A traditional Chinese medicine composition for preventing and / or treating porcine reproductive and respiratory syndrome, a preparation method thereof, and a traditional Chinese medicine preparation

Abstract

This invention relates to the field of veterinary preventive and therapeutic drugs. The invention provides a traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome (PRRS) and its preparation method, comprising the following components: Indigo naturalis, Forsythia suspensa, Arctium lappa, Dryopteris crassirhizoma, Lithospermum erythrorhizon, Taraxacum mongolicum, Rheum palmatum, Scutellaria barbata, Glycyrrhiza uralensis, Hordeum vulgare, Bupleurum chinense, and Terminalia chebula. The invention also provides a traditional Chinese medicine preparation comprising the aforementioned composition. Using the above-mentioned components in combination with the technical solution of this invention can effectively enhance the inhibitory effect on pathogenic microorganisms and the anti-PRRSV virus effect, improve immunity, and enhance the disease resistance of pigs; alleviate symptoms such as depression and loss of appetite caused by PRRSV, and regulate the gastrointestinal health of animals; at the same time, it has few toxic side effects and is less likely to induce drug resistance.

Description

Technical Field

[0001] This invention relates to the field of veterinary preventive and therapeutic drugs, and in particular to a traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, its preparation method, and its traditional Chinese medicine preparation. Background Technology

[0002] Porcine reproductive and respiratory syndrome (PRRS), also known as "blue ear disease," is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV). It is characterized by fever and anorexia in infected pigs, late-term abortion, premature birth, stillbirth, weak piglets, and mummified fetuses in pregnant sows, and respiratory disorders in pigs of all ages (especially piglets). This disease is also immunosuppressive, weakening the immune system of infected pigs and making them highly susceptible to secondary and concurrent viral, bacterial, and parasitic diseases. This can lead to mixed or secondary infections of multiple diseases, causing mass mortality and significant economic losses to the pig farming industry.

[0003] Currently, the common practice for preventing and controlling porcine reproductive and respiratory syndrome (PRRS) involves vaccinating pigs against the PRRS virus and treating them with antibiotics. However, the results are not ideal, and the use of synthetic antiviral drugs can produce certain toxic side effects, impacting normal cells in the body. Therefore, it is essential to develop a drug that has better preventive and therapeutic effects against PRRS with fewer toxic side effects. Summary of the Invention

[0004] The purpose of this invention is to provide a traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome (PRRS). By using the components of this invention in combination, it can effectively enhance the inhibitory effect on pathogenic microorganisms and the anti-PRRSV virus effect, improve the immunity of pigs, and enhance the disease resistance of pigs; alleviate symptoms such as depression and loss of appetite caused by PRRSV, and regulate the gastrointestinal health of animals; at the same time, it has few toxic side effects and is not prone to drug resistance.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following components in parts by weight: 22-35 parts Indigo Naturalis, 20-26 parts Forsythia suspensa, 13-18 parts Arctium lappa, 18-30 parts Dryopteris crassirhizoma, 16-22 parts Lithospermum erythrorhizon, 10-17 parts Taraxacum mongolicum, 10-15 parts Rheum palmatum, 11-16 parts Scutellaria barbata, 9-14 parts Glycyrrhiza uralensis, 9-13 parts Hordeum vulgare, 10-13 parts Bupleurum chinense, and 7-10 parts Terminalia chebula.

[0007] As a preferred embodiment, the composition includes the following components in parts by weight: 24-32 parts Indigo Naturalis, 21-25 parts Forsythia suspensa, 14-17 parts Arctium lappa, 19-28 parts Dryopteris crassirhizoma, 17-20 parts Lithospermum erythrorhizon, 12-16 parts Taraxacum mongolicum, 11-14 parts Rheum palmatum, 12-15 parts Scutellaria barbata, 10-13 parts Glycyrrhiza uralensis, 10-12 parts Hordeum vulgare, 10.5-12 parts Bupleurum chinense, and 8-9.5 parts Terminalia chebula.

[0008] This invention provides a method for preparing the traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following steps:

[0009] (1) Take rhubarb, forsythia, dandelion and licorice, cut them into sections and mix them together. Add water and decoct. Filter to obtain filtrate A.

[0010] (2) Take burdock seed, red-striped gromwell root, dryopteris crassirhizoma, scutellaria barbata, malt, bupleurum root and green fruit, crush them separately and mix them, reflux extract, filter to obtain filtrate B;

[0011] (3) Mix the obtained filtrate A, filtrate B and indigo and concentrate to obtain the final product.

[0012] Preferably, the amount of water added in step (1) is 3-7 times the total mass of Astragalus membranaceus, Forsythia suspensa, Taraxacum mongolicum and Glycyrrhiza uralensis; the decoction time is 50-80 min / time, and the number of decoctions is 1-3 times.

[0013] Preferably, the solvent used for reflux extraction in step (2) is an ethanol solution with a volume concentration of 65%-85%, and the volume mass ratio of the solvent to the total mass of burdock seed, red-striped gromwell root, Dryopteris crassirhizoma, Scutellaria barbata, malt, Bupleurum chinense and green fruit is (8-15) mL:(1-3) g.

[0014] Preferably, the reflux extraction time in step (2) is 60-100 min / time, and the number of reflux extractions is 1-4 times.

[0015] The present invention also provides a traditional Chinese medicine preparation for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the traditional Chinese medicine composition prepared by the above-described method.

[0016] Preferably, the dosage form of the traditional Chinese medicine preparation includes oil, powder, granules or solution.

[0017] By adopting the above technical solution, the present invention has the following beneficial effects:

[0018] In this invention, the combination of Indigo Naturalis with Arctium lappa, Forsythia suspensa, Dryopteris crassirhizoma, and Terminalia chebula in the technical solution of the present invention synergistically enhances the effects of clearing heat and detoxifying, antibacterial, and antiviral. Licorice root, in addition to containing various effective detoxifying components, can also harmonize the properties of the overall formula and is used as an adjuvant. Through the combined use of the components of this invention, the inhibitory effects on pathogenic microorganisms and the anti-PRRSV virus can be enhanced. It can also alleviate symptoms such as depression and loss of appetite, regulate the gastrointestinal health of animals, improve immunity, and enhance the disease resistance of pigs, thereby effectively preventing and treating porcine reproductive and respiratory syndrome (PRRS). The traditional Chinese medicine composition of this invention not only has good preventive and therapeutic effects but also has few toxic side effects and is less likely to induce drug resistance. Detailed Implementation

[0019] This invention provides a traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following components: indigo naturalis, forsythia suspensa, burdock seed, Dryopteris crassirhizoma, Lithospermum erythrorhizon, dandelion, rhubarb, Scutellaria barbata, licorice, malt, Bupleurum chinense, and Terminalia chebula.

[0020] In this invention, the preferred weight percentage of the indigo naturalis is 22-35 parts, more preferably 24-32 parts, and even more preferably 30 parts; the preferred weight percentage of the forsythia suspensa is 20-26 parts, more preferably 21-25 parts, and even more preferably 23 parts; the preferred weight percentage of the burdock seed is 13-18 parts, more preferably 14-17 parts, and even more preferably 15.5 parts; the preferred weight percentage of the Dryopteris crassirhizoma is 18-30 parts, more preferably 19-28 parts, and even more preferably 25 parts; the preferred weight percentage of the Lithospermum erythrorhizon is 16-22 parts, more preferably 17-20 parts, and even more preferably 18 parts; and the preferred weight percentage of the dandelion is 10-17 parts, more preferably 12-16 parts, and even more preferably... 14 parts; the weight of rhubarb is preferably 10-15 parts, more preferably 11-14 parts, and even more preferably 13 parts; the weight of Scutellaria barbata is preferably 11-16 parts, more preferably 12-15 parts, and even more preferably 13.5 parts; the weight of licorice is preferably 9-14 parts, more preferably 10-13 parts, and even more preferably 12 parts; the weight of malt is preferably 9-13 parts, more preferably 10-12 parts, and even more preferably 10.5 parts; the weight of Bupleurum chinense is preferably 10-13 parts, more preferably 10.5-12 parts, and even more preferably 11.5 parts; the weight of Terminalia chebula is preferably 7-10 parts, more preferably 8-9.5 parts, and even more preferably 8.5 parts.

[0021] In this invention, the indigo naturalis is a dried powder, lumps, or granules obtained by processing the leaves or stems of *Strobilanthes cusia* (Acanthaceae), *Polygonum tinctorium* (Polygonaceae), and *Isatis tinctoria* (Brassicaceae). It has a salty and cold nature and possesses the effects of clearing heat and detoxifying, and resisting pathogenic microorganisms.

[0022] In this invention, the preparation method of indigo is as follows: (1) Take the leaves or stems of *Strobilanthes cusia* (Acanthaceae), *Polygonum tinctorium* (Polygonaceae), or *Isatis tinctoria* (Brassicaceae), and extract the corresponding fresh leaves in hot water. Filter to remove leaf residue and obtain filtrate; (2) Add an acid solution to the filtrate for hydrolysis. After hydrolysis, a hydrolyzed mixture is obtained; (3) Add an alkaline solution to adjust the pH of the hydrolyzed mixture to 6-8, let it stand, discard the supernatant, take the precipitate, wash until neutral, and dry to obtain indigo. In this invention, the acid solution is preferably acetic acid or hydrochloric acid, more preferably hydrochloric acid; the alkaline solution is preferably sodium hydroxide or ammonia, more preferably sodium hydroxide. The preferred temperature of the hot water in this invention is 60-90℃, more preferably 70-85℃, and even more preferably 80℃; the preferred volume-to-mass ratio of the hot water to the fresh leaves is (10-20) mL:(0.5-1.5) g, more preferably (12-18) mL:(0.8-1.3) g, and even more preferably 14 mL:1.2 g; the preferred extraction time is 20-30 min / time, more preferably 22-28 min / time, and even more preferably 25 min / time; the preferred number of extractions is 2-4 times, and even more preferably 3 times. The moisture content of the indigo in this invention does not exceed 7%.

[0023] In this invention, the burdock seed is pungent, bitter, and cold in nature; it enters the lung and stomach meridians; the medicinal part is the dried, mature fruit of the biennial herbaceous plant *Arctium lappa* of the Asteraceae family. It has antipyretic, sedative, analgesic, antimicrobial, and immunomodulatory effects. In this invention, the forsythia is slightly cold in nature and bitter in taste; it enters the lung, heart, and small intestine meridians; it has antimicrobial, antipyretic, and anti-inflammatory effects. In this invention, the Dryopteris crassirhizoma, also known as Dryopteris crassirhizoma, is the rhizome and petiole remnants of the Dryopteris crassirhizoma plant of the Dryopteris family; it is bitter and slightly cold in nature; it enters the liver and stomach meridians; it has antimicrobial, anthelmintic, and anticancer effects. In this invention, the green fruit is a traditional Chinese medicine, bitter, pungent, and cold in nature; it enters the heart, lung, and large intestine meridians; it contains flavonoids, polyphenols, gallic acid, volatile oils, and other components, and has heat-clearing, detoxifying, antiviral, and antibacterial effects. In this invention, indigo naturalis is used in combination with burdock fruit, forsythia, Dryopteris crassirhizoma, and Terminalia chebula, which significantly enhances the effects of resisting pathogenic microorganisms, detoxification, and antipyresis.

[0024] In this invention, the red-striped gromwell, also known as gromwell, is a plant belonging to the genus Gromwell in the family Boraginaceae. It is cold in nature and has a sweet and salty taste; it contains chemical components such as acetylshikonin, β-hydroxyisovalerylshikonin, shikonin, and β,β'-dimethylacryloylshikonin, and has the effects of clearing heat and detoxifying.

[0025] In this invention, the dandelion powder contains abundant vitamin B2, taraxasterol, caffeic acid, etc., and dandelion can also promote phagocytosis by phagocytes, enhance the function of immune cells and humoral immunity, thereby improving human immunity. When combined with forsythia and other herbs, it can better enhance human resistance and immunity, while also strengthening antibacterial and anti-inflammatory effects.

[0026] In this invention, Bupleurum can soothe the liver and relieve depression, alleviating symptoms such as emotional instability, abdominal distension and belching, and loss of appetite.

[0027] In this invention, the Scutellaria barbata is cold in nature and sour in taste. The whole herb contains a variety of vitamins, trace elements, amino acids, flavonoids and other components, and has antimicrobial, antipyretic and anti-inflammatory effects.

[0028] In this invention, the malt is sweet and neutral, and can promote digestion, strengthen the spleen and stomach, and is especially good at promoting the digestion of starchy foods; the α and β amylase it contains can break down starch into maltose and dextrin; the B vitamins contained in malt also promote the secretion of gastric acid and pepsin, which can help digestion.

[0029] The present invention also provides a method for preparing the traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following steps:

[0030] (1) Take rhubarb, forsythia, dandelion and licorice, cut them into sections and mix them together. Add water and decoct. Filter to obtain filtrate A.

[0031] (2) Take burdock seed, red-striped gromwell root, dryopteris crassirhizoma, scutellaria barbata, malt, bupleurum root and green fruit, crush them separately and mix them, reflux extract, filter to obtain filtrate B;

[0032] (3) Mix the obtained filtrate A, filtrate B and indigo and concentrate to obtain the final product.

[0033] In this invention, fresh, intact, insect-free, mold-free, and unmixed raw material components are selected. The selected raw materials are washed with clean water and then dried (excluding indigo). Preferably, the raw materials are dried to a moisture content of less than 13% to better preserve the content of effective components in the raw materials, reduce the loss of aromatic and medicinal components, and prevent mold growth.

[0034] In this invention, dried rhubarb, forsythia, dandelion, and licorice are cut into segments, preferably 0.5-1.5 cm in length, more preferably 0.8-1.3 cm, and even more preferably 1 cm. The cut ingredients are then mixed and decocted with water. The amount of water added is preferably 3-7 times the total mass of the astragalus, forsythia, dandelion, and licorice, more preferably 3.5-6 times, and even more preferably 5 times. The decoction time is preferably 50-80 min / time, more preferably 55-70 min / time, and even more preferably 60 min / time. The number of decoctions is preferably 1-3 times, more preferably 2 times. After each decoction, the mixture is filtered to separate the dregs, and the filtrate is collected. The filtrates are then combined to obtain filtrate A.

[0035] In this invention, dried burdock seeds, Lithospermum erythrorhizon, Dryopteris crassirhizoma, Scutellaria barbata, malt, Bupleurum chinense, and Terminalia chebula are pulverized separately, then sieved, and the sieve residue is collected and mixed. The mesh size of the sieve used in this invention is preferably 50-100 mesh, more preferably 60-80 mesh, and even more preferably 70 mesh. In this invention, pulverized burdock seeds, Lithospermum erythrorhizon, Dryopteris crassirhizoma, Scutellaria barbata, malt, Bupleurum chinense, and Terminalia chebula are mixed and then subjected to reflux extraction. The solvent used for reflux extraction is preferably an ethanol solution, with a volume concentration preferably of 65%-85%, more preferably 70%-80%, and even more preferably 75%. The volume-to-mass ratio of the solvent to the total mass of burdock seeds, Lithospermum erythrorhizon, Dryopteris crassirhizoma, Scutellaria barbata, malt, Bupleurum chinense, and Terminalia chebula is preferably (8-15) mL:(1-3) g, more preferably (10-18) mL:(1.4-2.7) g, and even more preferably 14 mL:1.5 g. The reflux extraction time is preferably 60-100 min / time, more preferably 70-90 min / time, and even more preferably 75 min / time. The number of reflux extractions is preferably 1-4 times, more preferably 2-3 times.

[0036] In this invention, the obtained filtrate A, filtrate B and indigo are mixed evenly and then concentrated, preferably to a relative density of 1.05-1.2, and more preferably 1.10-1.15.

[0037] The present invention also provides a traditional Chinese medicine preparation for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the traditional Chinese medicine composition prepared by the above-described method.

[0038] In this invention, the dosage forms of the traditional Chinese medicine preparations include oils, powders, granules, or solutions. In this invention, the traditional Chinese medicine composition can be prepared in oil, powder, granule, or solution dosage forms by adding conventional excipients using conventional methods.

[0039] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0040] Example 1

[0041] A traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following components: 30 parts Indigo Naturalis, 23 parts Forsythia suspensa, 15.5 parts Arctium lappa, 25 parts Dryopteris crassirhizoma, 18 parts Lithospermum erythrorhizon, 14 parts Taraxacum mongolicum, 13 parts Rheum palmatum, 13.5 parts Scutellaria barbata, 12 parts Glycyrrhiza uralensis, 10.5 parts Hordeum vulgare, 11.5 parts Bupleurum chinense, and 8.5 parts Terminalia chebula;

[0042] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps:

[0043] (1) Take the stems and leaves of the plant Strobilanthes cusia in the Acanthaceae family and place them in hot water at 80℃ at a volume-to-mass ratio of 14mL:1.2g. Extract three times for 25min each time. Filter to remove the leaf residue and obtain the filtrate. Add 3% hydrochloric acid solution to the obtained filtrate and hydrolyze for 50min to obtain a hydrolyzed mixture. Add 3% alkaline solution to the obtained hydrolyzed mixture to adjust the pH to 7. Let it stand and discard the supernatant. Take the precipitate, wash it with water until neutral, and dry it to obtain indigo naturalis with a moisture content of no more than 7%.

[0044] (2) Select fresh, whole rhubarb, forsythia, dandelion and licorice that are free from insects, mold and impurities. Wash them with clean water and dry them until the moisture content is less than 13%. Then cut them into 1cm lengths and mix them together. Add water at 5 times the total mass of rhubarb, forsythia, dandelion and licorice and decoct for 60min / time, twice. Filter after each decoction, separate the dregs, collect the filtrate, and then combine the filtrates to obtain filtrate A.

[0045] (3) Select fresh, intact, insect-free, mold-free, and unmixed burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Wash them with clean water and dry them until the moisture content is less than 13%. Crush them separately and pass them through a 70-mesh sieve. Collect the sieve material and mix it. Then add 75% ethanol solution at a volume mass ratio of 14 mL to 1.5 g of the total mass of burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Reflux extraction is performed for 75 min / time, and the extraction is performed twice.

[0046] (4) Mix the obtained filtrate A, filtrate B and indigo evenly, and concentrate to a relative density of 1.10 to obtain the final product.

[0047] Example 2

[0048] A traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following components: 22 parts indigo naturalis, 22 parts forsythia suspensa, 13 parts burdock fruit, 18 parts Dryopteris crassirhizoma, 16 parts Lithospermum erythrorhizon, 10 parts dandelion, 15 parts rhubarb, 11 parts Scutellaria barbata, 9 parts licorice, 9 parts malt, 10 parts Bupleurum chinense, and 7 parts Terminalia chebula.

[0049] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps:

[0050] (1) Take the stems and leaves of the plant Strobilanthes cusia in the Acanthaceae family and place them in hot water at 80℃ at a volume-to-mass ratio of 14mL:1.2g. Extract three times for 25min each time. Filter to remove the leaf residue and obtain the filtrate. Add 3% hydrochloric acid solution to the obtained filtrate and hydrolyze for 50min to obtain a hydrolyzed mixture. Add 3% alkaline solution to the obtained hydrolyzed mixture to adjust the pH to 7. Let it stand and discard the supernatant. Take the precipitate, wash it with water until neutral, and dry it to obtain indigo naturalis with a moisture content of no more than 7%.

[0051] (2) Select fresh, whole rhubarb, forsythia, dandelion and licorice that are free from insects, mold and impurities. Wash them with clean water and dry them until the moisture content is less than 13%. Then cut them into 1cm lengths and mix them together. Add water at 5 times the total mass of rhubarb, forsythia, dandelion and licorice and decoct for 60min / time, twice. Filter after each decoction, separate the dregs, collect the filtrate, and then combine the filtrates to obtain filtrate A.

[0052] (3) Select fresh, intact, insect-free, mold-free, and unmixed burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Wash them with clean water and dry them until the moisture content is less than 13%. Crush them separately and pass them through a 70-mesh sieve. Collect the sieve material and mix it. Then add 75% ethanol solution at a volume mass ratio of 14 mL to 1.5 g of the total mass of burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Reflux extraction is performed for 75 min / time, and the extraction is performed twice.

[0053] (4) Mix the obtained filtrate A, filtrate B and indigo evenly, and concentrate to a relative density of 1.10 to obtain the final product.

[0054] Example 3

[0055] A traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following components: 35 parts indigo naturalis, 26 parts forsythia suspensa, 18 parts burdock fruit, 30 parts Dryopteris crassirhizoma, 22 parts Lithospermum erythrorhizon, 17 parts dandelion, 15 parts rhubarb, 16 parts Scutellaria barbata, 14 parts licorice root, 13 parts malt, 13 parts Bupleurum chinense, and 10 parts Terminalia chebula.

[0056] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps:

[0057] (1) Take the stems and leaves of the plant Strobilanthes cusia in the Acanthaceae family and place them in hot water at 80℃ at a volume-to-mass ratio of 14mL:1.2g. Extract three times for 25min each time. Filter to remove the leaf residue and obtain the filtrate. Add 3% hydrochloric acid solution to the obtained filtrate and hydrolyze for 50min to obtain a hydrolyzed mixture. Add 3% alkaline solution to the obtained hydrolyzed mixture to adjust the pH to 7. Let it stand and discard the supernatant. Take the precipitate, wash it with water until neutral, and dry it to obtain indigo naturalis with a moisture content of no more than 7%.

[0058] (2) Select fresh, whole rhubarb, forsythia, dandelion and licorice that are free from insects, mold and impurities. Wash them with clean water and dry them until the moisture content is less than 13%. Then cut them into 1cm lengths and mix them together. Add water at 5 times the total mass of rhubarb, forsythia, dandelion and licorice and decoct for 60min / time, twice. Filter after each decoction, separate the dregs, collect the filtrate, and then combine the filtrates to obtain filtrate A.

[0059] (3) Select fresh, intact, insect-free, mold-free, and unmixed burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Wash them with clean water and dry them until the moisture content is less than 13%. Crush them separately and pass them through a 70-mesh sieve. Collect the sieve material and mix it. Then add 75% ethanol solution at a volume mass ratio of 14 mL to 1.5 g of the total mass of burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Reflux extraction is performed for 75 min / time, and the extraction is performed twice.

[0060] (4) Mix the obtained filtrate A, filtrate B and indigo evenly, and concentrate to a relative density of 1.10 to obtain the final product.

[0061] Example 4

[0062] A traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following components: 30 parts Indigo Naturalis, 23 parts Forsythia suspensa, 15.5 parts Arctium lappa, 25 parts Dryopteris crassirhizoma, 18 parts Lithospermum erythrorhizon, 14 parts Taraxacum mongolicum, 13 parts Rheum palmatum, 13.5 parts Scutellaria barbata, 12 parts Glycyrrhiza uralensis, 10.5 parts Hordeum vulgare, 11.5 parts Bupleurum chinense, and 8.5 parts Terminalia chebula;

[0063] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps:

[0064] (1) Take the stems and leaves of Polygonum indicum (Polygonaceae) at a volume-to-mass ratio of 14 mL: 1.2 g and place them in hot water at 80℃. Extract three times for 25 min each time. Filter to remove the leaf residue and obtain the filtrate. Add 3% hydrochloric acid solution to the obtained filtrate and hydrolyze for 50 min to obtain the hydrolyzed mixture. Add 3% alkaline solution to the obtained hydrolyzed mixture to adjust the pH to 7. Let it stand and discard the supernatant. Take the precipitate, wash it with water until neutral, and dry it to obtain indigo naturalis with a moisture content of no more than 7%.

[0065] (2) Select fresh, whole rhubarb, forsythia, dandelion and licorice that are free from insects, mold and impurities. Wash them with clean water and dry them until the moisture content is less than 13%. Then cut them into 1cm lengths and mix them together. Add water at 3 times the total mass of rhubarb, forsythia, dandelion and licorice and decoct for 80min / time. Decoction is performed once. After each decoction, filter to separate the dregs and collect the filtrate. Then combine the filtrates to obtain filtrate A.

[0066] (3) Select fresh, intact, insect-free, mold-free, and unmixed burdock seeds, red-striped gromwell root, drynaria fortunei, scutellaria barbata, malt, bupleurum chinense, and green fruit. Wash them with clean water and dry them until the moisture content is less than 13%. Crush them separately and pass them through a 70-mesh sieve. Collect the sieve material and mix it. Then add 65% ethanol solution at a volume mass ratio of 8 mL:3 g to the total mass of burdock seeds, red-striped gromwell root, drynaria fortunei, scutellaria barbata, malt, bupleurum chinense, and green fruit. Reflux extraction is performed for 60 min / time, for 4 times.

[0067] (4) Mix the obtained filtrate A, filtrate B and indigo evenly, and concentrate to a relative density of 1.10 to obtain the final product.

[0068] Example 5

[0069] A traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, comprising the following components: 30 parts Indigo Naturalis, 23 parts Forsythia suspensa, 15.5 parts Arctium lappa, 25 parts Dryopteris crassirhizoma, 18 parts Lithospermum erythrorhizon, 14 parts Taraxacum mongolicum, 13 parts Rheum palmatum, 13.5 parts Scutellaria barbata, 12 parts Glycyrrhiza uralensis, 10.5 parts Hordeum vulgare, 11.5 parts Bupleurum chinense, and 8.5 parts Terminalia chebula;

[0070] The preparation method of the above-mentioned traditional Chinese medicine composition includes the following steps:

[0071] (1) Take the stems and leaves of Isatis indigotica, a plant of the Brassicaceae family, and place them in hot water at 80℃ at a volume-to-mass ratio of 14mL:1.2g. Extract three times for 25min each time. Filter to remove the leaf residue and obtain the filtrate. Add 3% hydrochloric acid solution to the obtained filtrate and hydrolyze for 50min to obtain a hydrolyzed mixture. Add 3% alkaline solution to the obtained hydrolyzed mixture to adjust the pH to 7. Let it stand and discard the supernatant. Take the precipitate, wash it with water until neutral, and dry it to obtain indigo naturalis with a moisture content of no more than 7%.

[0072] (2) Select fresh, whole rhubarb, forsythia, dandelion and licorice that are free from insects, mold and impurities. Wash them with clean water and dry them until the moisture content is less than 13%. Then cut them into 1cm lengths and mix them together. Add water at 7 times the total mass of rhubarb, forsythia, dandelion and licorice and decoct for 50min / time, twice. Filter after each decoction, separate the dregs, collect the filtrate, and then combine the filtrates to obtain filtrate A.

[0073] (3) Select fresh, intact, insect-free, mold-free, and unmixed burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Wash them with clean water and dry them until the moisture content is less than 13%. Crush them separately and pass them through a 70-mesh sieve. Collect the sieve material and mix it. Then add 65% ethanol solution at a volume mass ratio of 20 mL: 1 g to the total mass of burdock seeds, red-striped gromwell root, drynaria rhizome, scutellaria barbata, malt, bupleurum root, and green fruit. Reflux extraction is performed for 100 min / time, and the extraction is performed twice.

[0074] (4) Mix the obtained filtrate A, filtrate B and indigo evenly, and concentrate to a relative density of 1.15 to obtain the final product.

[0075] Comparative Example 1

[0076] Similar to Example 1, only "30 parts Indigo Naturalis" and "25 parts Dryopteris crassirhizoma" were removed.

[0077] Comparative Example 2

[0078] Similar to Example 1, only "10.5 parts malt and 11.5 parts Bupleurum" were removed.

[0079] Comparative Example 3

[0080] Similar to Example 1, except that "then add 75% ethanol solution at a volume-to-mass ratio of 14 mL:1.5 g to the total mass of burdock seed, red-striped gromwell root, drynaria fortunei, scutellaria barbata, malt, bupleurum, and green plum, and reflux extract for 75 min / time, twice" is replaced with "then add 75% ethanol solution at a volume-to-mass ratio of 30 mL:1 g to the total mass of burdock seed, red-striped gromwell root, drynaria fortunei, scutellaria barbata, malt, bupleurum, and green plum, and reflux extract for 30 min / time, twice".

[0081] Comparative Example 4

[0082] Similar to Example 1, except that "then add 75% ethanol solution at a volume-to-mass ratio of 14 mL to 1.5 g of the total mass of burdock seed, red-striped gromwell root, dryopteris crassirhizoma, scutellaria barbata, malt, bupleurum chinense, and green plum, and reflux extract for 75 min / time, twice" is replaced with "then add water and decoct at a volume-to-mass ratio of 14 mL to 1.5 g of the total mass of burdock seed, red-striped gromwell root, dryopteris crassirhizoma, scutellaria barbata, malt, bupleurum chinense, and green plum, for 75 min / time, twice".

[0083] Experimental Example 1

[0084] (I) Cytotoxicity test

[0085] (1) Marc-145 cells (purchased from the Kunming Cell Bank of the Chinese Academy of Sciences) were digested and passaged using standard methods, and the cell number was adjusted to 1×10⁻⁶. 6 After the cells were seeded at a density of 100 μL / mL, they were inoculated into 96-well cell culture plates and cultured in an incubator at 37°C and 5% CO2 until a monolayer was formed.

[0086] (2) The traditional Chinese medicine preparation obtained in Example 1 was serially diluted 2-fold starting from 1000 μg / mL to eight concentration gradients (2 0 2 -1 2 -2 2 -3 2 -4 2 -5 2 -6 2 -7 ), add 0.1 mL per well, for a total of 4 replicates, into the 96-well plate of step (1), and set up a control group with only 100 μL of 2% cell maintenance medium. Incubate in an incubator at 37°C and 5% CO2.

[0087] (3) After culturing for 72 hours, discard the supernatant, add 20 μL of MTT to each well, and incubate in an incubator at 37°C and 5% CO2 for 4 hours.

[0088] (4) Discard the MTT, add 150 μL of DMSO to each well, and incubate in an incubator at 37℃ and 5% CO2 for 5-10 min.

[0089] (5) The OD value at 570 nm was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the cell viability was calculated. See Table 1.

[0090] Cell viability rate = OD of experimental group 570nm Value / Cell control group OD 570nm Value × 100%.

[0091] Table 1. Cell viability rates of different concentrations of traditional Chinese medicine preparations.

[0092]

[0093] Table 1 shows that high concentrations of the aforementioned traditional Chinese medicine preparations have certain toxicity to Marc-145 cells; however, the degree of cell damage gradually decreases as the concentration decreases. (OD2000 was compared with the cell control group.) 570nm The highest concentration with no significant difference was taken as the maximum safe cellular concentration of the traditional Chinese medicine preparation, which is 125 μg / mL. SPSS analysis of the data showed that the half-maximal toxicity concentration (TC) was... 50 It was 1028.23 μg / mL. (two)

[0095] The PRRSVYN-1 virus strain was provided by the Animal Infectious Disease Laboratory of Yunnan Agricultural University. The virulence of the virus on Marc-145 cells was 10⁻⁵.⁵ TCID⁻¹. 50 / 0.1mL.

[0096] 100TCID 50 PRRSV was seeded onto a monolayer of Marc-145 cells at 100 μL / well, incubated at 37°C for 2 h, the virus solution was discarded, and the cells were washed three times with PBS. Starting at the maximum safe concentration, five serially diluted concentrations (125 μg / mL, 62.5 μg / mL, 31.25 μg / mL, 15.63 μg / mL, and 7.81 μg / mL) of the traditional Chinese medicine preparations prepared in Example 1 and Comparative Examples 1-4 were added at 100 μL / well, with each dilution replicated in four wells. All wells were then incubated at 37°C in a 5% CO2 incubator, and the time and extent of cytopathic effects were observed daily. The half-maximal inhibitory concentration (IC50) for different groups was calculated using the Reed-Muench method based on the cytopathic effect. 50 And calculate the index SI. See Table 2 for details.

[0097] Table 2. Half-maximal inhibitory concentrations (IC50) for different groups 50 and SI

[0098] Group <![CDATA[IC 50 (μg / mL)]]> SI Example 1 Group 78.49 13.1 Comparative Example 1 316.22 6.5 Comparative Example 2 226.63 7.9 Comparative Example 3 Groups 157.50 9.8 Comparative Example 4 Groups 107.16 11.2

[0099] As shown in Table 2, different groups of traditional Chinese medicine preparations were able to inhibit the proliferation of PRRSV in Marc-145 cells within a certain concentration range. Among them, the half-maximal inhibitory concentration (IC50) of the group in Example 1 was the highest. 50 The SI index is relatively the largest, indicating that the drug safety range of the traditional Chinese medicine preparations in Example 1 is relatively the largest.

[0100] (III) Prevention and treatment effects on virus-infected Marc-145 cells

[0101] 100TCID 50 PRRSV was inoculated onto Marc-145 cell monolayers at 100 μL / well, incubated at 37°C for 2 h, the virus solution was discarded, and the cells were washed three times with PBS. The traditional Chinese medicine preparations prepared in Example 1 and Comparative Examples 1-4 were added at the maximum cell safety concentration at 100 μL / well, with each dilution replicated in four wells. A blank control group (containing only Marc-145 cell monolayers without other treatments) and a virus control group (Marc-145 cell monolayers inoculated with only 100 TCID45) were also set up. 50 PRRSV (without other treatment) was placed in a 37°C, 5% CO2 incubator for culture. Cell pathogenesis was observed daily, and OD was measured after 7 days. 570nm For specific measurement methods, please refer to (I). See Table 3 for the detailed measurement results.

[0102] Table 3. Prevention and treatment of virus-infected Marc-145 cells by different groups of traditional Chinese medicine preparations.

[0103] Group <![CDATA[OD 570nm Value Blank control group 0.376±0.010 Virus control group 0.230±0.013 Example 1 Group 0.335±0.008 Comparative Example 1 0.251±0.011 Comparative Example 2 0.263±0.010 Comparative Example 3 Groups 0.290±0.009 Comparative Example 4 Groups 0.314±0.012

[0104] As shown in Table 3, the OD of Marc-145 cells using the traditional Chinese medicine preparation from Example 1 group... 570nm The value is relatively the highest, meaning the cell survival is relatively the best, and the effect of preventing and controlling the virus is relatively the strongest.

[0105] (iv) Its role in the prevention and treatment of porcine reproductive and respiratory syndrome.

[0106] Experimental animals were selected based on symptoms including body temperature above 40°C, difficulty breathing (sometimes with abdominal breathing), decreased appetite, diarrhea, emaciation, and motor dysfunction. A total of 140 piglets and adult pigs (both male and female) with PRRS were selected and divided into piglet and adult groups. The piglet group was randomly divided into 7 subgroups of 10 piglets each, including a blank control group, a positive control group, Example 1 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and Comparative Example 4 group. The positive control group received tilmicosin (purchased from Selleck, USA), administered three times daily. Piglets received 5 mL per dose, and adult pigs received 20 mL per dose for 7 consecutive days, fed according to standard methods. Example 1 group and Comparative Examples 1-4 groups received corresponding prepared traditional Chinese medicine preparations, mixed into their feed at a dosage of 1 g / kg / head / day. The blank control group received no other treatment and was fed according to standard methods. The adult pig groups received the same treatment.

[0107] After 14 days of feeding, the treatment situation was observed on the 15th day, and the results are shown in Table 4.

[0108] Efficacy evaluation criteria: Cured pigs have normal body temperature, no diarrhea, normal breathing, no edema, and normal spirit and appetite; Effective pigs have normal body temperature, no diarrhea, mild respiratory symptoms, and normal spirit and appetite; Ineffective pigs do not have their symptoms relieved after medication, or even worsen and die.

[0109] Table 4. Prevention and treatment of PRRS in pigs by different groups of traditional Chinese medicine preparations.

[0110]

[0111] Compared to the blank control group, piglets and adult pigs in the drug treatment group showed varying degrees of relief from clinical symptoms such as depression, loss of appetite, difficulty breathing, and movement disorders as the disease progressed. Group 1 (Example 1) showed the most significant overall relief and the best treatment effect. Among Comparative Examples 1-4, Comparative Example 1 showed a relatively poor overall treatment effect, followed by Comparative Example 2, where the improvement in the mental state of piglets and adult pigs was relatively weaker.

[0112] As can be seen from the above embodiments and experimental examples, the traditional Chinese medicine composition obtained by the various combinations of the present invention can effectively enhance the inhibitory effect on pathogenic microorganisms and antiviral effects, effectively act on PRRSV, improve the immunity of pigs, and enhance the disease resistance of pigs; at the same time, it can alleviate symptoms such as depression and loss of appetite caused by porcine reproductive and respiratory syndrome.

[0113] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome, characterized in that, It is made from the following components in parts by weight: 22-35 parts Indigo Naturalis, 20-26 parts Forsythia suspensa, 13-18 parts Arctium lappa, 18-30 parts Dryopteris crassirhizoma, 16-22 parts Lithospermum erythrorhizon, 10-17 parts Taraxacum mongolicum, 10-15 parts Rheum palmatum, 11-16 parts Scutellaria barbata, 9-14 parts Glycyrrhiza uralensis, 9-13 parts Hordeum vulgare, 10-13 parts Bupleurum chinense, and 7-10 parts Terminalia chebula. The method for preparing the traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome includes the following steps: (1) Take rhubarb, forsythia, dandelion and licorice, cut them into sections and mix them together. Add water and decoct. Filter to obtain filtrate A. (2) Take burdock seed, red-striped gromwell root, dryopteris crassirhizoma, scutellaria barbata, malt, bupleurum root and green fruit, crush them separately and mix them, reflux extract, filter to obtain filtrate B; (3) Mix the obtained filtrate A, filtrate B and indigo and concentrate them to obtain the final product; Step (1) The amount of water added is 3-7 times the total mass of rhubarb, forsythia, dandelion and licorice; the decoction time is 50-80 min / time, and the number of times the decoction is performed is 1-3 times; The solvent used for reflux extraction in step (2) is an ethanol solution with a volume concentration of 65%-85%. The volume mass ratio of the solvent to the total mass of burdock seed, red-striped gromwell root, Dryopteris crassirhizoma, Scutellaria barbata, malt, Bupleurum chinense and green fruit is (8-15) mL:(1-3) g.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, It is made from the following components in parts by weight: 24-32 parts Indigo Naturalis, 21-25 parts Forsythia suspensa, 14-17 parts Arctium lappa, 19-28 parts Dryopteris crassirhizoma, 17-20 parts Lithospermum erythrorhizon, 12-16 parts Taraxacum mongolicum, 11-14 parts Rheum palmatum, 12-15 parts Scutellaria barbata, 10-13 parts Glycyrrhiza uralensis, 10-12 parts Hordeum vulgare, 10.5-12 parts Bupleurum chinense, and 8-9.5 parts Terminalia chebula.

3. A method for preparing the traditional Chinese medicine composition for the prevention and / or treatment of porcine reproductive and respiratory syndrome as described in claim 1 or 2, characterized in that, Includes the following steps: (1) Take rhubarb, forsythia, dandelion and licorice, cut them into sections and mix them together. Add water and decoct. Filter to obtain filtrate A. (2) Take burdock seed, red-striped gromwell root, dryopteris crassirhizoma, scutellaria barbata, malt, bupleurum root and green fruit, crush them separately and mix them, reflux extract, filter to obtain filtrate B; (3) Mix the obtained filtrate A, filtrate B and indigo and concentrate to obtain the final product.

4. The preparation method according to claim 3, characterized in that, Step (1) The amount of water added is 3-7 times the total mass of rhubarb, forsythia, dandelion and licorice; the decoction time is 50-80 min / time, and the number of decoctions is 1-3 times.

5. The preparation method according to claim 3, characterized in that, The solvent used for reflux extraction in step (2) is an ethanol solution with a volume concentration of 65%-85%. The volume mass ratio of the solvent to the total mass of burdock seed, red-striped gromwell root, Dryopteris crassirhizoma, Scutellaria barbata, malt, Bupleurum chinense and green fruit is (8-15) mL:(1-3) g.

6. The preparation method according to claim 3, characterized in that, The reflux extraction time in step (2) is 60-100 min / time, and the number of reflux extractions is 1-4 times.

7. A traditional Chinese medicine preparation for the prevention and / or treatment of porcine reproductive and respiratory syndrome, characterized in that, The traditional Chinese medicine composition comprising the preparation method described in any one of claims 3-6.

8. The traditional Chinese medicine preparation according to claim 7, characterized in that, The dosage forms of the traditional Chinese medicine preparations include oil, powder, granules, or solution.

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