Application of NR4A1 inhibitor in preparation of drugs related to porcine reproductive and respiratory syndrome

Abstract

The invention discloses an application of an NR4A1 inhibitor in preparation of drugs related to porcine reproductive and respiratory syndrome, the invention proposes that the NR4A1 inhibitor DIM-C-pPhCO2Me can relieve intestinal injury caused by the porcine reproductive and respiratory syndrome for the first time, and the NR4A1 inhibitor can effectively improve abnormal death of intestinal cells caused by the porcine reproductive and respiratory syndrome. Experimental data show that the NR4A1 inhibitor DIM-C-pPhCO2Me can be used for effectively treating and reducing the porcine reproductive and respiratory syndrome and reducing the diarrhea rate of piglets; meanwhile, the porcine reproductive and respiratory syndrome prognosis can be effectively improved, and the growth performance of tolerant piglets is improved.

Description

Technical Field

[0001] The present invention relates to the field of veterinary medicine, and in particular to the application of NR4A1 inhibitor in the preparation of medicaments related to porcine reproductive and respiratory syndrome. Background Art

[0002] Porcine Reproductive and Respiratory Syndrome (PRRS) is a highly contagious disease caused by PRRS virus (PRRSV). The pathogen belongs to the family of Arteriviridae and is a single-stranded RNA virus that can be vertically transmitted through respiratory secretions, semen and placenta, destroying the host immune system function, leading to immunosuppression and increased risk of secondary infection.

[0003] However, its clinical manifestations are not limited to the respiratory tract and reproductive system. Intestinal symptoms and long-term damage to growth performance have attracted much attention in recent years. Intestinal symptoms such as diarrhea are particularly prominent in PRRS infection, especially in piglets and fattening pigs. About 30%-40% of cases are accompanied by watery diarrhea or mucous bloody stools. In severe cases, dehydration and electrolyte imbalance can lead to acute death. Studies have shown that PRRSV can infect intestinal macrophages and epithelial cells, destroy the intestinal barrier function, induce intestinal inflammatory response, and inhibit local immune response, leading to secondary infection of conditional pathogens (such as Escherichia coli and Salmonella), further aggravating the course of diarrhea.

[0004] Slow weight gain is the most significant long-term prognostic problem after PRRS infection: the surviving pigs in the acute infection still show persistent growth retardation after recovery. Studies have shown that even after the disappearance of viremia, it still takes weeks to months for growth performance to recover, and some pigs may permanently fall behind the normal growth curve.

[0005] Currently, supportive therapy is mostly used in clinical treatment. Although it can relieve gastrointestinal symptoms such as diarrhea, it cannot block the direct damage of the virus to the intestines, and there is a lack of specific drugs for gastrointestinal damage secondary to PRRSV. Summary of the invention

[0006] Purpose of the invention: The purpose of the present invention is to provide an application of an NR4A1 inhibitor DIM-C-pPhCO2Me in the preparation of a drug for treating or improving the prognosis of porcine reproductive and respiratory syndrome.

[0007] Technical solution: Use of the NR4A1 inhibitor of the present invention in the preparation of a drug for treating porcine reproductive and respiratory syndrome.

[0008] Preferably, the NR4A1 inhibitor is DIM-C-pPhCO2Me, whose molecular formula is C 25 H 20 N2O2.

[0009] Preferably, the drug contains an NR4A1 inhibitor as an active ingredient and a pharmaceutically acceptable excipient.

[0010] Preferably, the dosage form of the drug is a granule, tablet, capsule, oral liquid, pill, emulsion, suspension, injection, infusion or spray.

[0011] Preferably, the application is for the preparation of a drug for treating gastrointestinal symptoms of porcine reproductive and respiratory syndrome, and the gastrointestinal symptoms include any one or more of vomiting, diarrhea, nausea, and abdominal pain.

[0012] Use of the NR4A1 inhibitor according to the present invention in the preparation of a drug for improving the prognosis of porcine reproductive and respiratory syndrome.

[0013] Preferably, the NR4A1 inhibitor is DIM-C-pPhCO2Me, and its molecular formula is C 25 H 20 N2O2.

[0014] Preferably, the drug contains an NR4A1 inhibitor as an active ingredient and a pharmaceutically acceptable excipient.

[0015] Preferably, the dosage form of the drug is a granule, tablet, capsule, oral liquid, pill, emulsion, suspension, injection, infusion or spray.

[0016] Preferably, the application is for the preparation of a drug for improving the prognosis of surviving piglets with porcine reproductive and respiratory syndrome, and the improved prognosis includes at least one of average daily gain, feed-to-meat ratio, and body weight compliance rate.

[0017] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages: 1. DIM-C-pPhCO2Me can effectively treat and reduce porcine reproductive and respiratory syndrome and reduce the diarrhea rate of piglets; 2. DIM-C-pPhCO2Me can effectively improve the prognosis of porcine reproductive and respiratory syndrome and improve the growth performance of surviving piglets. Description of the Drawings

[0018] Figure 1 It is a fluorescence image of the survival of porcine ileal epithelial cells in different treatment groups after 36 hours of treatment;

[0019] Figure 2 It is a statistical result graph of the survival of porcine ileal epithelial cells in different treatment groups after 36 hours of treatment;

[0020] Figure 3 It is a statistical result graph of the relative cell viability of porcine ileal epithelial cells in different treatment groups after 48 hours of treatment;

[0021] Figure 4 Statistical result graph of DIM-C-pPhCO2Me improving the growth performance after porcine reproductive and respiratory syndrome

[0022] Figure 5 Statistical result graph of DIM-C-pPhCO2Me relieving diarrhea of porcine reproductive and respiratory syndrome Detailed implementation mode

[0023] The technical solution of the present invention will be further described below.

[0024] Example 1: The NR4A1 inhibitor DIM-C-pPhCO2Me relieves the damage of porcine reproductive and respiratory syndrome virus to intestinal cells

[0025] 1. Porcine ileal epithelial cells were inoculated in a 96-well cell culture plate at a density of 5×10 3 cells / well and incubated for 16 h to wait for cell attachment and further growth;

[0026] 2. A control group, a DIM-C-pPhCO2Me treatment group, a PRRSV treatment group, and a DIM-C-pPhCO2Me + PRRSV treatment group were set up. Among them, in the control group, the original medium was replaced with DMEM complete medium; in the DIM-C-pPhCO2Me treatment group, the original medium was replaced with DMEM complete medium containing DIM-C-pPhCO2Me, and the final concentrations of DIM-C-pPhCO2Me were 1, 5, and 10 μM respectively; in the PRRSV treatment group, the original medium was replaced with DMEM complete medium containing PRRSV, and the final concentration of PRRSV was 10 4 TCID 50 / mL (porcine ileal epithelial cells); in the DIM-C-pPhCO2Me + PRRSV treatment group, the original medium was replaced with DMEM complete medium containing both DIM-C-pPhCO2Me and PRRSV, the final concentrations of DIM-C-pPhCO2Me were 1, 5, and 10 μM respectively, and the final concentration of PRRSV was 10 4 TCID 50 / mL (porcine ileal epithelial cells).

[0027] 3. After 36 h of treatment, calcein-AM was used to stain live cells, and a fluorescence microscope was used for observation; after 48 h of treatment, it was replaced with 90 μL of fresh DMEM complete medium, and 10 μL of CCK8 was added to each well, and incubated in the cell culture incubator in the dark for 15 min. The absorbance was measured at 450 nm, and the relative cell viability was calculated and normalized through formula (1):

[0028] Relative cell viability = (A s -A b) / (Ac - A b )×100% (1).

[0029] A s 、A b and A c are the absorbance values of the sample, blank control, and negative control, respectively.

[0030] The results of fluorescence microscopy observation are as shown in Figure 1 , and the statistical results are as shown in Figure 2 . DIM-C-pPhCO2Me at a concentration of 10 μM does not cause damage to porcine ileal epithelial cells. At the same time, DIM-C-pPhCO2Me at concentrations of 1 and 5 μM can effectively alleviate the damage of PRRSV to intestinal cells. The detection of relative cell viability verified this point, and the results are as shown in Figure 3 . DIM-C-pPhCO2Me treatment at a concentration of 5 μM has no obvious cytotoxicity but can effectively alleviate cell death caused by PRRSV.

[0031] Example 2 DIM-C-pPhCO2Me Alleviates Porcine Reproductive and Respiratory Syndrome Symptoms and Improves Prognosis

[0032] 1. Twenty experimental piglets were purchased from Zhou Chunlong Farm in Pukou District, Nanjing City and raised in Lihong Livestock and Poultry Breeding Center in Jiangbei New Area, Nanjing City. During the experiment, the indoor temperature was maintained at 24 ± 2 °C. All piglets had free access to food and water. All piglets were randomly assigned to a control group, a DIM-C-pPhCO2Me treatment group, a PRRSV treatment group, and a DIM-C-pPhCO2Me + PRRSV treatment group, with 5 piglets in each group.

[0033] 2. The piglets in the PRRSV treatment group and the DIM-C-pPhCO2Me + PRRSV treatment group were treated by intranasal drip with 2 mL of PRRSV at a concentration of 10 4 TCID 50 / mL (porcine ileal epithelial cells). The control group and the DIM-C-pPhCO2Me treatment group were treated by intranasal drip with 2 mL of normal saline. At the same time, the DIM-C-pPhCO2Me treatment group and the DIM-C-pPhCO2Me + PRRSV treatment group were intravenously injected with a dose of 1 mg / kg of DIM-C-pPhCO2Me solution daily; the control group of PRRSV and the treatment group were intravenously injected with an equal amount of normal saline daily.

[0034] 3. DIM-C-pPhCO2Me and normal saline were continuously injected for 14 days, and the diarrhea situation and body weight were recorded daily.

[0035] The recorded results of the piglet body weight are as shown in Figure 4As shown, the first day is the first day of drug administration after the successful establishment of the PRRS model, manifested as obvious listlessness. The change in the body weight of piglets reflects that DIM-C-pPhCO2Me can effectively relieve the weight loss caused by PRRSV and can effectively improve the growth performance of piglets.

[0036] The diarrhea situation of piglets is as Figure 5 shown. DIM-C-pPhCO2Me can effectively inhibit the intestinal damage of piglets caused by PRRSV and relieve the diarrhea situation of piglets.

Claims

1. Use of an NR4A1 inhibitor in the preparation of a therapeutic drug for porcine reproductive and respiratory syndrome.

2. The application according to claim 1, characterized in that, The NR4A1 inhibitor is DIM-C-pPhCO2Me, and its molecular formula is C 25 H 20 N2O2.

3. The application according to claim 1, wherein The drug contains an NR4A1 inhibitor as an active ingredient and pharmaceutically acceptable excipients.

4. The application according to claim 1, wherein The dosage form of the drug is granule, tablet, capsule, oral liquid, pill, emulsion, suspension, injection, infusion or spray.

5. The application according to any one of claims 1-4, characterized in that The application is the use in the preparation of a drug for treating gastrointestinal symptoms of porcine reproductive and respiratory syndrome, and the gastrointestinal symptoms include any one or more of vomiting, diarrhea, nausea, and abdominal pain.

6. Use of an NR4A1 inhibitor in the preparation of a drug for improving the prognosis of porcine reproductive and respiratory syndrome.

7. The application according to claim 6, characterized in that The NR4A1 inhibitor is DIM-C-pPhCO2Me, and its molecular formula is C 25 H 20 N2O2.

8. The application according to claim 6, wherein, The drug contains an NR4A1 inhibitor as an active ingredient and pharmaceutically acceptable excipients.

9. The application according to claim 6, wherein The dosage form of the drug is granule, tablet, capsule, oral liquid, pill, emulsion, suspension, injection, infusion or spray.

10. The application according to any one of claims 6-9, characterized in that, The application is the use in the preparation of a drug for improving the prognosis of surviving piglets with porcine reproductive and respiratory syndrome, and the improved prognosis includes at least one of average daily weight gain, feed-to-meat ratio, and body weight compliance rate.

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