A traditional Chinese medicine composition for the prevention, relief or treatment of fatty liver in fish and its application.

Abstract

This invention provides a traditional Chinese medicine composition for the prevention, relief, or treatment of fatty liver in fish and its application, belonging to the field of veterinary products. The active ingredients of the traditional Chinese medicine composition include Terminalia chebula, and also include one or more of Galla chinensis, Salvia miltiorrhiza, Schisandra chinensis, Glycyrrhiza uralensis, Eucommia ulmoides, and Paeonia lactiflora. The traditional Chinese medicine composition of this invention has the effects of protecting the liver, enhancing immunity, improving blood circulation, and regulating lipid metabolism, and is safe and residue-free.

Description

Technical Field

[0001] This invention relates to the field of veterinary medicine technology, specifically to a traditional Chinese medicine composition for the prevention, relief or treatment of fatty liver or liver damage in fish, its preparation method and application. Background Technology

[0002] California bass (Micropterus salmoides) is an important economic fish species, favored by fish farmers for its rapid growth and delicious flesh. In recent years, with the promotion of high-density intensive aquaculture, the scale of California bass farming has been continuously expanding. However, in high-density farming environments, California bass are susceptible to various diseases, with fatty liver and liver damage being the most common. These diseases not only affect the fish's growth rate and feed conversion rate but also lead to decreased immunity, increasing the risk of secondary infections, and in severe cases, even causing large-scale mortality, resulting in huge economic losses for fish farmers.

[0003] The occurrence of fatty liver and liver damage in California bass is related to multiple factors, mainly including: 1) Feed factors: High-fat diets or nutritionally unbalanced diets can lead to excessive fat accumulation in the liver, causing fatty liver; 2) Environmental factors: Environmental stresses such as deteriorating water quality, insufficient dissolved oxygen, and excessive ammonia nitrogen and nitrite levels can increase the burden on the liver, leading to liver damage; 3) Drug abuse: Long-term use of antibiotics or chemical drugs to prevent and treat diseases may cause secondary damage to the liver; 4) Metabolic disorders: Under high-density farming conditions, the metabolic load of fish increases, which can easily lead to glucose and lipid metabolism disorders, further aggravating liver lesions.

[0004] Currently, the prevention and treatment of fatty liver and liver damage in California bass mainly rely on antibiotics and chemical drugs. However, long-term use can easily lead to problems such as drug residues, bacterial resistance, and reduced metabolism, endangering food safety and the ecological environment. Therefore, developing safe, effective, and residue-free traditional Chinese medicine additives is of significant practical importance. Summary of the Invention

[0005] Purpose of the invention

[0006] The purpose of this invention is to provide a traditional Chinese medicine composition for the prevention, relief, or treatment of fatty liver in fish and its application. The traditional Chinese medicine composition of this invention has the effects of protecting the liver, enhancing immunity, improving blood circulation, and regulating lipid metabolism, and is safe and residue-free.

[0007] Solution

[0008] To achieve the objectives of this invention, the technical solution adopted is as follows:

[0009] In a first aspect, the present invention provides a traditional Chinese medicine composition for preventing, alleviating or treating fatty liver in fish and its application, wherein the active ingredients of the traditional Chinese medicine composition include green fruit, and also include one or more of gallnut, salvia miltiorrhiza, schisandra chinensis, licorice, eucommia ulmoides and white peony root.

[0010] Furthermore, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula and 10-20 parts of Galla chinensis;

[0011] Alternatively, the weight ratio of green fruit to gallnut is (40~80):(10~20), (2~16):1, (2~6):1, 2:1, 4:1, 6:1 or 10:3.

[0012] Furthermore, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 0-20 parts of Galla chinensis, 0-10 parts of Salvia miltiorrhiza, 0-15 parts of Schisandra chinensis, 0-10 parts of Glycyrrhiza uralensis, 0-10 parts of Eucommia ulmoides, and 0-10 parts of Paeonia lactiflora.

[0013] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 0-10 parts of Salvia miltiorrhiza, 0-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0014] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0015] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0016] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5 parts of Paeonia lactiflora.

[0017] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 0-20 parts of Galla chinensis, 0-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0018] Furthermore, the weight fraction of green fruit in the traditional Chinese medicine composition is more than 35%, optionally ≥40%, optionally 40%~80%, optionally within any two values ​​of 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%.

[0019] Furthermore, the dosage form of the traditional Chinese medicine composition is any one of powder, oral liquid, spray, ointment, injection, and capsule;

[0020] And / or, the fish is a bass, optionally a California bass;

[0021] And / or, the product is a drug, feed, or feed additive; optionally, the feed is fish feed.

[0022] Furthermore, the product is obtained by extracting traditional Chinese medicine; optionally, the product is an aqueous extract, or optionally, a hydrothermal extract.

[0023] Optionally, the product is an extract obtained by pulverizing and decocting in water;

[0024] Optionally, the product is an extract obtained by decocting various medicinal powders with 5 to 15 times the amount of water, and optionally, the extract is decocted, concentrated, and dried to obtain powder; optionally, the decoction process involves 1 to 3 filtrations, and the residue is decocted again; optionally, the concentration method is vacuum concentration to a relative density of 1.10 to 1.20, optionally at a temperature of 60°C; the vacuum-concentrated extract is dried and pulverized to obtain a traditional Chinese medicine additive powder;

[0025] Optionally, the preparation method of the product includes: washing the medicinal materials, drying them at 50-60℃, pulverizing them, passing them through a 60-80 mesh sieve, mixing them evenly, adding 5-15 times the amount of purified water to the mixed Chinese medicine powder, heating it to boiling and decocting it for 1-3 hours, filtering it to obtain the first decoction, adding 5-10 times the amount of water to the dregs again, decocting it for 1-2 hours, filtering it to obtain the second decoction, and combining the two decoctions; concentrating the decoction under reduced pressure at 60℃ to a relative density of 1.10-1.20 (measured at 60℃) to obtain an extract; drying the extract at 60-70℃ until the moisture content is less than 5% to obtain a dry extract, then pulverizing it and passing it through a 60-80 mesh sieve to obtain a uniform Chinese medicine additive powder.

[0026] Secondly, a traditional Chinese medicine composition for preventing, alleviating or treating fatty liver in fish is provided, wherein the active ingredients of the traditional Chinese medicine composition include Terminalia chebula, and also include one or more of Galla chinensis, Salvia miltiorrhiza, Schisandra chinensis, Glycyrrhiza uralensis, Eucommia ulmoides and Paeonia lactiflora.

[0027] Furthermore, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 0-20 parts of Galla chinensis, 0-10 parts of Salvia miltiorrhiza, 0-15 parts of Schisandra chinensis, 0-10 parts of Glycyrrhiza uralensis, 0-10 parts of Eucommia ulmoides, and 0-10 parts of Paeonia lactiflora.

[0028] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 0-10 parts of Salvia miltiorrhiza, 0-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0029] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0030] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0031] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Galla chinensis, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5 parts of Paeonia lactiflora.

[0032] Alternatively, the weight ratio of green fruit to gallnut is (40~80):(10~20), (2~16):1, (2~6):1, 2:1, 4:1, 6:1 or 10:3.

[0033] Optionally, the active ingredients of the traditional Chinese medicine composition include the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 0-20 parts of Galla chinensis, 0-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

[0034] Optionally, the weight fraction of green fruit in the traditional Chinese medicine composition is more than 35%, optionally ≥40%, optionally 40%~80%, optionally within any two values ​​of 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%;

[0035] Optionally, the dosage form of the traditional Chinese medicine composition is any one of powder, oral liquid, spray, ointment, injection, and capsule;

[0036] Optionally, the fish is a bass, and optionally a California bass;

[0037] Optionally, the product is a drug, feed, or feed additive; optionally, the feed is fish feed.

[0038] Thirdly, a method for preparing the traditional Chinese medicine composition described in the second aspect is provided, comprising the following steps: obtaining the traditional Chinese medicine composition by extraction; optionally, the product is an aqueous extract, optionally, a hydrothermal extract;

[0039] Optionally, the product is an extract obtained by pulverizing and decocting in water;

[0040] Optionally, the product is an extract obtained by decocting various medicinal powders with 5 to 15 times the amount of water, and optionally, the extract is decocted, concentrated, and dried to obtain powder; optionally, the decoction process involves 1 to 3 filtrations, and the residue is decocted again; optionally, the concentration method is vacuum concentration to a relative density of 1.10 to 1.20, optionally at a temperature of 60°C; the vacuum-concentrated extract is dried and pulverized to obtain a traditional Chinese medicine additive powder;

[0041] Optionally, the preparation method of the product includes: washing the medicinal materials, drying them at 50-60℃, pulverizing them, passing them through a 60-80 mesh sieve, mixing them evenly, adding 5-15 times the amount of purified water to the mixed Chinese medicine powder, heating it to boiling and decocting it for 1-3 hours, filtering it to obtain the first decoction, adding 5-10 times the amount of water to the dregs again, decocting it for 1-2 hours, filtering it to obtain the second decoction, and combining the two decoctions; concentrating the decoction under reduced pressure at 60℃ to a relative density of 1.10-1.20 (measured at 60℃) to obtain an extract; drying the extract at 60-70℃ until the moisture content is less than 5% to obtain a dry extract, then pulverizing it and passing it through a 60-80 mesh sieve to obtain a uniform Chinese medicine additive powder.

[0042] Fourthly, a fish feed is provided, comprising a basic fish feed, and further comprising the traditional Chinese medicine composition described in the second aspect, or the product prepared by the preparation method described in the third aspect, wherein optionally, the composition accounts for 1-10‰ of the feed by weight, or optionally 6-10‰; optionally, the basic fish feed is a bass basic feed.

[0043] Beneficial effects

[0044] The traditional Chinese medicine composition of this invention has the effects of protecting the liver, enhancing immunity, improving blood circulation, and regulating lipid metabolism, and is safe and residue-free. This composition can effectively improve liver function in California bass, repair liver cell damage, effectively reduce liver fat content, promote toxin excretion, and enhance the immunity and disease resistance of California bass. It can also promote liver blood flow, repair damaged liver cells, and alleviate liver enlargement or necrosis caused by hypoxia or toxin accumulation in aquatic animals. It can prevent fatty liver problems caused by high-fat diets in aquatic animals by inhibiting fat deposition in the liver. The traditional Chinese medicine additive of this invention uses pure natural Chinese medicinal materials, is safe and non-toxic, and leaves no drug residues, meeting the requirements of green aquaculture. The traditional Chinese medicine composition of this invention also has effects such as protecting the liver, optimizing intestinal flora, promoting fat absorption, anti-inflammation, and degrading aflatoxin and antibiotics, further protecting liver function. Attached Figure Description

[0045] One or more embodiments are illustrated by way of example with reference to the accompanying drawings, and these illustrative examples are not intended to limit the embodiments. The term "illustrative" as used herein means "serving as an example, embodiment, or illustration." Any embodiment illustrated herein as "illustrative" is not necessarily to be construed as superior to or better than other embodiments.

[0046] Figure 1 Anatomical images of fish corresponding to the liver damage symptom score of this invention.

[0047] Figure 2 Photographs of the screening of antioxidant Chinese medicinal materials in Experiment Example 1 of this invention. Detailed Implementation

[0048] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0049] Furthermore, to better illustrate the present invention, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In some embodiments, materials, elements, methods, and means well known to those skilled in the art are not described in detail in order to highlight the spirit of the invention.

[0050] In the herbal composition of this invention, herbs such as Terminalia chebula and Galla chinensis are used to protect the liver, effectively improving liver function and repairing liver cell damage in California bass. Herbs such as Salvia miltiorrhiza and Schisandra chinensis can lower lipids and detoxify, effectively reducing the fat content in the liver of California bass and promoting toxin excretion. Herbs such as Glycyrrhiza uralensis can improve liver blood circulation, enhance the immunity of California bass, and improve its disease resistance. Herbs such as Paeonia lactiflora promote liver blood flow, repair damaged liver cells, and alleviate liver enlargement or necrosis caused by hypoxia or toxin accumulation in aquatic animals. Terminalia chebula and Eucommia ulmoides can regulate lipid metabolism and have a lipid-lowering effect, potentially preventing fatty liver problems in aquatic animals caused by high-fat feed by inhibiting fat deposition in the liver. The herbal additives of this invention use pure natural herbal raw materials, are safe and non-toxic, have no drug residues, and meet the requirements of green aquaculture.

[0051] All the Chinese medicinal materials used in this invention can be obtained commercially.

[0052] In the following experiments, the liver injury symptom scoring method in Table 1 was used, as shown in the reference images. Figure 1 .

[0053] Table 1. Liver Injury Symptom Scores

[0054] .

[0055] In the following experiments, the California bass feed used was a conventional feed. Optionally, the feed composition used in the following experiments is shown in Table 2, and the nutrient content is shown in Table 3.

[0056] Table 2. Feed Composition

[0057] ;

[0058] Table 3. Nutrient content of feed (%) is as follows (actual measured values ​​DM)

[0059] ;

[0060] The preparation methods of the traditional Chinese medicine additives in the following examples and comparative examples include the following steps:

[0061] Raw material pretreatment:

[0062] Clean the Chinese medicinal herbs according to the specified proportions, remove impurities, and then dry them at 50-60℃ until the moisture content is below 10%. Crush the dried raw materials separately and pass them through a 60-80 mesh sieve to obtain uniform powder.

[0063] Mixing and decoction: Add 10 times the amount of purified water to the mixed herbal powder, heat to boiling, and decoct for 2 hours. Filter to obtain the first decoction. Add 8 times the amount of water to the dregs again, decoct for 1.5 hours, and filter to obtain the second decoction. Combine the two decoctions.

[0064] Concentration and drying: The combined decoctions were concentrated under reduced pressure at 60℃ to a relative density of 1.10-1.20 (measured at 60℃) to obtain an extract. The extract was dried at 60-70℃ until the moisture content was less than 5% to obtain a dry extract.

[0065] Grinding and Packaging: Grind the mixed materials and pass them through a 60-80 mesh sieve to obtain a uniform powder of traditional Chinese medicine additives. Seal the powder in packaging and store it in a cool, dry place.

[0066] Experimental Example 1

[0067] Screening of antioxidant Chinese medicinal herbs

[0068] In order to obtain a drug that can effectively treat liver injury in sea bass, the inventors conducted a large number of screening experiments. Among them, the experimental raw materials screened by the in vitro DPPH antioxidant screening model have achieved good results in treating liver injury in sea bass.

[0069] DPPH is a stable free radical. It exists stably in organic solvents, and its alcoholic solution is purple. It requires low-temperature, light-protected storage. Possessing a single electron, it can accept one electron or a hydrogen ion, and exhibits maximum absorption at 492 nm. In the presence of free radical scavengers, the single electron of DPPH is captured, causing its color to lighten and its absorbance at the maximum absorption wavelength to decrease linearly. This decrease in absorbance indicates increased antioxidant activity, thus evaluating the antioxidant capacity of the test sample. This antioxidant capacity is expressed as an inhibition rate; a higher inhibition rate indicates stronger antioxidant activity.

[0070] Experimental materials and methods:

[0071] The experimental materials are shown in Table 4.

[0072] Table 4. Test Materials

[0073] .

[0074] Test methods

[0075] Extraction of active ingredients from traditional Chinese medicine

[0076] Sample pretreatment

[0077] Place 100g of the test sample in a drying oven and dry at 105℃ to constant weight. Then, pulverize the sample in a pulverizer until it is powdery and pass through a 20-mesh sieve.

[0078] Water extract of the test substance

[0079] The pulverized test substance powder was mixed with water at a ratio of 1:10 and placed into a 50 mL centrifuge tube. The mixture was boiled in a water bath for 2 hours. The extract was then centrifuged at 3000 rpm for 10 minutes. The supernatant was collected, and the resulting solution was the water extract of the test substance. All extracts were calculated at a ratio of 100 mg / mL.

[0080] Antioxidant screening

[0081] Preparation of DPPH solution: Accurately weigh 7.88 mg of DPPH into a beaker and add 80 mL of anhydrous ethanol, mix well. Pour into a 100 mL volumetric flask and dilute to the mark with anhydrous ethanol to prepare a 0.2 mol / mL DPPH solution. Store the solution at 4°C, protected from light. Prepare fresh before use.

[0082] Establishment and stability testing of antioxidant models

[0083] The extracted sample aqueous solutions were prepared at concentrations of 1 mg / mL, 10 mg / mL, and 100 mg / mL using a 10-fold dilution method. Color comparisons were performed to determine the linearity interval. Based on the curve, five gradient concentrations were established for antioxidant assays.

[0084] Antioxidant screening at sample concentrations

[0085] (1) Take 0.2 mL of the sample aqueous solution of the test substance concentration and 3.8 mL of 0.2 mol / mL DPPH solution into a test tube, shake well, and react in the dark for 30 min. Measure its absorbance A at 530 nm using anhydrous ethanol as a blank. a .

[0086] (2) Take 0.2 mL of distilled water and 3.8 mL of 0.2 mol / mL DPPH solution and put them into a test tube. Shake well and react in the dark for 30 min. Use anhydrous ethanol as a blank and measure its absorbance at 530 nm. This is the aqueous control A. b

[0087] R a (%) = (1-A) a / A b )×100%;

[0088] Note: A a Absorbance after adding antioxidant; A b As a control, the IC50 of each sample was calculated based on a 50% scavenging rate to evaluate the antioxidant properties of the samples.

[0089] The results of screening for Chinese medicinal materials with good antioxidant effects are shown in Table 5 below. Figure 2 .

[0090] Table 5. Screening of Antioxidant Chinese Herbal Medicines

[0091] .

[0092] As shown in the table above, gallnut possesses excellent antioxidant capabilities. As a key ingredient in subsequent drug formulation development, other herbs with superior antioxidant capabilities, such as *Gnaphalium affine*, *Centella asiatica*, *Rhizoma Scutellariae*, *Terminalia chebula*, and chlorogenic acid, were selected and prepared into powders using the methods described above. These powders were then added to the feed of California bass at a dosage of 8‰, with chlorogenic acid added at 1‰. The experimental methods are as follows:

[0093] Fatty liver test:

[0094] (1) The experimental feed pellets were approximately 5 mm in diameter, and the experimental animals were juvenile California bass weighing 25-30 g. The fish were temporarily kept in captivity for one week before the experiment began, and the experiment started when the feeding rate reached 3%. Wastewater was discharged daily during the experiment, and the water was changed every two days, with approximately one-third of the water replaced. During the experiment, the water temperature was 25-30℃, ammonia nitrogen concentration <2 mg / L, nitrite concentration <0.01 mg / L, dissolved oxygen content >5.0 mg / L, and pH 7.5-8.0.

[0095] (2) Construction of fatty liver model: The construction method is based on the literature: Li Yu, Zhou Zhigang, He Suxu, et al. Construction of nutritional fatty liver model in zebrafish [J]. Journal of Fisheries of China, 2017, 41(5):10. DOI:10.11964 / jfc.20160810513. In this invention, the method described above is followed. Largemouth bass are continuously fed a high-fat diet with a fat level of 25% (fat level is increased to 25% by adding fat to the conventional diet (fat level is 10-12%)) to induce fatty liver formation in largemouth bass. After the experiment began, each experimental group was fed a high-fat diet for 3 weeks (21 days), fed once each at 08:30, 12:00, and 17:30 daily. The feeding amount was adjusted according to the feeding situation, and the feeding amount of each group of experimental animals was recorded. On the 21st day of the experiment, 3 fish were taken from each tank, and the initial liver condition of each group of experimental animals was recorded. Then, on the 22nd day, they were fed a regular diet with normal fat levels supplemented with the group's medication. After 10 consecutive days of feeding, dissection and sampling were performed, and the liver repair status of each embodiment was recorded. According to Table 1, changes in body surface and liver color were recorded and scored. The results are shown in Table 6.

[0096] Table 6. Liver Injury Score

[0097] ;

[0098] Different letters on the shoulder labels indicate significant differences.

[0099] As shown in Table 6, after 21 days of continuous feeding with a high-fat diet, there were no significant differences in liver disease scores among the initial sampling groups and the model group. P>0.05), obvious fatty liver was observed, mainly manifested as yellowing of the liver. After liver injury, the above-mentioned drugs were added to the feed for 10 consecutive days. The liver injury scores of each of the following examples using Qingguo, Wuluzi, Centella asiatica, Donglingcao, Maochezi, and chlorogenic acid decreased compared with the initial score after injury. Among them, the liver recovery of Qingguo and Wuluzi groups was better, and the liver injury scores decreased significantly compared with other groups. P <0.05, and there was no significant difference compared with the blank control group ( P >0.05), the liver morphology was mainly characterized by a reddish liver with a clear liver edge outline, and there was no significant difference in liver damage scores among the Centella asiatica, Rabdosia rubescens, Terminalia chebula, and chlorogenic acid groups. P >0.05) and significantly higher than that of green fruit, gallnut and blank control group ( P <0.05).

[0100] Further research was conducted using gallnut and green fruit as the core ingredients. Some examples are as follows:

[0101] Example 1

[0102] Sample preparation: Weigh 40 parts of Terminalia chebula, 20 parts of Galla chinensis, 10 parts of Salvia miltiorrhiza, 5 parts of Schisandra chinensis, 10 parts of Glycyrrhiza uralensis, 10 parts of Eucommia ulmoides, and 5 parts of Paeonia lactiflora. The preparation method is the same as in Example 1.

[0103] Example 2

[0104] Sample preparation: Weigh out 50 parts of Terminalia chebula, 15 parts of Galla chinensis, 5 parts of Salvia miltiorrhiza, 15 parts of Schisandra chinensis, 10 parts of Glycyrrhiza uralensis, 10 parts of Eucommia ulmoides, and 5 parts of Paeonia lactiflora. The preparation method is the same as in Example 1.

[0105] Example 3

[0106] Sample preparation: Weigh out 60 parts of green plum, 10 parts of gallnut, 5 parts of salvia miltiorrhiza, 5 parts of schisandra chinensis, 10 parts of licorice, 5 parts of eucommia ulmoides, and 5 parts of white peony root. The preparation method is the same as in Example 1.

[0107] Example 4

[0108] Sample preparation: Weigh 80 parts of green plum, 5 parts of schisandra, 5 parts of licorice, 5 parts of eucommia, and 5 parts of white peony root. The preparation method is the same as in Example 1.

[0109] Example 5

[0110] Sample preparation: Weigh 80 parts of green fruit and 20 parts of gallnut to prepare traditional Chinese medicine additives according to the above preparation method. The preparation method is the same as in Example 1.

[0111] Example 6

[0112] Sample preparation: Weigh 40 parts of Gallnut, 15 parts of Schisandra, 15 parts of Licorice, 15 parts of Eucommia, and 15 parts of White Peony. The preparation method is the same as in Example 1.

[0113] Example 7

[0114] Sample preparation: Weigh 20 parts of Schisandra chinensis, 20 parts of Glycyrrhiza uralensis, 20 parts of Eucommia ulmoides, and 20 parts of Paeonia lactiflora. The preparation method is the same as in Example 1.

[0115] Example 8

[0116] Sample preparation: Weigh 100 portions of gallnuts and prepare them according to the same method as in Example 1.

[0117] Comparative Example 1: Feed the same conventional feed as Examples 1-8, without adding any liver-protecting products.

[0118] In Examples 1-8 above, the total weight of each group of medicinal materials is the same, and the number of portions is to represent the weight ratio.

[0119] Fatty liver test:

[0120] (1) The experimental feed pellets were approximately 5 mm in diameter, and the experimental animals were juvenile California bass weighing 25-30 g. The fish were temporarily kept in captivity for one week before the experiment began, and the experiment started when the feeding rate reached 3%. Wastewater was discharged daily during the experiment, and the water was changed every two days, with approximately one-third of the water replaced. During the experiment, the water temperature was 25-30℃, ammonia nitrogen concentration <2 mg / L, nitrite concentration <0.01 mg / L, dissolved oxygen content >5.0 mg / L, and pH 7.5-8.0.

[0121] (2) Construction of fatty liver model: The construction method is based on the reference: Li Yu, Zhou Zhigang, He Suxu, et al. Construction of nutritional fatty liver model in zebrafish [J]. Journal of Fisheries of China, 2017, 41(5):10. DOI:10.11964 / jfc.20160810513. In this invention, the method described above is followed. Largemouth bass are continuously fed a high-fat diet with a fat level of 25% (fat level is increased to 25% by adding fat to the conventional diet (fat level is 10-12%)) to induce fatty liver formation in largemouth bass. After the experiment began, each experimental group was fed a high-fat diet for 3 weeks (21 days), fed once each at 08:30, 12:00, and 17:30 daily. The feeding amount was adjusted according to the feeding situation, and the feeding amount of each group of experimental animals was recorded. On the 21st day of the experiment, 3 fish were taken from each tank, and the initial liver condition of each group of experimental animals was recorded. Then, on the 22nd day, they were fed a regular diet with normal fat levels supplemented with the group's medication. After 10 consecutive days of feeding, dissection and sampling were performed, and the liver repair status of each embodiment was recorded. Changes in body surface and liver color were recorded according to Table 1, and scores were assigned.

[0122] (3) Liver damage score: The model can generate obvious symptoms that can be observed by the naked eye. The specific symptoms are shown in Table 1. The livers with different degrees of damage are scored, and the results are shown in Table 7.

[0123] Table 7. Liver Disease Scores

[0124] ;

[0125] Different letters on the shoulder labels indicate significant differences.

[0126] Table 6 shows that after feeding California bass with a high-fat diet for 21 consecutive days, all experimental animals in each group developed obvious liver damage symptoms. The main liver damage symptoms were yellow liver, with some animals also exhibiting ascites and jaundice. Milder symptoms were characterized by a yellowish liver color. The liver scores of each embodiment were significantly higher than those of the blank control group. P <0.05), there was no significant difference in symptoms among the various embodiments ( P >0.05); After liver injury, the drugs specified in each example were added and fed continuously for 10 days. The liver injury scores of each example decreased compared with the initial scores after injury. Among them, the livers of Examples 1-5 recovered better, and their liver injury scores were significantly lower than those of Examples 6-8 and Comparative Example 1. P <0.05), there was no significant difference in liver injury scores between Examples 1 and 5 ( P >0.05), Examples 6-8 were also significantly lower than Comparative Example 1 (model group) ( P <0.05), the effect on liver damage repair was relatively poor compared to Examples 1-5.

[0127] The experimental results show that the in vitro antioxidant screening results cannot completely and positively reflect the liver damage repair effect in California bass. However, the experiments concluded that the traditional Chinese medicine additive of this invention can effectively improve the growth performance of California bass, reduce liver fat content, and repair hepatocyte damage, demonstrating significant liver-protective effects.

[0128] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of a traditional Chinese medicine composition in the preparation of drugs or feed for the prevention or treatment of fatty liver in fish, characterized in that, The active ingredients of the traditional Chinese medicine composition are made from the following raw materials in parts by weight: 40-80 parts of green fruit and 10-20 parts of gallnut.

2. The application according to claim 1, characterized in that, In the aforementioned traditional Chinese medicine composition, the weight ratio of green fruit and gallnut is (2~6):

1.

3. The application of a traditional Chinese medicine composition in the preparation of drugs or feed for the prevention or treatment of fatty liver in fish, characterized in that, The active ingredients of the traditional Chinese medicine composition are made from the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Gallnut, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

4. The application according to claim 3, characterized in that, The active ingredients of the traditional Chinese medicine composition are made from the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Gallnut, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5 parts of Paeonia lactiflora.

5. The application according to any one of claims 1 to 4, characterized in that, The weight fraction of green fruit in the traditional Chinese medicine composition is more than 35%.

6. The application according to any one of claims 1 to 4, characterized in that, The weight fraction of green fruit in the traditional Chinese medicine composition is ≥40%.

7. The application according to any one of claims 1 to 4, characterized in that, The dosage form of the traditional Chinese medicine composition is any one of powder, oral liquid, spray, ointment, injection, and capsule.

8. The application according to any one of claims 1 to 4, characterized in that, The fish in question is a perch.

9. The application according to any one of claims 1 to 4, characterized in that, The traditional Chinese medicine composition is a feed additive.

10. The application according to any one of claims 1 to 4, characterized in that, The traditional Chinese medicine composition is obtained by extracting traditional Chinese medicine.

11. The application according to any one of claims 1 to 4, characterized in that, The traditional Chinese medicine composition is an aqueous extract.

12. The application according to any one of claims 1 to 4, characterized in that, The traditional Chinese medicine composition is an extract obtained by decocting powdered medicinal materials with 5 to 15 times the amount of water.

13. The application according to any one of claims 1 to 4, characterized in that, The preparation method of the traditional Chinese medicine composition includes: washing each medicinal material, drying it at 50-60℃, pulverizing it, passing it through a 60-80 mesh sieve, and mixing it evenly; adding 5-15 times the amount of purified water to the mixed medicinal powder, heating it to boiling, and decocting it for 1-3 hours; filtering it to obtain the first decoction; adding 5-10 times the amount of water to the residue again, decocting it for 1-2 hours, and filtering it to obtain the second decoction; combining the two decoctions; concentrating the decoction under reduced pressure at 60℃ to a relative density of 1.10-1.20 to obtain an extract; drying the extract at 60-70℃ until the moisture content is less than 5% to obtain a dry extract; then pulverizing it and passing it through a 60-80 mesh sieve to obtain a uniform traditional Chinese medicine additive powder.

14. A traditional Chinese medicine composition for the prevention or treatment of fatty liver in fish, characterized in that, The active ingredients of the traditional Chinese medicine composition are made from the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Gallnut, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

15. The traditional Chinese medicine composition according to claim 14, characterized in that, The active ingredients of the traditional Chinese medicine composition are made from the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Gallnut, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 5-10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5-10 parts of Paeonia lactiflora.

16. The traditional Chinese medicine composition according to claim 14, characterized in that, The active ingredients of the traditional Chinese medicine composition are made from the following raw materials in parts by weight: 40-80 parts of Terminalia chebula, 10-20 parts of Gallnut, 5-10 parts of Salvia miltiorrhiza, 5-15 parts of Schisandra chinensis, 10 parts of Glycyrrhiza uralensis, 5-10 parts of Eucommia ulmoides, and 5 parts of Paeonia lactiflora.

17. The traditional Chinese medicine composition according to any one of claims 14 to 16, characterized in that, The weight fraction of green fruit in the traditional Chinese medicine composition is more than 35%.

18. The traditional Chinese medicine composition according to any one of claims 14 to 16, characterized in that, The dosage form of the traditional Chinese medicine composition is any one of powder, oral liquid, spray, ointment, injection, and capsule.

19. The traditional Chinese medicine composition according to any one of claims 14 to 16, characterized in that, The fish in question is a perch.

20. The traditional Chinese medicine composition according to any one of claims 14 to 16, characterized in that, The traditional Chinese medicine composition is a drug or feed.

21. The traditional Chinese medicine composition according to any one of claims 14 to 16, characterized in that, The traditional Chinese medicine composition is a feed additive.

22. A method for preparing a traditional Chinese medicine composition according to any one of claims 14 to 21, characterized in that, The process includes the following steps: The traditional Chinese medicine composition is obtained by extracting the traditional Chinese medicine.

23. The preparation method according to claim 22, characterized in that, The traditional Chinese medicine composition is an aqueous extract.

24. The preparation method according to claim 22, characterized in that, The traditional Chinese medicine composition is an extract obtained by pulverizing and decocting in water.

25. The preparation method according to claim 22, characterized in that, The traditional Chinese medicine composition is an extract obtained by decocting powdered medicinal materials with 5 to 15 times the amount of water.

26. The preparation method according to claim 22, characterized in that, The preparation method of the traditional Chinese medicine composition includes: washing each medicinal material, drying it at 50-60℃, pulverizing it, passing it through a 60-80 mesh sieve, and mixing it evenly; adding 5-15 times the amount of purified water to the mixed medicinal powder, heating it to boiling, and decocting it for 1-3 hours; filtering to obtain the first decoction; adding 5-10 times the amount of water to the residue again, decocting it for 1-2 hours, and filtering to obtain the second decoction; combining the two decoctions; concentrating the decoction under reduced pressure at 60℃ to a relative density of 1.10-1.20, and measuring the relative density at 60℃ to obtain an extract; drying the extract at 60-70℃ until the moisture content is less than 5% to obtain a dry extract, then pulverizing it and passing it through a 60-80 mesh sieve to obtain a uniform traditional Chinese medicine additive powder.

27. A fish feed, comprising a basic fish feed, characterized in that, It also includes the traditional Chinese medicine composition according to any one of claims 14 to 21, or the product prepared by any one of claims 22 to 26, which accounts for 1-10‰ of the feed weight.

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