Application of miR-145-5P in preparation of medicine for preventing and / or treating pseudorabies virus infection related diseases

Abstract

The invention relates to application of miR-145-5P in preparation of a medicine for preventing and / or treating PRV (pseudorabies virus) infection related diseases, and belongs to the field of biological medicines. The miR-145-5p is found to be a key differential expression molecule in the PRV infected piglet lung and colon tissue exosome through high-throughput sequencing for the first time. In-vitro cell experiments prove that up-regulation of the expression of the miR-145-5p can significantly play an antiviral role through double mechanisms of inhibiting PRV virus replication and promoting apoptosis of infected cells, and research results of the invention provide a brand new target and strategy for prevention and treatment of PRV.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically, it relates to the use of miIR-145-5P in the preparation of drugs for the prevention and / or treatment of diseases related to pseudorabies virus (PRV) infection. Background Technology

[0002] Pseudorabies virus (PRV), also known as porcine herpesvirus A, is the pathogen that causes porcine pseudorabies (Aujeszky's disease). This disease causes significant economic losses to the global pig industry, clinically manifesting as high mortality rates in piglets, respiratory symptoms in finishing pigs, and reproductive disorders in breeding pigs. Notably, PRV has a broad host range, infecting various mammals, including humans, making it a potential and highly concerning zoonotic pathogen.

[0003] Currently, the prevention and control of PRV mainly relies on vaccination (such as live attenuated vaccines like Bartha-K61) and biosafety measures. However, since 2011, the emergence and spread of PRV variants have led to a decline in the protective efficacy of traditional vaccines against circulating strains, and even frequent cases of immunization failure, making the development of novel antiviral strategies particularly urgent. Existing antiviral drugs (such as nucleoside analogs) have limited efficacy in the treatment of PRV and may have side effects; therefore, finding new drug targets is a current research focus.

[0004] In recent years, the role of exosomes as key mediators of intercellular communication in viral infection has been gradually revealed. Exosomes are nanoscale extracellular vesicles, approximately 30-200 nanometers in diameter, secreted by cells. Their biogenesis depends on the endosome system and they carry specific marker proteins (such as CD63 and TSG101). Exosomes can carry functional nucleic acid molecules such as microRNAs (miRNAs) and transfer them between donor and recipient cells, thereby precisely regulating the physiological state of the recipient cells, including viral replication and the host's immune response. For example, studies have reported that certain human viruses (such as enterovirus 71 EV-A71 and hepatitis C virus HCV) can use exosomal miRNAs to evade host immunity. However, these studies have mostly focused on human viruses or using cell line models. Whether and how infection with important animal pathogens such as PRV regulates host tissue-derived exosomal miRNAs remains unknown.

[0005] miR-145-5p is a conserved miRNA in organisms, and its functions in tumorigenesis and angiogenesis have been reported. Although some sporadic studies suggest that miR-145-5p may be related to certain viral infections, its specific role and expression patterns in PRV infection have not yet been systematically studied and definitively concluded. Summary of the Invention

[0006] To address the gaps in the existing technology, the present invention aims to provide a new use for miR-145-5p and related products, particularly its application in combating pseudorabies virus (PRV) infection. Through in vitro and in vivo experiments, this invention reveals and verifies for the first time the key negative regulatory role of miR-145-5p in PRV infection and its molecular mechanism of influencing PRV replication through the apoptosis pathway.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solution: The first objective of this invention is to provide the use of miR-145-5p in the preparation of a medicament for the prevention and / or treatment of diseases related to pseudorabies virus (PRV) infection.

[0008] Furthermore, the miR-145-5p is at least one of miR-145-5p, miR-145-5p analogue, or miR-145-5p agonist.

[0009] A second objective of this invention is to provide the use of a detection reagent marked with miR-145-5p as a diagnostic reagent or kit for pseudorabies virus (PRV) infection.

[0010] A third objective of this invention is to provide a reagent / kit that uses miR-145-5p as a detection marker.

[0011] Furthermore, the reagent or kit contains primers and / or probes for the specific detection of miR-145-5p.

[0012] A fourth objective of the present invention is to provide a pharmaceutical composition for the prevention and / or treatment of pseudorabies virus infection, the composition comprising an effective amount of miR-145-5p mimic and a pharmaceutically acceptable carrier.

[0013] Furthermore, pharmaceutically acceptable carriers include liposomes, exosomes, or biodegradable nanoparticles.

[0014] The present invention also provides a method for inhibiting pseudorabies virus replication in vitro, characterized in that an effective amount of miR-145-5p mimic is introduced into cells infected or potentially infected with PRV.

[0015] The beneficial effects of this invention are: (1) This invention is the first to identify the differential expression of miR-145-5p in PRV-infected tissue-derived exosomes: High-throughput sequencing and RT-qPCR verification revealed that, compared with the normal control group, the expression level of miR-145-5p in lung tissue exosomes of PRV-infected piglets was significantly downregulated, while the expression level of miR-145-5p in colon tissue exosomes was significantly upregulated. This indicates that miR-145-5p is a key response factor in the PRV infection process, and its response pattern is tissue-specific.

[0016] (2) Through research, this invention found that miR-145-5p expression changed significantly in PRV-infected cell models. In PRV-infected 3D4 / 21 (porcine alveolar macrophages) and IPEC-J2 (porcine small intestinal epithelial cells), the relative expression level of intracellular miR-145-5p was significantly increased, which confirmed for the first time the direct association between miR-145-5p and PRV infection.

[0017] (3) The study of this invention confirms that the expression level of miR-145-5p is negatively correlated with PRV virus replication, and upregulating its expression can directly inhibit virus replication.

[0018] (4) This invention demonstrates for the first time that miR-145-5p limits viral amplification and spread by promoting apoptosis in PRV-infected cells.

[0019] (5) This invention expands the new perspective of molecular regulation of the "lung-gut axis" in PRV infection: This invention found that miR-145-5p can play a consistent antiviral and pro-apoptotic role in both lung-derived 3D4 / 21 cells and gut-derived IPEC-J2 cells, suggesting that miR-145-5p may be a key molecule connecting the lung and gut and coordinating the anti-PRV infection response, providing a new direction for understanding the systemic pathogenic mechanism of PRV and developing broad-spectrum antiviral strategies. Attached Figure Description

[0020] Figure 1 The body temperature of piglets in the PRV-infected group and NC group in Example 1 of this invention; Figure 2 These are microscopic pathological images of the lungs and colon tissues of piglets in the PRV-infected group and NC group in Example 1 of this invention; Figure 3 This refers to the pathogen PCR detection results in Example 1 of the present invention; Figure 4 This is a transmission electron microscope image of exosomes from Embodiment 1 of the present invention; Figure 5 This is an NTA diagram of exosomes from Embodiment 1 of the present invention; Figure 6 This is a WB image of exosomes from Embodiment 1 of the present invention; Figure 7 These are the volcano diagram and KEGG functional enrichment diagram of differential miRNA expression in exosomes of the lung and colon tissues of the PRV and NC groups in Example 2 of this invention; Figure 8 This is a heatmap of differential miRNA expression in exosomes of colon tissue in the PRV and NC groups in Example 2 of this invention; Figure 9 The differentially expressed miRNAs in colon tissue exosomes of the PRV group and NC group in Example 2 of this invention; Figure 10 This refers to the relative expression levels of miR-145-5p in IPEC and 3D4 / 21 cells in the PRV group and NC group in Example 3 of this invention. Figure 11 This is a statistical graph showing the transfection efficiency of IPEC and 3D4 / 21 cells transfected with different concentrations of miR-145-5p inhibitors and mimics. Figure 12 The PRV viral load in IPEC and 3D4 / 21 cells transfected with 50 nM concentration of miR-145-5p inhibitor and mimic; Figure 13 This is the result of flow cytometry analysis (apoptosis rate of IPEC cells transfected with miR-145-5p mimics). Figure 14 This is the result of flow cytometry analysis (apoptosis rate of IPEC cells transfected with miR-145-5p inhibitor). Detailed Implementation

[0021] To make the objectives, technical solutions, and beneficial effects of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are all within the scope of protection of this invention.

[0022] The main reagents and materials used in the following examples are: Cells: porcine alveolar macrophages 3D4 / 21 (purchased from Beina Biotechnology, catalog number: BNCC338298); porcine small intestinal epithelial cells IPEC-J2 (preserved in the laboratory).

[0023] Strain: PRV-XD-F3 strain.

[0024] Experimental animals: Six healthy male Zhaotong Wujin pigs, aged 50±3 days and weighing 10±1 kg.

[0025] Main reagents: Tissue exosome extraction kit (Umibio, UR52161); miRNA extraction kit (TransGold, ER601-01-V2); TransScript® miRNA First-Strand cDNA Synthesis SuperMix (TransGold, AT351-01); SYBR Green Real-time PCR Master Mix (Takara, RR820A); Lipofectamine 3000 transfection reagent; fetal bovine serum; DMEM medium (Gibco); Anti-TSG101 antibody (Abcam, ab125011); CD63 antibody (Affinity, AF5117).

[0026] miR-145-5p mimic and inhibitor: Synthesized by Shanghai Sangon Biotech Co., Ltd., its sequence follows the mature porcine miR-145-5p sequence in the miRBase database. The mimic is labeled with FAM fluorescence to facilitate observation of transfection efficiency.

[0027] Example 1: Construction of a PRV-infected piglet model and isolation and identification of tissue exosomes 1.1 Animal Model Construction Six piglets were randomly divided into a PRV-infected group (PRV group) and a negative control group (NC group), with three piglets in each group. On day 0 of the experiment, each piglet in the PRV group was injected intramuscularly with 1 mL of PRV-XD-F3 virus solution (TCID50 10⁻⁶.13), while the NC group was injected with an equal volume of sterile PBS. Clinical symptoms (temperature, diarrhea, cough, etc.) and mortality were observed and recorded for seven consecutive days. Results (e.g., ...) were recorded. Figure 1 The results showed that piglets in the PRV group developed fever on day 1 after viral challenge, followed by diarrhea and coughing on day 2. Piglets in the NC group showed no obvious clinical symptoms.

[0028] 1.2 Sample Collection and Pathological Examination Seven days after viral challenge, all piglets underwent necropsy. Gross pathological changes in the lungs and colon were observed, and tissue samples were collected. Some tissues were fixed in 4% paraformaldehyde, dehydrated, cleared, embedded in paraffin, sectioned, stained with H&E, and observed under a light microscope for microscopic pathological changes. The necropsy results are attached. Figure 2The results showed that in the PRV group, obvious needle-like hemorrhages were visible in the lungs of piglets, and the colon tissue showed varying degrees of edema, with significantly enlarged mesenteric lymph nodes; while no obvious gross pathological changes were observed in the NC group. HE staining results showed that the lung tissue structure of the NC group was intact, the alveoli were regularly arranged, and the alveolar walls were thinner; in contrast, the alveolar walls of the PRV group were significantly thickened, with increased infiltration of inflammatory cells around blood vessels and bronchi, and widening of the lung interstitium. In the colon tissue, the mucosal epithelium of the NC group was intact and regularly arranged; the colonic mucosal epithelium of the PRV group showed varying degrees of shedding and damage, and widening of the interstitial spaces, indicating damage to the intestinal tissue structure.

[0029] 1.3 Pathogen detection To confirm model specificity, PCR was used to detect PRV, classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), and porcine circovirus type 2 (PCV-2) in lung and colon tissue samples. Primer sequences are shown in Table 1. Results (attached) Figure 3 The results showed that only the PRV test was positive, ruling out interference from other common pathogens.

[0030] Table 1 Primer Sequences 1.4 Isolation and Identification of Tissue Exosomes Lung and colon tissues that tested positive for PRV and showed significant pathological damage were collected and processed according to the instructions of the tissue exosome extraction kit.

[0031] Tissue mincing and digestion: Mince approximately 0.1g of tissue, add 1 mL of Solution A2, incubate at 37°C and 80 rpm on a shaker for 20 minutes, centrifuge at 10000 rpm for 10 minutes, and collect the supernatant.

[0032] Exosome extraction and purification: Add 1 / 4 volume of Solution B2 to the supernatant, vortex to mix, and incubate at 4°C for 20 minutes. Centrifuge at 10,000 rpm for 20 minutes, discard the supernatant, and resuspend the precipitate in 200 μL PBS. Transfer the resuspended solution to a purification column, centrifuge at 4°C and 6,000 rpm for 10 minutes, and collect the eluent as the purified exosomes.

[0033] Exosome identification: Transmission electron microscopy: 10 μL of exosome suspension was dropped onto a copper grid, negatively stained with 1% phosphotungstic acid, and observed. Results of transmission electron microscopy observation ( Figure 4 The results showed that both groups of exosomes were morphologically intact, exhibiting typical round or spherical structures, varying in size, with clear boundaries and a distinct double membrane structure, consistent with the morphological characteristics of exosomes.

[0034] Nanoparticle tracking and analysis: Instruments ( Figure 5The results showed that the main particle size peak was 126.1 nm, which is consistent with the characteristics of exosomes.

[0035] Western Blot ( Figure 6 The TSG101 and CD63 protein bands were 44 kDa and 25 kDa, respectively, which are within the molecular size range of TSG101 and CD63 proteins, indicating that both exosome surface marker proteins TSG10 and CD63 are expressed.

[0036] Example 2: Exosomal miRNA sequencing and screening verification of miR-145-5p 2.1 Total RNA extraction and sequencing Exosome samples from the lungs and colon (three biological replicates each) of the NC and PRV groups were collected, and total RNA was extracted using the Trizol LS method. Small RNA libraries were constructed using the GenSeq® Small RNA Library Prep Kit by Shanghai Sangon Biotech Co., Ltd., and high-throughput sequencing was performed on the Illumina sequencing platform.

[0037] 2.2 Bioinformatics Analysis After quality control and alignment of sequencing data, novel miRNAs were predicted using miRDeep2 software, and known miRNAs were annotated by comparison with the miRBasev21.0 database. Normalization was performed using TPM values, and differentially expressed miRNAs were screened based on |Fold Change| > 2 and padj < 0.05. Target genes were predicted using TargetScan and miRanda software, and GO function and KEGG pathway enrichment analyses were performed. Figure 7 and Figure 8 . Figure 7 In the diagram, A shows the lungs of the PRV and NC groups, B shows the colon of the PRV and NC groups, and C and D show the differential miRNA KEGG function maps (first 20, C shows the lungs of the PRV and NC groups, and D shows the colon of the PRV and NC groups). Figure 8 In the diagram, Figure A shows the lungs of the PRV and NC groups, and Figure B shows the colons of the PRV and NC groups.

[0038] The results showed that compared with the control group, the number of upregulated miRNAs in the lungs was 49, and the number downregulated was 46. Figure 7 A); 50 colonic miRNAs were upregulated, and 63 were downregulated. Figure 7 B), and differential miRNA KEGG pathway analysis showed that differentially expressed miRNAs between PRV and NC were significantly enriched in the apoptosis signaling pathway. Figure 7 C,D) Table 2. Differential miRNAs between PRV-lung and NC-lung (top 30) Table 3. Differential miRNAs between PRV-colon and NC-colon (top 30) KEGG enrichment analysis showed that differentially expressed exosomal miRNAs (including miR-145-5p) under PRV infection were significantly enriched in the apoptosis signaling pathway, providing a theoretical basis for the above functional experiments.

[0039] 2.3 RT-qPCR Validation Differentially expressed miRNAs such as miR-145-5p (as shown in the table below) were screened and validated by RT-qPCR using the SYBR Green method with U6 as an internal control. The reaction mixture (20 μL) consisted of 10 μL SYBR Green Master Mix, 0.4 μL each of forward and reverse primers, 2 μL cDNA, and 7.2 μL ddH2O. The reaction program was 95℃ for 30 s; 95℃ for 5 s, 60℃ for 30 s, for 40 cycles. Relative expression levels were calculated using the 2^(-ΔΔCt) method. The expression levels of various miRNAs in lung and colon tissues of the NC and PRV groups were compared and analyzed. The results (…) Figure 9The study showed that PRV infection induces significant differential regulatory patterns of different miRNAs in lung and colon tissues. In the expression analysis of miR-145-5p, compared with the NC group, the expression level of miR-145-5p in the lung tissue of the PRV group was significantly decreased (P < 0.05), while it was significantly increased in the colon tissue (P < 0.01), suggesting that PRV infection has significant tissue specificity in regulating miR-145-5p. miR-24-3p was significantly downregulated in the lung tissue of the PRV group (P < 0.01), while it was significantly upregulated in the colon tissue (P < 0.05), showing a similar lung-intestinal differential expression trend as miR-145-5p. For miR-151-5p, PRV infection led to a significant decrease in its expression in lung tissue (P < 0.05), while it was significantly increased in colon tissue (P < 0.01), suggesting that this miRNA may be involved in the regulation of different tissue responses during PRV infection. miR-191 was significantly downregulated in lung tissue of the PRV group (P < 0.05) but significantly upregulated in colon tissue (P < 0.01), further indicating that PRV infection can induce tissue-specific changes in miRNA expression. In let-7c expression analysis, let-7c expression was significantly decreased in lung tissue of the PRV group (P < 0.01) but significantly increased in colon tissue (P < 0.05), suggesting it may be involved in the regulation of PRV infection-related inflammation or tissue damage. let-7a was significantly downregulated in both lung and colon tissues of the PRV group (P < 0.05), showing a relatively consistent inhibitory trend of this miRNA under PRV infection conditions. Furthermore, miR-126-3p was significantly upregulated in both lung and colon tissues of the PRV group (P < 0.01), suggesting it may play a promoting or responsive role in PRV infection-related pathological processes. miR-143-3p was significantly upregulated in lung tissue of the PRV group (P < 0.01), but significantly downregulated in colon tissue (P < 0.01), further demonstrating the tissue specificity of PRV infection in miRNA regulation. Overall, the expression trend of miR-145-5p was consistent with the sequencing results.

[0040] Table 5 miRNA primer sequences Example 3: Functional validation of miR-145-5p in a cell model 3.1 Cell infection model and expression detection 3D4 / 21 and IPEC-J2 cells were cultured in complete medium containing 10% fetal bovine serum. When cell confluence reached 80%, the cells were injected with 100-fold TCID50. 50 Cells were infected with PRV. Twenty-four hours after infection, cells were collected, total RNA was extracted, and the expression level of miR-145-5p was detected by RT-qPCR. Results ( Figure 10 The results showed that, compared with the uninfected control group, the expression level of miR-145-5p in the PRV-infected group was significantly increased (*p* < 0.01), further confirming the direct association between miR-145-5p and PRV infection.

[0041] 3.2 Optimization of Transfection Conditions Cells were seeded in 6-well plates and transfected when confluence reached 60%-70%. The miR-145-5p mimic or inhibitor was diluted to different concentration gradients (10, 30, 50, 100 nM) with 125 μL Opti-MEM, and simultaneously, 5 μL Lipofectamine 3000 was diluted with 125 μL Opti-MEM. After incubation at room temperature for 5 minutes, the mixture was mixed and incubated for another 20 minutes to form a transfection complex. The complex was added to the wells, and after 4-6 hours of transfection, the medium was replaced with complete medium (100 mL of medium supplemented with 90 mL DMEM, 10 mL of fetal bovine serum, and 1 mL of penicillin-dextrin antibiotics, stored at 4 °C). 24 hours after transfection, the FAM fluorescence intensity was observed under a fluorescence microscope, and the expression changes of miR-145-5p or its target gene TLR4 were detected by qPCR. Figure 11 ), and determined that **50 nM** was the optimal transfection concentration.

[0042] 3.3 Viral load detection Experimental groups: 1) Blank control; 2) Mimic negative control; 3) Inhibitor negative control; 4) miR-145-5p mimic transfection group; 5) miR-145-5p inhibitor transfection group; 6) PRV infection group; 7) Mimic transfection + PRV infection group; 8) Inhibitor transfection + PRV infection group.

[0043] Twenty-four hours after transfection, groups 6, 7, and 8 were challenged with PRV. Twenty-four hours after infection, cells were collected, and genomic DNA was extracted. Absolute quantitative qPCR was performed using a probe targeting the PRV gE gene. The results were analyzed according to the standard curve (y = -2.3948x +37.091, R0). 2 =0.993) calculate the virus copy number. Result ( Figure 12The results showed that in 3D4 / 21 cells, compared with the PRV infection group, viral load was significantly reduced after transfection with miR-145-5p mimic (p < 0.0001); while viral load was significantly increased after transfection with miR-145-5p inhibitor (p < 0.0001).

[0044] A completely consistent trend was observed in IPEC-J2 cells: miR-145-5p mimics significantly reduced viral load (p < 0.0001), while inhibitors significantly increased it (p < 0.0001).

[0045] These results demonstrate that the expression level of miR-145-5p is negatively correlated with PRV viral replication, and upregulating its expression can directly inhibit viral replication.

[0046] 3.4 Apoptosis rate detection Cell processing and grouping were the same as in Example 3.3. The apoptosis rate was detected by flow cytometry using Annexin V-FITC / PI double staining. Specific steps: Cells were collected by trypsin digestion, washed with PBS, and resuspended in 100 μL of 1× Binding Buffer. 5 μL of Annexin V-FITC and 5 μL of PI staining solution were added, and the cells were incubated at room temperature in the dark for 15 minutes. Then, 400 μL of Binding Buffer was added, and the cells were analyzed within 1 hour. Flow cytometry analysis results ( Figure 13 and Figure 14 The results showed that, compared with the PRV group, the apoptosis rate was significantly increased in the miR-145-5p mimic transfection + PRV group (*p* < 0.001 ~ *p* < 0.0001), while the apoptosis rate was extremely significantly decreased in the inhibitor transfection + PRV group (*p* < 0.0001). In 3D4 / 21 cells, compared with the PRV infection group, the apoptosis rate was significantly increased after transfection with miR-145-5p mimic (p < 0.001), while the apoptosis rate was extremely significantly decreased after transfection with its inhibitor (p < 0.0001). In IPEC-J2 cells, the miR-145-5p mimic significantly increased the apoptosis rate (p < 0.0001), while the inhibitor significantly decreased it (p < 0.0001).

[0047] It should also be noted that: The miR-145-5p mimic of this invention is a chemically synthesized double-stranded RNA molecule with the same sequence as endogenous mature miR-145-5p. Its function is to mimic and enhance the activity of endogenous miR-145-5p within cells. This mimic can be obtained through commercially available solid-phase synthesis techniques and can be chemically modified (such as 2'-methoxy modification, cholesterol modification, or FAM fluorescent labeling) to enhance its stability or facilitate tracking.

[0048] The "prevention and / or treatment of pseudorabies virus infection" described in this invention includes, but is not limited to, any or all of the following aspects: inhibiting PRV replication in host cells or individuals; reducing viral load caused by PRV infection; alleviating clinical symptoms caused by PRV infection (such as fever, diarrhea, respiratory and neurological symptoms); improving histopathological damage caused by PRV infection (such as pulmonary hemorrhage, colonic edema, inflammatory cell infiltration); and clearing infected cells by regulating apoptosis pathways.

[0049] In another specific embodiment of the present invention, a method for preparing a miR-145-5p mimic is provided. The mimic is obtained by chemical synthesis and purification by a professional bioengineering company (such as Shanghai Sangon Biotech Co., Ltd.) based on the mature porcine miR-145-5p sequence (serial number: MIMAT0002154) provided by the miRBase database.

[0050] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. The use of miR-145-5p in the preparation of drugs for the prevention and / or treatment of diseases related to pseudorabies virus (PRV) infection.

2. The application according to claim 1, wherein the miR-145-5p is at least one of miR-145-5p, miR-145-5p analogue, or miR-145-5p agonist.

3. Application of detection reagents using miR-145-5p as markers in the preparation of diagnostic reagents or kits for pseudorabies virus (PRV) infection.

4. Reagents / kits using miR-145-5p as the detection marker.

5. The reagent / kit according to claim 4, characterized in that, It contains primers and / or probes for the specific detection of miR-145-5p.

6. A pharmaceutical composition for the prevention and / or treatment of pseudorabies virus infection, characterized in that, It contains an effective amount of miR-145-5p mimic and a pharmaceutically acceptable carrier.

7. The composition according to claim 6, characterized in that, Pharmaceutically acceptable carriers include liposomes, exosomes, or biodegradable nanoparticles.

8. A method for inhibiting pseudorabies virus replication in vitro, characterized in that, Introduce an effective amount of miR-145-5p mimic into cells infected or potentially infected with PRV.

Images