An experimental mouse non-alcoholic fatty liver model feed, and a preparation method and application thereof

Abstract

The present application relates to a kind of experimental mouse non-alcoholic fatty liver model feed and its preparation method and application, belong to animal model feed technical field;The feed includes the following preparation raw materials: casein, fructose, sucrose, cellulose, cholesterol, malt dextrin, L-cystine, mineral mixture, vitamin mixture, choline tartrate, lard, soybean oil, shortening, sodium cholate, traditional Chinese medicine extract and dye agent.The advantage lies in, after using the feed of the present application to feed mouse for a period of time, mouse non-alcoholic fatty liver model can be obtained, the dietary habits of non-alcoholic fatty liver population can be simulated, the modeling process is more gentle, model is more stable, more conducive to the research of disease and drug.The present application can be according to the model index needs, with the feeding mode of automation, customization, to experimental mouse scientific feeding, avoid the situation of not in line with the regulations of feeding time and amount to appear by artificial control, in turn, it is favorable to the integrity and consistency of model, improve the success rate and efficiency of modeling.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of animal model feed, in particular to an experimental mouse non-alcoholic fatty liver model feed and a preparation method and application thereof. BACKGROUND

[0002] Non-alcoholic steatohepatitis (hereinafter also referred to as "NASH") is a histological observation of non-alcoholic patients showing similar to alcoholic hepatitis, and is a progressive liver disease observed with inflammation and fibrosis in liver tissue accompanied by fat deposition.

[0003] The current most commonly used method for constructing a non-alcoholic fatty liver animal model is to use high-fat feed, which has the advantages of simple operation and good repeatability; but also has defects such as long modeling time and high experimental cost. Although the use of high-concentration fat milk gavage or drugs such as carbon tetrachloride intraperitoneal injection can induce non-alcoholic fatty liver by causing acute liver lesions, the modeling time is short, but the success rate of modeling is low, the mortality rate of animals is high, and the pathogenesis, lesion process and histological morphological changes are quite different from human fatty liver. SUMMARY

[0004] The present application aims to solve the problems of long modeling time and low modeling success rate in the related art by providing an experimental mouse non-alcoholic fatty liver model feed.

[0005] The present application also provides a preparation method of the experimental mouse non-alcoholic fatty liver model feed.

[0006] The present application also provides an application of the experimental mouse non-alcoholic fatty liver model feed.

[0007] Specifically, the present application provides an experimental mouse non-alcoholic fatty liver model feed, which comprises the following preparation raw materials in weight percentage:

[0008] Casein: 18% to 22%, fructose 20% to 24%, sucrose: 10% to 15%, cellulose: 3% to 5%, cholesterol: 2% to 4%, malt dextrin: 10% to 12%, L-cystine: 0.25% to 0.4%, mineral mixture: 1% to 2%, vitamin mixture: 1% to 2%, choline tartrate: 0.05% to 0.15%, lard: 2% to 3%, soybean oil: 2% to 4%, shortening: 14% to 16%, sodium cholate: 0.1% to 0.2%, traditional Chinese medicine extract: 1% to 2%, dye: 0.01% to 0.02%;

[0009] The traditional Chinese medicine extract is composed of the following preparation raw materials:

[0010] Radix Astragali, Atractylodes Macrocephala Koidz, Atractylodes Lancea and Codonopsis.

[0011] According to the feed technical solution of the present application, at least the following beneficial effects are achieved:

[0012] Casein: Casein is a phosphorus-calcium binding protein, which belongs to a protein component. It has the function of regulating blood lipids, and can supplement the required calcium elements in the body, thereby promoting the growth of mice.

[0013] Fructose: Fructose is a monosaccharide, which is the most chemically active sugar in sugar. Its metabolism in the human body is faster than that of glucose, and it is easily absorbed by the body; Fructose is almost completely metabolized by the liver, so dietary fructose is pooled into the liver and is mainly metabolized into triglycerides through de novo lipogenesis (DNL).

[0014] Sucrose: Sucrose will hydrolyze to form glucose and fructose, thereby participating in metabolism, being absorbed and converted by the body, and providing energy.

[0015] Cellulose: Promote gastrointestinal peristalsis, keep the intestinal tract unobstructed, and maintain normal food intake.

[0016] Cholesterol: Promote the formation of fatty liver, reduce the time required for modeling.

[0017] Maltodextrin: has thickening effect can to the plasticity of feed, maintain the hardness and shape of the feed; at the same time contains a certain amount of vitamins and trace elements, promote the normal metabolism and metabolism of the body.

[0018] L-cystine: as a feed nutrient enhancer, it is beneficial to animal development, increases body weight and liver and kidney function.

[0019] Mineral mixture: provides the mineral elements required for normal growth and development of animals.

[0020] Vitamin mixture: provides the vitamins required for normal growth and development of animals.

[0021] Choline tartrate: effectively hinders fat metabolism and exacerbates fat accumulation in the liver.

[0022] Soybean oil: contains rich unsaturated fatty acids, enhances lipid metabolism, prevents cardiovascular and cerebrovascular diseases, and has antioxidant and anti-aging effects. It can improve the negative effects of the mouse body caused by eating too much high-fat and high-calorie feed.

[0023] Pig oil: pig oil has high digestibility and absorption rate, and also contains rich vitamin A and vitamin D; thereby increasing the body weight of mice.

[0024] High fat substance (mixture of soybean oil, shortening, lard) means high energy intake, excess energy, mainly in the form of carbohydrates and fats into the liver, relative to the liver will be able to oxidize this energy into CO2. Or lead it out as very low density lipoprotein (VLDLs).

[0025] Sodium cholate: help the body to digest and absorb fat, make fat into tiny droplets, increase the contact area of fat and enzyme, facilitate the decomposition and absorption of fat, can regulate the synthesis of intestinal cholesterol, and at the same time promote the secretion of cholesterol in bile.

[0026] The traditional Chinese medicine extract of the present application is made of Astragalus, Atractylodes, Atractylodes and Codonopsis, wherein the Astragalus extract has obvious effects of reducing blood sugar and improving insulin sensitivity, and can improve the metabolism of substances in the body, thereby promoting the weight gain of mice.

[0027] Codonopsis: Codonopsis extract can increase the content of gastric wall-bound mucus, play a role in resisting gastric mucosal damage, and can increase the longitudinal muscle tension of the gastric fundus, increase the contraction of the gastric body and antral circular muscle.

[0028] Atractylodes: Atractylodes extract can improve appetite, promote digestion and absorption, enhance metabolism, and promote the growth of mice.

[0029] Atractylodes: Atractylodes extract can improve appetite, promote digestion and absorption, enhance metabolism, and promote the growth of mice.

[0030] The traditional Chinese medicine extract of the present application can reduce blood sugar, improve metabolism of substances in the body, and also can promote the excitation of gastrointestinal smooth muscle, thereby promoting gastric juice secretion and gastrointestinal movement, and improving insufficient gastric motility; in addition, it can also improve microcirculation, thereby promoting the weight gain of mice.

[0031] The casein, L-cystine, cellulose, mineral mixture, vitamin mixture and choline tartrate in the feed of the present application ensure that the feed can supply mice with sufficient protein, amino acids, cellulose, minerals, vitamins and choline, which are important for maintaining normal growth and development of mice. The addition of lard, soybean oil, shortening and traditional Chinese medicine extract can improve the non-alcoholic fatty liver rate and palatability of the feed.

[0032] After feeding mice with the feed of the present application for a period of time, a mouse non-alcoholic fatty liver model can be obtained, which can simulate the dietary habits of non-alcoholic fatty liver population, the modeling process is more gentle, the model is more stable, and it is more conducive to the research of diseases and drugs.

[0033] According to some embodiments of the present application, the mineral mixture is composed of the following mass fractions of raw materials:

[0034] Calcium lactate: 20%~30%, iron lactate: 30%~40%, manganese citrate: 15%~20%, zinc citrate: 5%~7%, copper sulfate: 0.1%~0.5%, and aluminum citrate: 10%~15%.

[0035] According to some embodiments of the present application, the vitamin mixture is composed of the following mass fractions of the preparation raw materials:

[0036] Vitamin A: 13%~15%, vitamin D: 2%~3%, vitamin E: 40%~60%, and vitamin K: 20%~30%.

[0037] Controlling the content of vitamin D can increase the IR and inflammatory factor levels of the NAFLD patients to a certain extent, thereby increasing the liver damage and abnormal lipid metabolism phenotype.

[0038] According to some embodiments of the present application, the vitamin mixture is composed of the following mass fractions of the preparation raw materials:

[0039] Vitamin A: 13%~15%, vitamin D: 2%~3%, vitamin E: 50%~60%, and vitamin K: 20%~30%.

[0040] According to some embodiments of the present application, the preparation raw materials of the experimental mouse non-alcoholic fatty liver model further include water.

[0041] According to some embodiments of the present application, the mass fraction of the water is 2%~3%.

[0042] According to some embodiments of the present application, the traditional Chinese medicine extract is composed of the following weight fractions of the preparation raw materials:

[0043] Radix Astragali: 30%~40%, Atractylodes: 5%~10%, Atractylodes lancea: 10%~20%, and Codonopsis pilosula: 30%~40%.

[0044] According to some embodiments of the present application, the preparation method of the traditional Chinese medicine extract includes the following steps:

[0045] After mixing the Radix Astragali, Atractylodes, Atractylodes lancea, and Codonopsis pilosula, the mixture is extracted with an ethanol aqueous solution with a volume fraction of 30%~40%, and then solid-liquid separation is performed, the liquid phase is collected, and concentrated to dryness.

[0046] According to some embodiments of the present application, the extraction temperature is 80°C~90°C.

[0047] According to some embodiments of the present application, the extraction time is 2h~3h.

[0048] According to some embodiments of the present application, the mesh number of the casein is 100 mesh~200 mesh.

[0049] According to some embodiments of the present application, the mesh number of the malt dextrin is 100 mesh to 200 mesh.

[0050] According to some embodiments of the present application, the mesh number of the sucrose is 60 mesh to 100 mesh.

[0051] According to some embodiments of the present application, the dye is a food-grade yellow dye.

[0052] The second aspect of the present application discloses a preparation method of the experimental mouse non-alcoholic fatty liver model feed, comprising the following steps:

[0053] After mixing the preparation raw materials, granulating and drying;

[0054] The temperature of the mixing is 20℃ to 30℃.

[0055] According to some embodiments of the present application, the mixing time is 1h to 2h.

[0056] According to some embodiments of the present application, the drying temperature is 20℃ to 25℃.

[0057] According to some embodiments of the present application, the humidity during the drying process is 20% to 30%.

[0058] According to some embodiments of the present application, the drying time is 24h to 36h.

[0059] The third aspect of the present application discloses the application of the experimental mouse non-alcoholic fatty liver model feed in the modeling of experimental mice.

[0060] According to some embodiments of the present application, the modeling time of the experimental mice is 10 weeks to 12 weeks.

[0061] According to some embodiments of the present application, the feeding density of the experimental mice is 3 to 5 per cage. BRIEF DESCRIPTION OF DRAWINGS

[0062] Figure 1 is the mechanism diagram of the model feed in the embodiments of the present application.

[0063] Figure 2 is the liver diagram after feeding of the control group and the test groups 1 to 5 of the present application.

[0064] Figure 3 is the liver diagram after feeding of the test groups 6 to 10 of the present application. DETAILED DESCRIPTION

[0065] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be described and explained in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and should not be used to limit the present application. Based on the embodiments provided by the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort should fall within the scope of the present application.

[0066] It is obvious that the accompanying drawings in the following description are only some examples or embodiments of the present application, and for those of ordinary skill in the art, the present application can also be applied to other similar scenarios without creative effort based on the drawings. In addition, it can be understood that although the efforts made in the development process can be complex and lengthy, for those of ordinary skill in the art related to the content disclosed in the present application, some design, manufacture or production changes based on the technology disclosed in the present application are only routine technical means and should not be understood as insufficient disclosure of the content disclosed in the present application.

[0067] In the present application, "embodiment" means that the specific features, structures or characteristics described in conjunction with the embodiment can be included in at least one embodiment of the present application. The appearance of this phrase at various places in the specification does not necessarily mean the same embodiment, nor is it an independent or alternative embodiment to other embodiments. It is explicitly and implicitly understood by those of ordinary skill in the art that the embodiments described in the present application can be combined with other embodiments without conflict.

[0068] Unless otherwise defined, the technical terms or scientific terms involved in the present application should be understood as the usual meaning by those of ordinary skill in the art to which the present application belongs. The terms "one", "a", "an", "the" and the like similar words involved in the present application do not represent quantity limitation, but can represent singular or plural. The terms "include", "contain", "have" and any variations thereof involved in the present application are intended to cover non-exclusive inclusion; for example, a process, method, system, product or device including a series of steps or units (units) is not limited to the listed steps or units, but can also include steps or units not listed, or can also include other steps or units inherent to the process, method, product or device. The terms "first", "second", "third" and the like involved in the present application are only to distinguish similar objects, and do not represent a specific order for the objects.

[0069] In the description of the present application, the description of the terms "one embodiment", "some embodiments", "exemplary embodiment", "example", "specific example", or "some examples" means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the exemplary description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0070] Unless otherwise specified, the specific conditions in the embodiments are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are conventional products that can be obtained by commercial purchase, unless otherwise specified.

[0071] The dye used in the embodiments of the present application is a food-grade yellow dye.

[0072] Example 1

[0073] This embodiment is a mouse non-alcoholic fatty liver model feed, which is prepared from the following raw materials in mass fraction:

[0074] Casein (mesh number 150): 19%, fructose: 22%, sucrose (mesh number 60): 11%, cellulose: 4%, cholesterol: 4%, malt dextrin (mesh number 150): 10%, L-cystine: 0.3%, mineral mixture: 2%, vitamin mixture: 2%, choline tartrate: 0.1%, lard: 3%, soybean oil: 4%, shortening: 14%, sodium cholate: 0.1%, traditional Chinese medicine extract: 2%, dye: 0.01%, and water: 2.49%;

[0075] The traditional Chinese medicine extract is prepared from the following raw materials in weight fraction:

[0076] Radix Astragali: 40%, Atractylodes: 10%, Atractylodes lancea: 10%, and Codonopsis pilosula: 40%.

[0077] The mineral mixture in this embodiment is prepared from the following raw materials in weight fraction:

[0078] Calcium lactate: 25%, iron lactate: 38%, manganese citrate: 17%, zinc citrate: 6%, copper sulfate: 0.3%, and aluminum citrate: 13.7%.

[0079] The vitamin mixture in this embodiment is prepared from the following raw materials in weight fraction:

[0080] Vitamin A: 15%, vitamin D: 2%, vitamin E: 53%, and vitamin K: 30%.

[0081] The preparation method of the traditional Chinese medicine extract in the embodiment comprises the following steps:

[0082] The Astragalus membranaceus, Atractylodes macrocephala, Atractylodes lancea and Codonopsis pilosula are ground (the mesh number after grinding is 60 meshes) and mixed, and then extracted by using an ethanol aqueous solution with a volume fraction of 35% (the extraction temperature is 85 ℃, and the extraction time is 3 h), and then subjected to solid-liquid separation, and the liquid phase is collected and concentrated to dryness.

[0083] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in the embodiment comprises the following steps:

[0084] The prepared raw materials are ground (to 200 meshes), mixed, granulated and dried.

[0085] The mixing temperature is 25 ℃, and the mixing time is 1.5 h.

[0086] The drying temperature is 20 ℃, the humidity during the drying process is 20%, and the drying time is 24 h.

[0087] Example 2

[0088] The experimental mouse non-alcoholic fatty liver model feed in the embodiment is prepared from the following mass fractions of prepared raw materials:

[0089] Casein (mesh number: 150 meshes): 20%, fructose: 24%, sucrose (mesh number: 60 meshes): 10%, cellulose: 5%, cholesterol: 2%, maltodextrin (mesh number: 150 meshes): 12%, L-cystine: 0.35%, mineral mixture: 1%, vitamin mixture: 2%, choline tartrate: 0.05%, lard: 2.1%, soybean oil: 3%, shortening: 15%, sodium cholate: 0.2%, traditional Chinese medicine extract: 1%, dye: 0.01%, and water: 2.49%;

[0090] The traditional Chinese medicine extract is prepared from the following weight fractions of prepared raw materials:

[0091] Astragalus membranaceus: 35%, Atractylodes macrocephala: 8%, Atractylodes lancea: 19%, and Codonopsis pilosula: 38%.

[0092] The mineral mixture in the embodiment is prepared from the following weight fractions of prepared raw materials:

[0093] Calcium lactate: 24%, iron lactate: 37%, manganese citrate: 18%, zinc citrate: 7%, copper sulfate: 0.3%, and aluminum citrate: 13.7%.

[0094] The vitamin mixture in the embodiment is prepared from the following weight fractions of prepared raw materials:

[0095] Vitamin A: 15%, vitamin D: 2%, vitamin E: 53%, and vitamin K: 30%.

[0096] The preparation method of the traditional Chinese medicine extract in this embodiment consists of the following steps:

[0097] The Astragalus membranaceus, Atractylodes macrocephala, Atractylodes lancea and Codonopsis pilosula were ground (the mesh number after grinding was 60) and mixed, and then extracted with 35% ethanol aqueous solution (the extraction temperature was 85℃, and the extraction time was 3h) to separate the solid and liquid phases, collect the liquid phase, and concentrate to dryness.

[0098] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in this embodiment was performed according to the method in Example 1.

[0099] Example 3

[0100] The experimental mouse non-alcoholic fatty liver model feed in this embodiment was prepared from the following mass fraction of raw materials:

[0101] Casein (mesh number: 150): 18%, fructose: 23%, sucrose (mesh number: 60): 15%, cellulose: 3%, cholesterol: 3%, maltodextrin (mesh number: 150): 11%, L-cystine: 0.25%, mineral mixture: 1.5%, vitamin mixture: 1.4%, choline tartrate: 0.15%, lard: 2%, soybean oil: 2%, shortening: 16%, sodium cholate: 0.1%, traditional Chinese medicine extract: 1.5%, dye: 0.01%, and water: 2.09%;

[0102] The traditional Chinese medicine extract consisted of the following weight fraction of raw materials:

[0103] Astragalus membranaceus: 35%, Atractylodes macrocephala: 9%, Atractylodes lancea: 17%, and Codonopsis pilosula: 39%.

[0104] The mineral mixture in this embodiment consisted of the following weight fraction of raw materials:

[0105] Calcium lactate: 20%, iron lactate: 40%, manganese citrate: 20%, zinc citrate: 5%, copper sulfate: 0.1%, and aluminum citrate: 14.9%.

[0106] The vitamin mixture in this embodiment consisted of the following weight fraction of raw materials:

[0107] Vitamin A: 14%, vitamin D: 2%, vitamin E: 55%, and vitamin K: 29%.

[0108] The preparation method of the traditional Chinese medicine extract in this embodiment consisted of the following steps:

[0109] Radix Astragali, Rhizoma Atractylodis Macrocephalae, Rhizoma Atractylodis and Radix Codonopsis were ground (the mesh number after grinding was 60 mesh) and mixed, and then extracted with 35% ethanol aqueous solution (the extraction temperature was 85°C, and the extraction time was 3h) to separate the solid and liquid phases, collect the liquid phase, and concentrate to dryness.

[0110] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in this example was performed according to Example 1.

[0111] Example 4

[0112] The experimental mouse non-alcoholic fatty liver model feed in this example was prepared from the following raw materials with the following mass fractions:

[0113] Casein (mesh number: 150 mesh): 21%, fructose: 19%, sucrose (mesh number: 60 mesh): 12%, cellulose: 4.6%, cholesterol: 3%, malt dextrin (mesh number: 150 mesh): 12%, L-cystine: 0.4%, mineral mixture: 1%, vitamin mixture: 1.7%, choline tartrate: 0.1%, lard: 2.85%, soybean oil: 3.1%, shortening: 15%, sodium cholate: 0.15%, traditional Chinese medicine extract: 1.8%, dye: 0.01%, and water: 2.29%;

[0114] The traditional Chinese medicine extract was prepared from the following raw materials with the following weight fractions:

[0115] Radix Astragali: 37%, Rhizoma Atractylodis Macrocephalae: 7%, Rhizoma Atractylodis: 19%, and Radix Codonopsis: 37%.

[0116] The mineral mixture in this example was prepared from the following raw materials with the following weight fractions:

[0117] Calcium lactate: 22%, iron lactate: 37.5%, manganese citrate: 18%, zinc citrate: 7%, copper sulfate: 0.5%, and aluminum citrate: 15%.

[0118] The vitamin mixture in this example was prepared from the following raw materials with the following weight fractions:

[0119] Vitamin A: 13.5%, vitamin D: 2.5%, vitamin E: 55%, and vitamin K: 29%.

[0120] The preparation method of the traditional Chinese medicine extract in this example comprised the following steps:

[0121] Radix Astragali, Rhizoma Atractylodis Macrocephalae, Rhizoma Atractylodis and Radix Codonopsis were ground (the mesh number after grinding was 60 mesh) and mixed, and then extracted with 35% ethanol aqueous solution (the extraction temperature was 85°C, and the extraction time was 3h) to separate the solid and liquid phases, collect the liquid phase, and concentrate to dryness.

[0122] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in this embodiment is carried out with reference to Example 1.

[0123] Example 5

[0124] The experimental mouse non-alcoholic fatty liver model feed in this embodiment is composed of the following preparation raw materials in mass fraction:

[0125] Casein (mesh number 150): 22%, fructose: 22%, sucrose (mesh number 60): 12%, cellulose: 4%, cholesterol: 2%, malt dextrin (mesh number 150): 11%, L-cystine: 0.3%, mineral mixture: 1.8%, vitamin mixture: 1.2%, choline tartrate: 0.1%, lard: 2.3%, soybean oil: 3.2%, shortening: 14.3%, sodium cholate: 0.1%, traditional Chinese medicine extract: 1.4%, dye: 0.01%, and water: 2.29%;

[0126] The traditional Chinese medicine extract is composed of the following preparation raw materials in weight fraction:

[0127] Radix Astragali: 35%, Rhizoma Atractylodis Macrocephalae: 9%, Rhizoma Atractylodis: 19%, and Radix Codonopsis: 37%.

[0128] The mineral mixture in this embodiment is composed of the following preparation raw materials in weight fraction:

[0129] Calcium lactate: 30%, iron lactate: 30%, manganese citrate: 20%, zinc citrate: 6%, copper sulfate: 0.4%, and aluminum citrate: 13.6%.

[0130] The vitamin mixture in this embodiment is composed of the following preparation raw materials in weight fraction:

[0131] Vitamin A: 14%, vitamin D: 2.5%, vitamin E: 56.5%, and vitamin K: 27%.

[0132] The preparation method of the traditional Chinese medicine extract in this embodiment is composed of the following steps:

[0133] Radix Astragali, Rhizoma Atractylodis Macrocephalae, Rhizoma Atractylodis, and Radix Codonopsis are ground (mesh number after grinding: 60 mesh), mixed, extracted with 35% ethanol aqueous solution (extraction temperature: 85°C, extraction time: 3h), and then solid-liquid separated, and the liquid phase is collected and concentrated to dryness.

[0134] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in this embodiment is carried out with reference to Example 1.

[0135] Comparative Example 1

[0136] The experimental mouse non-alcoholic fatty liver model feed in this embodiment is composed of the following preparation raw materials in mass fraction:

[0137] Casein (150 mesh): 19%, fructose: 22%, sucrose (60 mesh): 11%, cellulose: 4%, cholesterol: 4%, malt dextrin (150 mesh): 10%, L-cystine: 0.3%, mineral mixture: 2%, vitamin mixture: 2%, choline tartrate: 0.1%, lard: 3%, soybean oil: 4%, shortening: 14%, sodium cholate: 0.1%, Chinese medicine composition: 2%, dye: 0.01%, water: 2.49%;

[0138] The Chinese medicine composition is composed of the following weight fractions of the preparation raw materials:

[0139] Radix Astragali: 40%, Rhizoma Atractylodis: 10%, Rhizoma Atractylodis: 10%, and Radix Codonopsis: 40%.

[0140] The mineral mixture in this comparative example is composed of the following weight fractions of the preparation raw materials:

[0141] Calcium lactate: 25%, iron lactate: 38%, manganese citrate: 17%, zinc citrate: 6%, copper sulfate: 0.3%, and aluminum citrate: 13.7%.

[0142] The vitamin mixture in this comparative example is composed of the following weight fractions of the preparation raw materials:

[0143] Vitamin A: 15%, vitamin D: 2%, vitamin E: 53%, and vitamin K: 30%.

[0144] The preparation method of the Chinese medicine composition in this comparative example is composed of the following steps:

[0145] Grind (the mesh size after grinding is 60 mesh) and mix Radix Astragali, Rhizoma Atractylodis, Rhizoma Atractylodis, and Radix Codonopsis.

[0146] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in this comparative example is carried out according to Example 1.

[0147] Comparative Example 2

[0148] This comparative example is an experimental mouse non-alcoholic fatty liver model feed composed of the following mass fractions of the preparation raw materials:

[0149] Casein (150 mesh): 19%, fructose: 22%, sucrose (60 mesh): 11%, cellulose: 4%, cholesterol: 4%, malt dextrin (150 mesh): 10%, L-cystine: 0.3%, mineral mixture: 2%, vitamin mixture: 2%, choline tartrate: 0.1%, lard: 3%, soybean oil: 4%, shortening: 14%, sodium cholate: 0.1%, Chinese medicine extract: 2%, dye: 0.01%, water: 2.49%;

[0150] The Chinese medicine extract is composed of the following weight fractions of the preparation raw materials:

[0151] Radix Astragali: 45%, Rhizoma Atractylodis Macrocephalae: 15%, and Radix Codonopsis: 40%.

[0152] The mineral mixture in this comparative example is composed of the following weight fractions of the preparation raw materials:

[0153] Calcium lactate: 25%, iron lactate: 38%, manganese citrate: 17%, zinc citrate: 6%, copper sulfate: 0.3%, and aluminum citrate: 13.7%.

[0154] The vitamin mixture in this comparative example is composed of the following weight fractions of the preparation raw materials:

[0155] Vitamin A: 15%, vitamin D: 2%, vitamin E: 53%, and vitamin K: 30%.

[0156] The preparation method of the Chinese medicine extract in this comparative example is composed of the following steps:

[0157] Radix Astragali, Rhizoma Atractylodis Macrocephalae, and Radix Codonopsis are ground (the mesh size after grinding is 60 mesh), mixed, and then extracted with a 35% volume fraction of an ethanol aqueous solution (the extraction temperature is 85°C, and the extraction time is 3 hours) to separate the solid and liquid phases, collect the liquid phase, and concentrate to dryness.

[0158] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in this comparative example is carried out according to Example 1.

[0159] Comparative Example 3

[0160] The experimental mouse non-alcoholic fatty liver model feed in this comparative example is composed of the following mass fractions of the preparation raw materials:

[0161] Casein (mesh size: 150 mesh): 19%, fructose: 22%, sucrose (mesh size: 60 mesh): 11%, cellulose: 4%, cholesterol: 4%, maltodextrin (mesh size: 150 mesh): 10%, L-cystine: 0.3%, mineral mixture: 2%, vitamin mixture: 2%, choline tartrate: 0.1%, lard: 3%, soybean oil: 4%, shortening: 14%, sodium cholate: 0.1%, Chinese medicine extract: 2%, dye: 0.01%, and water: 2.49%.

[0162] The Chinese medicine extract is composed of the following weight fractions of the preparation raw materials:

[0163] Radix Astragali: 50%, and Radix Codonopsis: 50%.

[0164] The mineral mixture in this comparative example is composed of the following weight fractions of the preparation raw materials:

[0165] Calcium lactate: 25%, iron lactate: 38%, manganese citrate: 17%, zinc citrate: 6%, copper sulfate: 0.3%, and aluminum citrate: 13.7%.

[0166] The vitamin mixture in the present comparative example is composed of the following weight fractions of the preparation raw materials:

[0167] Vitamin A: 15%, vitamin D: 2%, vitamin E: 53%, and vitamin K: 30%.

[0168] The preparation method of the traditional Chinese medicine extract in the present comparative example is composed of the following steps:

[0169] The Astragalus and Codonopsis pilosula are ground (the mesh number after grinding is 60) and mixed, and then extracted with a volume fraction of 35% ethanol aqueous solution (the extraction temperature is 85°C, and the extraction time is 3h) to separate the solid and liquid phases, collect the liquid phase, and concentrate to dryness.

[0170] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in the present comparative example is carried out according to Example 1.

[0171] Comparative Example 4

[0172] The present comparative example is an experimental mouse non-alcoholic fatty liver model feed composed of the following mass fractions of the preparation raw materials:

[0173] Casein (mesh number: 150): 19%, fructose: 22%, sucrose (mesh number: 60): 11%, cellulose: 4%, cholesterol: 4%, maltodextrin (mesh number: 150): 10%, L-cystine: 0.3%, mineral mixture: 2%, vitamin mixture: 2%, choline tartrate: 0.1%, lard: 3%, soybean oil: 4%, shortening: 14%, sodium cholate: 0.1%, traditional Chinese medicine extract (Astragalus extract): 2%, dye: 0.01%, and water: 2.49%;

[0174] The mineral mixture in the present comparative example is composed of the following weight fractions of the preparation raw materials:

[0175] Calcium lactate: 25%, iron lactate: 38%, manganese citrate: 17%, zinc citrate: 6%, copper sulfate: 0.3%, and aluminum citrate: 13.7%.

[0176] The vitamin mixture in the present comparative example is composed of the following weight fractions of the preparation raw materials:

[0177] Vitamin A: 15%, vitamin D: 2%, vitamin E: 53%, and vitamin K: 30%.

[0178] The preparation method of the traditional Chinese medicine extract in the present comparative example is composed of the following steps:

[0179] The Astragalus membranaceus was ground (the mesh number after grinding was 60) and mixed, and then extracted with an ethanol aqueous solution with a volume fraction of 35% (the extraction temperature was 85°C, and the extraction time was 3h) to separate the solid and liquid phases, collect the liquid phase, and concentrate to dryness.

[0180] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in the present comparative example was performed according to Example 1.

[0181] Comparative Example 5

[0182] The present comparative example was an experimental mouse non-alcoholic fatty liver model feed, which was prepared from the following preparation raw materials with the following mass fractions:

[0183] Casein (mesh number: 150): 19%, fructose: 22%, sucrose (mesh number: 60): 11%, cellulose: 4%, cholesterol: 4%, maltodextrin (mesh number: 150): 10%, L-cystine: 0.3%, mineral mixture: 2%, vitamin mixture: 2%, choline tartrate: 0.1%, lard oil: 3%, soybean oil: 4%, shortening: 14%, sodium cholate: 0.1%, dye: 0.01%, and water: 4.49%;

[0184] The mineral mixture in the present comparative example was prepared from the following preparation raw materials with the following weight fractions:

[0185] Calcium lactate: 25%, iron lactate: 38%, manganese citrate: 17%, zinc citrate: 6%, copper sulfate: 0.3%, and aluminum citrate: 13.7%.

[0186] The vitamin mixture in the present comparative example was prepared from the following preparation raw materials with the following weight fractions:

[0187] Vitamin A: 15%, vitamin D: 2%, vitamin E: 53%, and vitamin K: 30%.

[0188] The preparation method of the experimental mouse non-alcoholic fatty liver model feed in the present comparative example was performed according to Example 1.

[0189] Test Example:

[0190] Experimental animals:

[0191] 110 six-week-old C57BL / 6J male mice with a weight of about 20.00g were purchased and selected.

[0192] Group setting:

[0193] Test group 1: fed with the feed in Example 1 of the present application.

[0194] Test group 2: fed with the feed in Example 2 of the present application.

[0195] Test group 3: feeding the feed in Example 3 of the present application.

[0196] Test group 4: feeding the feed in Example 4 of the present application.

[0197] Test group 5: feeding the feed in Example 5 of the present application.

[0198] Test group 6: feeding the feed in Comparative Example 1 of the present application.

[0199] Test group 7: feeding the feed in Comparative Example 2 of the present application.

[0200] Test group 8: feeding the feed in Comparative Example 3 of the present application.

[0201] Test group 9: feeding the feed in Comparative Example 4 of the present application.

[0202] Test group 10: feeding the feed in Comparative Example 5 of the present application.

[0203] Control group: feeding ordinary feed (Jiangsu Cooperation Pharmaceutical Biological Engineering Co., Ltd., item number: XT304).

[0204] The number of mice in each group is 10.

[0205] The parameters during the feeding process of the mice in the test groups of the present application are as follows:

[0206] Each group of mice freely feeds and drinks water every day, the room temperature is maintained at 20-22℃, the humidity is 60%±2%, the length of light is 12h±1h; the feeding time is 11 weeks.

[0207] After 11 weeks, the body weight and the three items of body fat (TC, HDL, LDL) of each mouse are tested; the specific test data are shown in Table 1.

[0208] Table 1: Data of body weight and three items of body fat of mice in the test examples of the present application

[0209]

[0210] At the end of the 11th week, the three items of blood lipid (TC, HDL, LDL) of test groups 1-5 have significant differences with test groups 6-10 and the control group; Figures 2-3 The average is test groups 1-10 (test group 1 corresponds to Figure 2 The average is test group 2 corresponds to Figure 2 The average is test group 3 corresponds to Figure 2 The average is test group 4 corresponds to Figure 2 The average is test group 5 corresponds to Figure 2 The average is test group 6 corresponds to Figure 3 The average is test group 7 corresponds to Figure 3 The average is test group 8 corresponds to Figure 3 The average is test group 9 corresponds toFigure 3 In d, the test group 10 corresponds Figure 3 In e) and the control group Figure 2 In a), the liver after feeding, from the figure: the liver of the test groups 1-5 is enlarged, brownish yellow, the edge is blunt, and the characteristics of fatty liver are obvious, which is more serious. The liver of the test groups 6-10 is red-brown in color, smooth in surface, sharp in edge, and the characteristics of fatty liver are slight; that is, the feed in the embodiments 1-5 of the present application is more likely to promote non-alcoholic fatty liver in mice.

[0211] The present application has the advantages that the feed of the present application has the characteristics of high fat, high fructose, high cholesterol, low methionine, low choline, low VD, etc., fully simulates the dietary mechanism of human non-alcoholic fatty liver, adopts the multiple attack theory, does not limit the supply of feed intake by mice, simulates the overeating diet state of human patients with non-alcoholic fatty liver, simulates various pathogenic factors in the onset process, causes muscle insulin resistance, leads to inhibition of insulin signaling and reduction of insulin-stimulated glucose transport and muscle glycogen synthesis. Since the ingested glucose cannot be correctly stored as muscle glycogen, it is redirected to the liver, where, due to muscle insulin resistance, the ingested glucose is transferred from skeletal muscle glycogen synthesis to the liver for DNL. It combines with compensatory portal hyperinsulinemia, which stimulates SREBP1c, which in turn promotes an increase in the expression of key hepatic enzymes that regulate DNL, leading to increased VLDL production, hypertriglyceridemia, and NAFLD. After feeding mice with the high-fat high-fructose high-cholesterol feed for 10-12 weeks, a mouse model of non-alcoholic fatty liver can be obtained, which can simulate the dietary habits of the non-alcoholic fatty liver population, the modeling process is more gentle, the model is more stable, and it is more conducive to disease and drug research. The present application can scientifically feed experimental mice in an automated and customized manner according to the model index needs, avoid the non-compliance of feeding time and quantity caused by human manipulation, and thus be conducive to the integrity and consistency of the model, improve the success rate and efficiency of modeling.

[0212] The above only describes the preferred embodiments of the present application, and does not limit the implementation and protection scope of the present application. For those skilled in the art, it should be realized that any equivalent replacement and obvious changes made by applying the contents of the present application specification and drawings should be included in the protection scope of the present application.

Claims

1. A feed for an experimental mouse model of non-alcoholic fatty liver disease, characterized in that, The preparation raw materials include the following weight fractions: Casein: 18%~22%, fructose 20%~24%, sucrose: 10%~15%, cellulose: 3%~5%, cholesterol: 2%~4%, maltodextrin: 10%~12%, L-cystine: 0.25%~0.4%, mineral mixture: 1%~2%, vitamin mixture: 1%~2%, choline tartrate: 0.05%~0.15%, lard: 2%~3%, soybean oil: 2%~4%, shortening: 14%~16%, sodium cholate: 0.1%~0.2%, traditional Chinese medicine extract: 1%~2%, dye: 0.01%~0.02%; The traditional Chinese medicine extract is composed of the following weight fractions of preparation raw materials: Radix Astragali: 30%~40%, Rhizoma Atractylodis: 5%~10%, Rhizoma Atractylodis Lancea: 10%~20%, and Radix Codonopsis: 30%~40%; The preparation method of the traditional Chinese medicine extract includes the following steps: After mixing the Radix Astragali, Rhizoma Atractylodis, Rhizoma Atractylodis Lancea, and Radix Codonopsis, the mixture is extracted with an ethanol aqueous solution with a volume fraction of 30%~40%, and then solid-liquid separation is performed, the liquid phase is collected, and concentrated to dryness.

2. The experimental mouse non-alcoholic fatty liver model feed according to claim 1, wherein, The mineral mixture is composed of the following mass fractions of preparation raw materials: Calcium lactate: 20%~30%, iron lactate: 30%~40%, manganese citrate: 15%~20%, zinc citrate: 5%~7%, copper sulfate: 0.1%~0.5%, and aluminum citrate: 10%~15%.

3. The experimental mouse non-alcoholic fatty liver model feed according to claim 1, wherein The vitamin mixture is composed of the following mass fractions of preparation raw materials: Vitamin A: 13%~15%, vitamin D: 2%~3%, vitamin E: 40%~60%, and vitamin K: 20%~30%.

4. The experimental mouse non-alcoholic fatty liver model feed according to claim 1, wherein The preparation raw materials of the experimental mouse non-alcoholic fatty liver model further include water.

5. The experimental mouse non-alcoholic fatty liver disease model feed according to claim 1, wherein The extraction temperature is 80°C~90°C.

6. A method for preparing the experimental mouse non-alcoholic fatty liver model feed according to any one of claims 1 to 5, characterized by, The method includes the following steps: After mixing the preparation raw materials, granulation and drying are performed; The mixing temperature is 20°C~30°C.

7. Use of the experimental mouse non-alcoholic fatty liver model feed according to any one of claims 1 to 5 in modeling of experimental mice.

8. Use according to claim 7, wherein the compound is ###0002### The modeling time of the experimental mice is 10 weeks~12 weeks.

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