. The absorbance value at OD 450nm was read with the help of a microplate reader, and the effect of tripterygium wilfordii on the activity of RK-13 and NBL-6 cells was calculated using the following formulas, respectively, to evaluate the maximum safe mass concentration of tripterygium wilfordii (the results are shown in the figure). 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 containing 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 in the results, different concentrations of tripterygium wilfordii had no significant effect on the viability of RK-13 and NBL-6 cells, and the maximum safe concentration was 1 μM.

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

[0050] 1. Analysis of EHV-8 gD Gene Expression by qPCR and Western Blot

[0051] RK-13 and NBL-6 cells were seeded into 6-well cell plates (2 × 105 Cells were pretreated with different concentrations of tripterine (0 μM, 0.25 μM, 0.5 μM, and 1 μM) for 2 h when the cells were confluent to 80%. After inoculation with EHV-8 SDLC66 (0.1 MOI) for 1 h, the virus solution was discarded and cell maintenance medium containing tripterine was added. A DMSO-treated group and a normal cell untreated group were set as control groups. 24 h after virus infection, the cell supernatant was collected, and the total cell DNA was extracted using a kit. The changes in the EHV-8 gD virus copy number were determined by qPCR. The results are shown in the figure. Figure 3 shown.

[0052] RK-13 and NBL-6 cells were seeded in 6-well plates (2 × 10 5 Cells were cultured for 2 h after 80% confluence, and different concentrations of tripterine (0 μM, 0.25 μM, 0.5 μM, and 1 μM) were added. EHV-8 SDLC66 (0.1 MOI) was inoculated for 1 h, and the virus solution was discarded. Cell maintenance medium containing tripterine was added. DMSO-treated and normal cell untreated groups were set as control groups. Cells were collected 24 h after infection, and the changes in EHV-8 gD protein expression levels were analyzed by Western blot. Figure 3 As shown in the results, tripterine 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 seeded into 12-well cell culture plates. When the cells reached 80% confluence, they were pretreated with different concentrations of tripterygium wilfordii (0.25 μM, 0.5 μM, and 1 μM). The DMSO-treated group served as a negative control. After 2 h, the drug solution was discarded. The cells were infected with EHV-8SDLC66, EHV-8 SD2020113, and EHV-8 donkey / Shandong / 10 / 2021 for 1 h, respectively. The supernatant containing the drug and virus was discarded and replaced with 3% FBS DMEM. After 48 h of infection, the cells were fixed with 75% ethanol. Mouse anti-EHV-8 positive serum was used as the primary antibody, and CY3-labeled goat anti-mouse fluorescent antibody was used as the secondary antibody. The effect of tripterygium wilfordii on EHV-8 replication was evaluated by indirect immunofluorescence assay (IFA). The results are shown in Figure 2. Figure 4 shown.

[0055] 3. Determine the Effect of Tripterygium Wilfordii on the EHV-8 Replication Cycle

[0056] To further explore the mechanism of tripterygium wilfordii affecting EHV-8 infection, RK-13 and NBL-6 cells were seeded 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 group incubated with EHV-8 after infection (Post), and the celastrol full-stage treatment group (All stages-treatment) were set up respectively (see Figure 5 ), EHV-8 SDLC66 was inoculated (MOI = 0.1), and cells were collected 24 hours later to detect viral copy number by qPCR and to evaluate gD protein expression by Western blot. Figure 6 This suggests that tripterygium wilfordii exerts antiviral effects at multiple stages of EHV-8 replication.

[0057] 4. Determine whether tripterygium wilfordii has the activity to inactivate 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 Cells were cultured overnight. Triptolide (1 μM) was incubated with different doses of EHV-8 SDLC66 (0.1 MOI, 0.5 MOI, and 1 MOI) at 37°C for 2 h. After 1 h of infection, the cells were replaced with 3% FBS MEM for 24 h. The cell supernatants were collected and the viral copy number was detected by qPCR. The results are shown in Figure 2. Figure 7 This suggests that tripterygium wilfordii cannot directly inactivate EHV-8.

[0059] 5. In vivo anti-EHV-8 infection study of tripterygium wilfordii

[0060] Twelve SPF-grade BALB / c mice were randomly divided into four groups, with three mice in each group. A negative control group (Mock); a DMSO-treated control group + EHV-8; a 2 mg / kg tripterygium wilfordii group + EHV-8; and a 4 mg / kg tripterygium wilfordii group + EHV-8 were set up. One day before infection, mice in the celastrol-treated group received an intraperitoneal injection of the corresponding dose of the drug, while mice in the EHV-8-infected control group received an equal dose of DMSO solution. Except for the negative control group, mice in each group were intranasally infected with 100 μL (1 × 10 5 PFU / mouse) EHV-8SDLC66, were administered once at fixed time points on the first day after infection (i.e., 1 dpi) and the third day after infection (i.e., 3 dpi), and their body weight changes and status were detected every day. The mice were killed by cervical dislocation on the 7th day after virus infection (i.e., 7 dpi), and the lungs were collected for formaldehyde fixation and histopathological examination. The results showed that tripterine 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 TCID 50 The results showed that tripterygium wilfordii can effectively reduce the replication of EHV-8 in the lungs ( Figure 9 ).

[0061] In summary, the present invention uses traditional Chinese medicine technology as a starting point to identify effective pharmaceutical ingredients for combating 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 significantly inhibited EHV-8 replication. This inhibitory effect against multiple EHV-8 strains was further verified, and in-depth analysis revealed that tripterygium wilfordii affected multiple stages of EHV-8 replication, demonstrating that tripterygium wilfordii can be used to prepare a drug 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 concentration of tripterygium wilfordii at the susceptible cell level is 1µmol / 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] qPCR and Western blot analysis showed that tripterygium wilfordii could significantly inhibit EHV-8 gD gene transcription and gD protein expression.

[0067] It was determined that tripterygium wilfordii could 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 could not directly inactivate EHV-8.

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

[0069] In a 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 treated with tripterygium wilfordii was significantly lower than that of 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 other tests.

[0070] 3. Drug advantages

[0071] Triptolide can be used as an effective drug ingredient for preventing and treating EHV-8 infection, giving full play to the advantages of natural compounds derived from plants with few toxic and side effects.

[0072] Celastrol has anti-inflammatory properties and can reduce the damage caused by inflammatory responses to host cells. After infection with EHV-8, host cells release a variety of inflammatory factors. Celastrol can protect the host from the dual damage of the virus and inflammation by inhibiting the release of inflammatory factors.

[0073] Triptolide 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] Celastrol has significant antiviral activity, inhibiting a variety of viruses, including hepatitis C virus, dengue virus, and novel coronavirus. EHV-8 is a herpesvirus that is highly pathogenic to equines. Therefore, celastrol could be a candidate anti-EHV-8 drug, potentially protecting the host from viral damage by inhibiting viral replication and spread and mitigating viral damage to host cells.

[0075] Celastrol has anti-inflammatory properties and can reduce the damage caused by inflammatory responses to host cells. After infection with EHV-8, host cells release a variety of inflammatory factors, leading to an inflammatory response. Celastrol can inhibit the release of inflammatory factors and reduce the damage caused by inflammatory responses to host cells, thereby protecting the host from the dual damage of the virus and inflammation.

[0076] In summary, the present invention relates to the use of tripterine in the preparation of anti-EHV-8 drugs. Systematic in vitro and in vivo studies of tripterine against EHV-8 revealed that tripterine can significantly inhibit EHV-8 replication, making it a useful active ingredient in drugs for preventing and treating EHV-8 infection. This study leverages the advantages of natural plant-derived compounds with minimal toxicity and side effects, providing a valuable theoretical basis and reference for the clinical use of EHV-8 drugs. The drug exhibits antiviral and anti-inflammatory effects, protecting the host from both viral and inflammatory damage.

[0077] Research has shown that tripterine has significant anti-inflammatory, antioxidant, and anti-tumor effects. Recent studies have shown that it has excellent efficacy against the novel coronavirus. EHV-8 is a herpes virus that is highly pathogenic to equines. This study demonstrates that tripterine significantly reduces EHV-8 infection in susceptible cells and mouse models.

[0078] Therefore, tripterine can be used as a candidate drug for anti-EHV-8 drugs, which can reduce the damage of the virus to host cells by inhibiting the replication and spread of the virus, thereby protecting the host from the invasion of the virus. The application of tripterine in the preparation of anti-EHV-8 drugs of the present invention is to carry out tripterine anti-EHV-8 studies in vitro and in vivo in mouse models, revealing that tripterine 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 low toxicity and side effects, providing valuable theoretical basis and reference for the clinical use of EHV-8 drugs.

[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 to the above embodiments. The above embodiments and descriptions are merely preferred examples of the present invention and are not intended to limit the present invention. Various changes and improvements may be made to the present invention without departing from the spirit and scope of the present invention. Such changes and improvements fall within the scope of the present invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. An application of tripterine in the preparation of an anti-EHV-8 drug, characterized in that: The invention also includes the use of tripterine in preparing medicines for inhibiting EHV-8 replication.

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

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

4. The use of tripterine according to any one of claims 1 to 3 in the preparation of anti-EHV-8 drugs, 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 in equine animals.

5. The use of tripterine according to claim 4 in the preparation of anti-EHV-8 drugs, 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 tripterine according to claim 4 in the preparation of anti-EHV-8 drugs, 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 of equine animals.

7. The use of tripterine according to claim 5 or 6 in the preparation of an anti-EHV-8 drug, characterized in that: At the susceptible cell level, the maximum safe concentration of the tripterygium wilfordii is 1µmol / L.

8. The use of tripterine according to any one of claims 1 to 3 in the preparation of anti-EHV-8 drugs, 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 the SDLC66, SD2020113 and donkey / Shandong / 10 / 2021 strains of EHV-8.

9. The use of tripterine according to any one of claims 1 to 3 in the preparation of anti-EHV-8 drugs, 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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