Compound echinacea purpurea extract for treating duck virus hepatitis and preparation method of compound echinacea purpurea extract

Abstract

The invention belongs to the technical field of traditional Chinese veterinary medicines, and relates to a compound echinacea extract for treating duck virus hepatitis and a preparation method of the compound echinacea extract. The compound echinacea purpurea extract is prepared by mixing a traditional Chinese medicine concentrated extract and an echinacea purpurea extract according to a mass ratio of 4: (1-3), the traditional Chinese medicine concentrated extract is prepared from the following raw material medicines: radix bupleuri, radix isatidis, felwort, scutellaria baicalensis, cassia twig, angelica sinensis, oriental wormwood, radix puerariae and pericarpium citri reticulatae. The compound echinacea purpurea extract improves liver injury by improving the autoimmune function of ducks and exerting the antiviral effect of traditional Chinese medicines, and effectively prevents and treats duck viral hepatitis. The components of the composition are all from natural plants, and the composition is free of drug residues and drug resistance after long-term use, obvious in drug effect and small in toxic and side effect.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese veterinary medicines, and particularly relates to a compound echinacea extract for treating duck viral hepatitis and a preparation method thereof. Background Art

[0002] Duck viral hepatitis (DVH) is an acute and severe infectious disease caused by contact, respiratory and digestive tract infection of duck hepatitis virus (DHV). Ducklings are susceptible to it and it has the characteristics of fast transmission speed, wide range and high mortality rate. At present, there are still many problems in the research on the prevention and control of duck viral hepatitis. The failure rate of vaccine immunization is high. Duck viral hepatitis still occurs frequently. It is one of the most important epidemic diseases that endanger the meat duck farming industry in this region, causing huge economic losses.

[0003] Echinacea is a world-famous "immune" herb with a pungent and bitter taste and a cool nature. It enters the lung, spleen, and heart meridians. It has the effects of dispersing wind-heat, clearing away heat and detoxifying, and invigorating qi and strengthening the body. The caffeic acid derivatives, flavonoids, polysaccharides and other substances contained in the extracts of the above-ground parts and roots of Echinacea have significant immune enhancement and antiviral effects. Studies have shown that Echinacea extracts can effectively inhibit the massive reproduction of Newcastle disease virus in chickens in the body, and also have significant inhibitory activity against hepatitis B virus, respiratory syncytial virus, herpes simplex virus and other viruses. Its extracts are widely used at home and abroad, and a variety of preparations have been used in clinical practice. In recent years, many domestic studies have "Chinese medicineized" the exotic plant Echinacea, endowed it with Chinese medicine properties, and made Echinacea better compatible with traditional Chinese medicine and reasonably used in clinical practice.

[0004] As the country pays more and more attention to food safety, the control over the use of various drugs in the livestock and poultry farming industry will become more and more stringent. Therefore, compound Chinese veterinary medicines that are efficient, safe, and not prone to drug resistance have very good market demand and industrialization prospects. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention provides a compound Echinacea extract for treating duck viral hepatitis. The compound Echinacea extract enhances the immune function of ducklings with viral hepatitis, has antioxidant effects, improves liver damage, and has a significant therapeutic effect on duck viral hepatitis.

[0006] To achieve the above purpose, the technical solution adopted by the present invention is: The invention provides a compound echinacea extract for treating duck viral hepatitis. The compound echinacea extract is prepared by mixing a concentrated Chinese medicine extract and an echinacea extract in a mass ratio of 4:1-3.

[0007] The concentrated Chinese medicine extract is prepared from the following raw materials: bupleurum, isatis root, gentian, scutellaria, cassia twig, angelica, capillaris, kudzu root and tangerine peel.

[0008] Bupleurum: bitter in taste, slightly cold in nature, enters the liver and gallbladder meridians, has the effects of reconciling the exterior and interior, soothing the liver and raising yang, and is used for colds and fever, alternating chills and fever, malaria, liver depression and qi stagnation, and chest and rib pain.

[0009] Isatis root: bitter and cold in nature, enters the heart and stomach meridians, clears away heat and detoxifies, cools blood and relieves sore throat, used for wind-heat colds, sore throats, mouth ulcers, and yellow and swollen sores.

[0010] Gentian: bitter in taste, cold in nature, enters the liver and gallbladder meridians, relieves the excess fire of the liver and gallbladder, and eliminates dampness and heat in the lower abdomen. It is mainly used to treat damp-heat jaundice, red and swollen eyes, and itchy eczema.

[0011] Scutellaria baicalensis: bitter in taste, cold in nature, enters the lung, gallbladder, spleen, large intestine and small intestine meridians, clears away heat and dampness, purges fire and detoxifies, stops bleeding, and calms the fetus. It is mainly used to treat lung heat, cough and asthma, gastrointestinal damp-heat, jaundice, high fever and thirst, blood in the stool, epistaxis, red and swollen eyes, carbuncles and furuncles, and fetal movement disorder.

[0012] Cinnamon twig: spicy and sweet in taste, warm in nature, enters the heart, lung and bladder meridians, induces sweat and relaxes muscles, warms and dredges the meridians, assists yang and transforms qi, balances and descends qi, and is used for wind-cold colds, cold pain in the abdomen, amenorrhea due to cold blood, joint pain, phlegm and fluid, edema, palpitations, and running piglet.

[0013] Angelica: Spicy and bitter in taste, warm in nature, enters the heart, liver, and spleen meridians, replenishes blood and promotes blood circulation, regulates menstruation and relieves pain, moisturizes and lubricates the intestines, and is used for irregular menstruation, amenorrhea, dysmenorrhea, accumulation of masses, metrorrhagia, abdominal pain due to deficiency and cold, paralysis, numbness of the skin, dry intestines and difficulty in defecation, heaviness after dysentery, carbuncles, ulcers, and injuries from falls.

[0014] Artemisia capillaris: bitter and pungent in taste, slightly cold in nature, enters the spleen meridian, stomach meridian, and bladder meridian, clears away heat and dampness, promotes bile secretion and reduces jaundice, has the effects of promoting bile secretion, relieving fever, protecting the liver, lowering blood lipids, lowering blood pressure, relieving asthma, inhibiting bacteria, antiviral, diuretic, anti-cancer, analgesic, anti-caries, and anti-inflammatory.

[0015] Pueraria root: sweet and spicy in taste, cool in nature, enters the lung, spleen and stomach meridians, has the functions of relieving muscles and reducing fever, clearing rashes, promoting fluid and quenching thirst, raising yang and stopping diarrhea. It is used for fever due to superficial symptoms, stiffness and pain in the neck and back, measles that does not clear up, thirst due to febrile disease, polydipsia due to yin deficiency, hot diarrhea and dysentery, and diarrhea due to spleen deficiency.

[0016] Tangerine peel: It tastes spicy and bitter, and is warm in nature. It enters the spleen meridian, stomach meridian, and lung meridian. It can regulate qi and strengthen the spleen, regulate the middle, dry up dampness, and resolve phlegm. It is used for abdominal distension or pain and indigestion caused by spleen and stomach qi stagnation.

[0017] In the above prescription, Bupleurum root can resolve the exterior and interior, soothe the liver and raise yang, Isatis root can clear away heat and detoxify, cool blood and reduce swelling to eliminate evil, and the two are used together to clear heat and resist viruses, and are the main medicine; Gentiana scabra can clear the liver and gallbladder fire, remove dampness and heat in the lower part of the body, Scutellaria baicalensis can clear heat and dry dampness, purge fire and detoxify, and Cinnamon twig can warm and dredge the meridians, assist yang and transform qi, and are the minister medicines, which assist the main medicines to play the effect of clearing heat and resisting viruses; Angelica sinensis can replenish blood and activate blood circulation, Artemisia capillaris can clear the liver, promote gallbladder and remove dampness, and Pueraria root can raise yang and stop diarrhea, which are the adjuvant medicines, which can both strengthen the body to eliminate evil and prevent evil from spreading internally; Tangerine peel is the messenger medicine, which harmonizes all the medicines. The whole prescription is reasonably compatible, and all the medicines are used together to play the effects of clearing heat and detoxifying, cooling blood and drying dampness, and replenishing qi and removing blood stasis, which can significantly treat duck viral hepatitis.

[0018] Preferably, the above-mentioned compound echinacea extract for treating duck viral hepatitis, the concentrated Chinese medicine extract, calculated by weight, is made of the following raw materials: 20-30 parts of bupleurum, 20-30 parts of isatis root, 20-30 parts of gentian, 10-20 parts of scutellaria, 10-20 parts of cassia twig, 15-25 parts of angelica, 15-25 parts of capillaris, 10-20 parts of kudzu root and 10-20 parts of tangerine peel.

[0019] More preferably, the above-mentioned compound echinacea extract for treating duck viral hepatitis, the concentrated Chinese medicine extract, calculated by weight, is made of the following raw materials: 25 parts of bupleurum, 25 parts of isatis root, 25 parts of gentian, 15 parts of scutellaria, 15 parts of cinnamon twig, 20 parts of angelica, 20 parts of capillaris, 15 parts of kudzu root and 15 parts of tangerine peel.

[0020] Preferably, the above-mentioned compound echinacea extract for treating duck viral hepatitis, the preparation method of the concentrated Chinese medicine extract is as follows: (1) weighing Radix Isatidis, Gentiana, Scutellaria, Angelica, and Pueraria according to the weight ratio, adding 6-8 times the amount of water, extracting with water for 0.5-1.5h, and filtering the extract; adding 6-8 times the amount of water to the residue to continue extracting for 0.5-1h, filtering the decoction, combining the secondary extracts, and concentrating the obtained extract to obtain a thick paste I; (2) mixing and crushing radix bupleuri, cinnamon twig, capillaris, and dried orange peel, adding 3-8 times the volume of 20-35% ethanol, reflux extracting at 65-75°C for 8-10h, and concentrating the extract to obtain a thick paste II; (3) mixing thick paste I and thick paste II, and spray drying at 60-70°C under vacuum to obtain a concentrated Chinese medicine extract.

[0021] Preferably, the above-mentioned compound Echinacea extract for treating duck viral hepatitis is prepared by the following method: (1) crushing Echinacea, adding 8-10 times the volume of 50-55% ethanol, reflux extraction at 70-80°C for 5-8h, concentrating the extract to obtain a mixture; (2) adding 2-4 times the volume of water to the mixture, then adding ethyl acetate for extraction for 20-25min, and concentrating the extract to dryness under reduced pressure to obtain a crude extract; (3) applying the crude extract to a silica gel column, eluting with a 3:1 ethyl acetate-methanol mixture, collecting the eluate, concentrating, applying to a polyamide column, eluting with 50% ethanol, collecting the eluate, and spray drying at 60-70°C under vacuum to obtain an Echinacea extract.

[0022] The invention provides a Chinese patent medicine of compound echinacea extract for treating duck viral hepatitis, which contains the compound echinacea extract; the dosage form of the Chinese patent medicine is selected from one of powder, tablet, granule, capsule, solution, emulsion and suspension.

[0023] The present invention provides use of the compound echinacea extract for treating duck viral hepatitis in preparing a medicine for treating duck viral hepatitis.

[0024] The present invention conducts syndrome differentiation for duck viral hepatitis. In addition to applying the echinacea extract, the present invention also determines that the medicinal materials such as bupleurum, isatis root, gentian, scutellaria, cinnamon twig, angelica, capillaris, kudzu root and tangerine peel are combined with the echinacea extract to form a prescription to enhance immunity, clear away heat and detoxify, and cool blood and dry dampness.

[0025] Compared with the prior art, the present invention has the following beneficial effects: (1) The compound echinacea extract of the present invention effectively prevents and treats viral hepatitis in ducks by improving the duck's own immune function and exerting the antiviral effect of traditional Chinese medicine. It can also promote growth and improve the body's overall disease resistance.

[0026] (2) The ingredients of the compound echinacea extract of the present invention are all derived from natural plants. Long-term use will result in no drug residue, no drug resistance, significant efficacy and little toxic side effects.

[0027] (3) The present invention is the first to use Echinacea extract in combination with traditional Chinese medicine to treat duck viral hepatitis, making it "traditional Chinese medicine" and giving it the properties and functions of traditional Chinese medicine. It provides experimental data for further appropriate compatibility and rational application of traditional Chinese medicine in the treatment of duck viral hepatitis, and the prospects for application and promotion are very broad. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Comparison of survival rates among groups.

[0029] Figure 2 Comparison of AST in each group, compared with group B, * P<0.05,** P<0.01; compared with group D, # P<0.05.

[0030] Figure 3 Comparison of ALT in each group, compared with group B, * P<0.05, ** P<0.01; compared with group D, # P<0.05.

[0031] Figure 4 Comparison of GSH in each group, compared with group B, * P<0.05, ** P<0.01; compared with group D, # P<0.05.

[0032] Figure 5 Comparison of SOD in each group, compared with group B, * P<0.05, ** P<0.01; compared with group D, # P<0.05.

[0033] Figure 6 Comparison of MDA in each group, compared with group B, * P<0.05, ** P<0.01; compared with group D, # P<0.05.

[0034] Figure 7 Comparison of spleen index in each group, compared with group B, * P<0.05, ** P<0.01.

[0035] Figure 8 Comparison of thymus index in each group, compared with group B, * P<0.05, ** P<0.01.

[0036] Fig. 9 Comparison of bursal index of each group, compared with group B, * P<0.05, ** P<0.01. DETAILED DESCRIPTION

[0037] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are 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 those skilled in the art without creative work are within the scope of protection of the present invention.

[0038] Example 1 A compound echinacea extract for treating duck viral hepatitis is prepared by mixing a concentrated Chinese medicine extract and an echinacea extract in a mass ratio of 4:2.

[0039] The concentrated Chinese medicine extract is prepared from the following raw materials by weight: 25 parts of bupleurum, 25 parts of isatis root, 25 parts of gentian, 15 parts of scutellaria, 15 parts of cassia twig, 20 parts of angelica, 20 parts of capillaris, 15 parts of kudzu root and 15 parts of dried tangerine peel.

[0040] The preparation method of the concentrated Chinese medicine extract is as follows: (1) Weigh Radix Isatidis, Radix Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, add 6 times the amount of water, extract with water for 1 hour, and filter the extract; add 6 times the amount of water to the residue and continue to extract for 0.5 hour, filter the decoction, combine the secondary extracts, and concentrate the obtained extract to obtain thick paste I; (2) Mix and crush Bupleurum, Cassia Twig, Artemisia Capillaris, and Tangerine Peel, add 5 times the volume of 30% ethanol, reflux at 70°C for 8 hours, and concentrate the extract to obtain thick paste II; (3) Mix thick paste I and thick paste II, and spray dry them at 60°C under vacuum to obtain a concentrated extract of traditional Chinese medicine.

[0041] The preparation method of the Echinacea extract is as follows: (1) Crush Echinacea, add 8 times the volume of 50% ethanol, reflux extract at 75°C for 6 hours, and concentrate the extract to obtain a mixture; (2) Add 3 times the volume of water to the mixture, then add ethyl acetate to extract for 20 minutes, and concentrate the extract to dryness under reduced pressure to obtain a crude extract; (3) Apply the crude extract to a silica gel column, elute with a 3:1 ethyl acetate-methanol mixture, collect the eluate, concentrate, apply to a polyamide column, elute with 50% ethanol, collect the eluate, and spray dry at 70°C under vacuum to obtain an Echinacea extract.

[0042] Example 2 A compound echinacea extract for treating duck viral hepatitis is prepared by mixing a concentrated Chinese medicine extract and an echinacea extract in a mass ratio of 4:1.

[0043] The concentrated Chinese medicine extract is prepared from the following raw materials by weight: 30 parts of bupleurum, 30 parts of isatis root, 20 parts of gentian, 20 parts of scutellaria, 10 parts of cassia twig, 25 parts of angelica, 15 parts of capillaris, 20 parts of kudzu root and 20 parts of tangerine peel.

[0044] The preparation method of the concentrated Chinese medicine extract is as follows: (1) Weigh Radix Isatidis, Radix Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, add 8 times the amount of water, extract with water for 1.5 hours, and filter the extract; add 8 times the amount of water to the residue and continue to extract for 1 hour, filter the decoction, combine the secondary extracts, and concentrate the obtained extract to obtain thick paste I; (2) Mix and crush Bupleurum, Cassia Twig, Artemisia Capillaris, and Tangerine Peel, add 8 times the volume of 35% ethanol, reflux at 75°C for 10 hours, and concentrate the extract to obtain thick paste II; (3) Mix thick paste I and thick paste II, and spray dry them at 70°C under vacuum to obtain a concentrated extract of traditional Chinese medicine.

[0045] The preparation method of the Echinacea extract is as follows: (1) Crush Echinacea, add 10 times the volume of 55% ethanol, reflux extract at 70°C for 8 hours, and concentrate the extract to obtain a mixture; (2) Add 4 times the volume of water to the mixture, then add ethyl acetate to extract for 25 minutes, and concentrate the extract to dryness under reduced pressure to obtain a crude extract; (3) Apply the crude extract to a silica gel column, elute with a 3:1 ethyl acetate-methanol mixture, collect the eluate, concentrate, apply to a polyamide column, elute with 50% ethanol, collect the eluate, and spray dry at 70°C under vacuum to obtain an Echinacea extract.

[0046] Example 3 A compound echinacea extract for treating duck viral hepatitis is prepared by mixing a concentrated Chinese medicine extract and an echinacea extract in a mass ratio of 4:3.

[0047] The concentrated Chinese medicine extract is prepared from the following raw materials by weight: 20 parts of bupleurum, 20 parts of isatis root, 30 parts of gentian, 10 parts of scutellaria, 20 parts of cassia twig, 15 parts of angelica, 25 parts of capillaris, 10 parts of kudzu root and 10 parts of tangerine peel.

[0048] The preparation method of the concentrated Chinese medicine extract is as follows: (1) Weigh Radix Isatidis, Radix Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, add 7 times the amount of water, extract with water for 0.5 h, and filter the extract; add 7 times the amount of water to the residue and continue to extract for 0.5 h, filter the decoction, combine the secondary extracts, and concentrate the obtained extract to obtain thick paste I; (2) Mix and crush Bupleurum, Cassia Twig, Artemisia Capillaris, and Tangerine Peel, add 3 times the volume of 25% ethanol, reflux at 65°C for 8 h, and concentrate the extract to obtain thick paste II; (3) Mix thick paste I and thick paste II, and spray dry them at 60°C under vacuum to obtain a concentrated extract of traditional Chinese medicine.

[0049] The preparation method of the Echinacea extract is as follows: (1) Crush Echinacea, add 8 times the volume of 50% ethanol, reflux extract at 70°C for 5 hours, and concentrate the extract to obtain a mixture; (2) Add 2 times the volume of water to the mixture, then add ethyl acetate to extract for 25 minutes, and concentrate the extract to dryness under reduced pressure to obtain a crude extract; (3) Apply the crude extract to a silica gel column, elute with a 3:1 ethyl acetate-methanol mixture, collect the eluate, concentrate, apply to a polyamide column, elute with 50% ethanol, collect the eluate, and spray dry at 60°C under vacuum to obtain an Echinacea extract.

[0050] Example 4 A compound echinacea extract for treating duck viral hepatitis is prepared by mixing a concentrated Chinese medicine extract and an echinacea extract in a mass ratio of 4:2.

[0051] The concentrated Chinese medicine extract is prepared from the following raw materials by weight: 25 parts of bupleurum, 30 parts of isatis root, 20 parts of gentian, 10 parts of scutellaria, 20 parts of cassia twig, 20 parts of angelica, 20 parts of capillaris, 15 parts of kudzu root and 20 parts of tangerine peel.

[0052] The preparation method of the concentrated Chinese medicine extract is as follows: (1) Weigh Radix Isatidis, Herba Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, add 6 times the amount of water, extract with water for 0.5 h, and filter the extract; add 6 times the amount of water to the residue and continue to extract for 0.5 h, filter the decoction, combine the secondary extracts, and concentrate the obtained extracts to obtain thick paste I; (2) Mix and crush Bupleurum, Cinnamon Twig, Artemisia Capillaris, and Citrus Reticulatae, add 3 times the volume of 20% ethanol, reflux at 65°C for 8 h, and concentrate the extract to obtain thick paste II; (3) Mix thick paste I and thick paste II, and spray dry them at 60°C under vacuum to obtain a concentrated extract of traditional Chinese medicine.

[0053] The preparation method of the Echinacea extract is as follows: (1) Crush Echinacea, add 8 times the volume of 50% ethanol, reflux extract at 70°C for 5 hours, and concentrate the extract to obtain a mixture; (2) Add 2 times the volume of water to the mixture, then add ethyl acetate to extract for 20 minutes, and concentrate the extract to dryness under reduced pressure to obtain a crude extract; (3) Apply the crude extract to a silica gel column, elute with a 3:1 ethyl acetate-methanol mixture, collect the eluate, concentrate, apply to a polyamide column, elute with 50% ethanol, collect the eluate, and spray dry at 60°C under vacuum to obtain an Echinacea extract.

[0054] Example 5 A compound echinacea extract for treating duck viral hepatitis is prepared by mixing a concentrated Chinese medicine extract and an echinacea extract in a mass ratio of 4:3.

[0055] The concentrated Chinese medicine extract is prepared from the following raw materials by weight: 30 parts of bupleurum, 20 parts of isatis root, 25 parts of gentian, 15 parts of scutellaria, 20 parts of cassia twig, 15 parts of angelica, 20 parts of capillaris, 15 parts of kudzu root and 10 parts of tangerine peel.

[0056] The preparation method of the concentrated Chinese medicine extract is as follows: (1) Weigh Radix Isatidis, Herba Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, add 7 times the amount of water, extract with water for 1 hour, and filter the extract; add 7 times the amount of water to the residue and continue to extract for 0.5 hour, filter the decoction, combine the secondary extracts, and concentrate the obtained extract to obtain thick paste I; (2) Mix and crush Bupleurum, Cinnamon Twig, Artemisia Capillaris, and Tangerine Peel, add 6 times the volume of 25% ethanol, reflux at 70°C for 9 hours, and concentrate the extract to obtain thick paste II; (3) Mix thick paste I and thick paste II, and spray dry them at 65°C under vacuum to obtain a concentrated extract of traditional Chinese medicine.

[0057] The preparation method of the Echinacea extract is as follows: (1) Crush Echinacea, add 9 times the volume of 50% ethanol, reflux extract at 75°C for 7 hours, and concentrate the extract to obtain a mixture; (2) Add 3 times the volume of water to the mixture, then add ethyl acetate to extract for 22 minutes, and concentrate the extract to dryness under reduced pressure to obtain a crude extract; (3) Apply the crude extract to a silica gel column, elute with a 3:1 ethyl acetate-methanol mixture, collect the eluate, concentrate, apply to a polyamide column, elute with 50% ethanol, collect the eluate, and spray dry at 65°C under vacuum to obtain an Echinacea extract.

[0058] Example 6 A compound echinacea extract for treating duck viral hepatitis is prepared by mixing a concentrated Chinese medicine extract and an echinacea extract in a mass ratio of 4:1.

[0059] The concentrated Chinese medicine extract is prepared from the following raw materials by weight: 25 parts of bupleurum, 25 parts of isatis root, 20 parts of gentian, 20 parts of scutellaria, 10 parts of cassia twig, 20 parts of angelica, 15 parts of capillaris, 10 parts of kudzu root and 20 parts of dried tangerine peel.

[0060] The preparation method of the concentrated Chinese medicine extract is as follows: (1) Weigh Radix Isatidis, Radix Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, add 6 times the amount of water, extract with water for 1.5 hours, and filter the extract; add 6 times the amount of water to the residue and continue to extract for 0.5 hours, filter the decoction, combine the secondary extracts, and concentrate the obtained extracts to obtain thick paste I; (2) Mix and crush Bupleurum, Cassia Twig, Artemisia Capillaris, and Tangerine Peel, add 8 times the volume of 35% ethanol, reflux at 75°C for 10 hours, and concentrate the extract to obtain thick paste II; (3) Mix thick paste I and thick paste II, and spray dry them at 70°C under vacuum to obtain a concentrated extract of traditional Chinese medicine.

[0061] The preparation method of the Echinacea extract is as follows: (1) Crush Echinacea, add 10 times the volume of 55% ethanol, reflux extract at 80°C for 8 hours, and concentrate the extract to obtain a mixture; (2) Add 3 times the volume of water to the mixture, then add ethyl acetate to extract for 25 minutes, and concentrate the extract to dryness under reduced pressure to obtain a crude extract; (3) Apply the crude extract to a silica gel column, elute with a 3:1 ethyl acetate-methanol mixture, collect the eluate, concentrate, apply to a polyamide column, elute with 50% ethanol, collect the eluate, and spray dry at 65°C under vacuum to obtain an Echinacea extract.

[0062] Comparative Example 1 Compared with Example 1, the difference is that the Echinacea extract is not contained.

[0063] Comparative Example 2 Compared with Example 1, the difference is that the concentrated extract of traditional Chinese medicine is not contained.

[0064] Efficacy Example 1 Study on the effect of the Echinacea extract of the present invention on duck viral hepatitis 1. Experimental Methods 240 one-day-old non-immunized ducks were randomly divided into 6 groups when they were raised to 4-6 days of age, with 40 ducks in each group, a blank control group (raised alone, recorded as Group A), a model control group (recorded as Group B), a western medicine control group (recorded as Group C), Example 1 group (recorded as Group D), Comparative Example 1 group (recorded as Group E), and Comparative Example 2 group (recorded as Group F).

[0065] Each of the B, C, D, E, and F groups was injected intramuscularly with 0.2 mL of DHAV-1 virus, and the same volume of saline was injected into the A group. After the challenge, the C group was given 1.25 mg of ribavirin solution, and the D, E, and F groups were given 5 mg / mouse / day of the drug mixed in drinking water for 3 consecutive days. The B group was given saline instead of the drug.

[0066] 8h and 54h after the challenge, 5 ducklings were randomly picked from each group, and 5g of liver tissue was taken and stored in a cryotube in liquid nitrogen for the detection of antioxidant enzyme activity. Clinical symptoms were observed regularly every day and mortality was recorded for 5 consecutive days. Whenever a duckling died, it was immediately dissected and observed for significant pathological changes.

[0067] 2. Determination of indicators 2.1 Survival rate When calculating the final survival rate of each group, the sample was removed (a total of 10 animals in each group).

[0068] Survival rate (%) = number of survivors / number of valid samples in each group × 100%.

[0069] 2.2 Biochemical markers of liver damage Serum aspartate aminotransferase (ALT) and alanine aminotransferase (AST) were measured.

[0070] 2.3 Indicators of liver peroxidation damage The kit was used to detect GSH, MDA and SOD in liver tissue.

[0071] 3. Statistical methods use The software processes the data and measures the data. The comparison among multiple groups was performed by one-way analysis of variance, and the comparison among groups was performed by Pairwise comparisons were performed, and P < 0.05 was considered statistically significant.

[0072] 4. Experimental Results 4.1 Comparison of survival rates among groups 28 rats survived in group D, with a survival rate of 93.3%. The survival rate of group D was significantly higher than that of groups C, E, and F, indicating that group D had the best treatment effect. Figure 1 .

[0073] The compound echinacea extract of the invention has a good therapeutic effect on duck viral hepatitis.

[0074] 4.2 Comparison of biochemical indicators of liver injury among groups AST and ALT in group B were significantly higher than those in group A. AST and ALT in group D were significantly lower than those in group B, and the levels of each index were basically close to those in group A. The effect of improving AST and ALT levels in group D was better than that in groups C, E, and F. Figure 2 , 3 .

[0075] The compound echinacea extract of the present invention can effectively alleviate the liver function damage caused by duck viral hepatitis infection.

[0076] 4.3 Comparison of liver peroxidation damage indicators among groups Compared with group A, the contents of SOD and GSH in liver homogenate of group B were significantly decreased, and the content of MDA was significantly increased.

[0077] Compared with group B, each drug-treated group could increase the content of SOD and GSH in liver homogenate and reduce the content of MDA. Group D was better than group C, E and F in improving the content of SOD, GSH and MDA. Figure 4-6 .

[0078] The compound echinacea extract of the present invention can effectively alleviate liver damage caused by duck viral hepatitis infection.

[0079] Efficacy Example 2 Study on the effect of the Echinacea extract of the present invention on the immune function of duck viral hepatitis 1. Experimental Methods 80 one-day-old healthy Cherry Valley ducks were randomly divided into 4 groups when they were raised to 4-6 days of age, with 20 ducks in each group, a blank control group (raised alone, recorded as Group A), a model control group (recorded as Group B), a western medicine control group (recorded as Group C), and Example 1 group (recorded as Group D).

[0080] Each of the B, C, and D groups was injected intramuscularly with 0.2 mL of DHAV-1 virus, and the same volume of saline was injected into the A group. After the challenge, the C group was given 1.25 mg of ribavirin solution, and the D group was given 5 mg / head / day of the drug mixed in drinking water for 3 consecutive days. The B group was given saline instead of the drug. The experimental period was 30 days.

[0081] 2. Determination of indicators Immune organ index Experiment 6: Body weight was measured on day 30 (fasting for 12 h). Spleen, bursa of Fabricius index, and thymus were collected after bleeding from the neck and slaughtered. The surface was cleaned with filter paper, and the mass was weighed with a precision electronic balance and recorded (mg).

[0082] Immune organ index / (mg / g) = immune organ mass / broiler duck body mass.

[0083] 3. Statistical methods use The software processes the data and measures the data. The comparison among multiple groups was performed by one-way analysis of variance, and the comparison among groups was performed by Pairwise comparisons were performed, and P < 0.05 was considered statistically significant.

[0084] 4. Experimental Results Comparison of immune organ index Compared with group A, the spleen index, thymus index and bursa index of group B were significantly reduced.

[0085] Compared with group B, each drug-treated group can promote spleen index, thymus index and bursal index. Figure 7-9 .

[0086] The compound echinacea extract of the invention has a tendency to promote spleen index, thymus index and bursa index, and enhances the immune function of ducklings with viral hepatitis.

[0087] The above embodiments are merely examples for clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description, and the obvious changes or modifications derived therefrom are still within the scope of protection of the present invention.

Claims

1. A compound Echinacea extract for treating duck viral hepatitis, characterized in that: It is prepared by mixing concentrated extracts of traditional Chinese medicine and Echinacea extract in a mass ratio of 4:1-3; The concentrated Chinese medicine extract is prepared from the following raw materials in parts by weight: 20-30 parts of bupleurum, 20-30 parts of isatis root, 20-30 parts of gentian, 10-20 parts of scutellaria, 10-20 parts of cassia twig, 15-25 parts of angelica, 15-25 parts of capillaris, 10-20 parts of kudzu root and 10-20 parts of tangerine peel.

2. The compound Echinacea extract for treating duck viral hepatitis according to claim 1, characterized in that: The concentrated Chinese medicine extract is prepared from the following raw materials in parts by weight: 25 parts of bupleurum, 25 parts of isatis root, 25 parts of gentian, 15 parts of scutellaria, 15 parts of cassia twig, 20 parts of angelica, 20 parts of capillaris, 15 parts of kudzu root and 15 parts of tangerine peel.

3. The compound Echinacea extract for treating duck viral hepatitis according to claim 1, characterized in that: The preparation method of the concentrated Chinese medicine extract is as follows: (1) weighing Radix Isatidis, Radix Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, adding 6-8 times the amount of water, extracting with water for 0.5-1.5 hours, and filtering the extract; adding 6-8 times the amount of water to the residue to continue extracting for 0.5-1 hours, filtering the decoction, combining the secondary extracts, and concentrating the obtained extract to obtain a thick paste I; (2) mixing and crushing radix bupleuri, cassia twig, artemisia capillaris, and tangerine peel, adding 3-8 times the volume of 20-35% ethanol, refluxing at 65-75° C. for extraction for 8-10 hours, and concentrating the extract to obtain a thick paste II; (3) mixing the thick paste I and the thick paste II, and spray drying at 60-70° C. under vacuum to obtain a concentrated Chinese medicine extract.

4. The compound Echinacea extract for treating duck viral hepatitis according to claim 3, characterized in that: The preparation method of the concentrated Chinese medicine extract is as follows: (1) weighing Radix Isatidis, Radix Gentianae, Radix Scutellariae, Radix Angelicae Sinensis, and Radix Puerariae according to the weight ratio, adding 6 times the amount of water, extracting with water for 1 hour, and filtering the extract; adding 6 times the amount of water to the residue to continue extracting for 0.5 hours, filtering the decoction, combining the secondary extracts, and concentrating the obtained extract to obtain a thick paste I; (2) mixing and crushing radix bupleuri, cassia twig, artemisia capillaris, and tangerine peel, adding 5 times the volume of 30% ethanol, refluxing at 70°C for extraction for 8 hours, and concentrating the extract to obtain a thick paste II; (3) mixing the thick paste I and the thick paste II, and spray drying at 60°C under vacuum to obtain a concentrated Chinese medicine extract.

5. The compound Echinacea extract for treating duck viral hepatitis according to claim 1, characterized in that: The preparation method of the Echinacea extract is as follows: (1) crushing Echinacea, adding 8-10 times the volume of 50-55% ethanol, refluxing extraction at 70-80°C for 5-8h, concentrating the extract to obtain a mixture; (2) adding 2-4 times the volume of water to the mixture, then adding ethyl acetate for extraction for 20-25min, and concentrating the extract to dryness under reduced pressure to obtain a crude extract; (3) applying the crude extract to a silica gel column, eluting with a 3:1 ethyl acetate-methanol mixture, collecting the eluate, concentrating the crude extract, applying the crude extract to a polyamide column, eluting with 50% ethanol, collecting the eluate, and vacuum low-temperature spray drying at 60-70°C to obtain the Echinacea extract.

6. The compound Echinacea extract for treating duck viral hepatitis according to claim 5, characterized in that: The preparation method of the Echinacea extract is as follows: (1) crushing Echinacea, adding 8 times the volume of 50% ethanol, refluxing extraction at 75°C for 6 hours, concentrating the extract to obtain a mixture; (2) adding 3 times the volume of water to the mixture, then adding ethyl acetate to extract for 20 minutes, and concentrating the extract to dryness under reduced pressure to obtain a crude extract; (3) applying the crude extract to a silica gel column, eluting with a 3:1 ethyl acetate-methanol mixture, collecting the eluate, concentrating the crude extract, applying the crude extract to a polyamide column, eluting with 50% ethanol, collecting the eluate, and spray drying at 70°C under vacuum to obtain the Echinacea extract.

7. A Chinese patent medicine of compound Echinacea extract for treating duck viral hepatitis, characterized in that: The invention is prepared from the compound echinacea extract according to any one of claims 1 to 6.

8. The Chinese patent medicine of compound Echinacea extract for treating duck viral hepatitis according to claim 7, characterized in that: The dosage form of the Chinese patent medicine is selected from one of powder, tablet, granule, capsule, solution, emulsion and suspension.

9. Use of the compound Echinacea extract for treating duck viral hepatitis according to any one of claims 1 to 6 in the preparation of a drug for treating duck viral hepatitis.

Images