阅读说明：本技术 一种高效建立小鼠和大鼠非酒精性脂肪肝模型的饲料配方 (Feed formula for efficiently establishing mouse and rat non-alcoholic fatty liver model ) 是由 吴雨龙 李盛杰 江海涛 霍光明 周峰 徐福飞 华春 于 2020-12-08 设计创作，主要内容包括：本发明公开了一种高效建立小鼠和大鼠非酒精性脂肪肝模型的饲料配方。该配方包括猪油1～15份,蛋黄粉1～15份,胆固醇0.1～3份,胆盐0.1～1份和基础饲料60～90份。动物实验结果表明：该饲料能使ICR小鼠和SD大鼠体重快速增长,在第三周,饲喂配方饲料的小鼠和大鼠体重明显超过饲喂普通饲料的小鼠和大鼠。在第四周,饲喂配方饲料的小鼠和大鼠肝脏出现大量脂滴,基本就能建立小鼠和大鼠非酒精性脂肪肝模型。(The invention discloses a feed formula for efficiently establishing a mouse and rat non-alcoholic fatty liver model. The formula comprises 1-15 parts of lard, 1-15 parts of yolk powder, 0.1-3 parts of cholesterol, 0.1-1 part of bile salt and 60-90 parts of basal feed. The animal experiment results show that: the feed can rapidly increase the body weight of ICR mice and SD rats, and in the third week, the body weight of the mice and rats fed with the formula feed is obviously higher than that of the mice and rats fed with the common feed. In the fourth week, the liver of mice and rats fed with the formula feed shows a large amount of lipid droplets, and then the non-alcoholic fatty liver models of the mice and rats can be basically established.)

1. A feed formula for rapidly establishing mouse and rat non-alcoholic fatty liver models is characterized in that: the formula comprises 1-15 parts of lard, 1-15 parts of yolk powder, 0.1-3 parts of cholesterol, 0.1-1 part of bile salt and 60-90 parts of basal feed.

2. The feed formulation for rapidly modeling non-alcoholic fatty liver disease in mice and rats according to claim 1, wherein: the weight parts of the lard, the yolk powder, the cholesterol, the bile salt and the basic feed are 8-12 parts, 0.5-1.5 parts, 0.1-1 part and 70-80 parts in sequence.

3. The feed formulation for rapidly modeling non-alcoholic fatty liver disease in mice and rats according to claim 1, wherein: the basic feed comprises corn flour, bean pulp, bran, flour, yeast, milk powder, bone meal, rice, fish meal, salt and a compound additive.

4. The feed formulation for rapidly modeling non-alcoholic fatty liver disease in mice and rats according to claim 3, wherein: the composite additive is prepared from the following components in a mass order ratio of 10-30: 1-10: 0.1-3: 0.01-1: 0.5-5 of FeSO₄(7H₂O)、MnSO₄(H₂O)、CuSO₄(5H₂O)、AlCl₃(6H₂O) and ZnSO₄(7H₂O)。

5. The feed formulation for rapidly modeling non-alcoholic fatty liver disease in mice and rats according to claim 3, wherein: 20-30 parts of corn flour, 20-30 parts of soybean meal, 15-20 parts of bran, 15-25 parts of flour, 0.5-1.5 parts of yeast, 0.5-1.5 parts of milk powder, 0.5-1.5 parts of bone meal, 3-8 parts of rice, 3-8 parts of fish meal, 3-8 parts of salt and 0.5-1.5 parts of a composite additive.

6. A method of preparing the feed formulation of claim 1, wherein: the method comprises the steps of proportioning the materials according to a proportion, adding water, mixing, putting into a pellet feed machine to prepare pellet feed with equal particle size, drying in the sun or in the oven, and packaging with a packaging bag for later use.

Technical Field

The invention relates to the field of feed formulas, in particular to a feed formula for efficiently establishing a mouse and rat non-alcoholic fatty liver disease model.

Background

Non-alcoholic fatty liver disease (NAFLD) refers to a clinical pathological syndrome characterized mainly by excessive fat deposition (more than 5% of liver weight) in liver cells caused by alcohol and other definite liver injury factors, and acquired metabolic stress liver injury closely related to insulin resistance and genetic susceptibility. NAFLD has a high incidence in many countries throughout the world. The pathogenesis of NAFLD has not been fully elucidated. Therefore, more and more researchers are beginning to develop drugs for preventing and treating NAFLD, and in the process, NAFLD animal models are often needed, the common animals are mice and rats, and the modeling means is basically to feed formula feed to the mice or rats to form the NAFLD models. At present, the molding formula feed on the market is expensive, about 500 yuan/kg, and causes certain economic pressure for a plurality of scientific research workers.

Disclosure of Invention

Aiming at the technical problems, the invention provides a feed formula for quickly establishing mouse and rat non-alcoholic fatty liver models.

The purpose of the invention can be realized by the following technical scheme:

a feed formula for quickly establishing a mouse and rat non-alcoholic fatty liver model comprises 1-15 parts of lard oil, 1-15 parts of yolk powder, 0.1-3 parts of cholesterol, 0.1-1 part of bile salt and 60-90 parts of basal feed.

The technical scheme of the invention is as follows: the weight parts of the lard, the yolk powder, the cholesterol, the bile salt and the basic feed are 8-12 parts, 0.5-1.5 parts, 0.1-1 part and 70-80 parts in sequence.

The technical scheme of the invention is as follows: the basic feed comprises corn flour, bean pulp, bran, flour, yeast, milk powder, bone meal, rice, fish meal, salt and a compound additive.

The technical scheme of the invention is as follows: the composite additive is prepared from the following components in a mass order ratio of 10-30: 1-10: 0.1-3: 0.01-1: 0.5-5 of FeSO₄(7H₂O)、MnSO₄(H₂O)、Cu SO₄(5H₂O)、AlCl₃(6H₂O) and Zn SO₄(7H₂O)。

The technical scheme of the invention is as follows: 20-30 parts of corn flour, 20-30 parts of soybean meal, 15-20 parts of bran, 15-25 parts of flour, 0.5-1.5 parts of yeast, 0.5-1.5 parts of milk powder, 0.5-1.5 parts of bone meal, 3-8 parts of rice, 3-8 parts of fish meal, 3-8 parts of salt and 0.5-1.5 parts of a composite additive.

A preparation method of the feed formula comprises the steps of proportioning the materials according to a certain proportion, adding water, mixing, putting into a pellet feed machine to prepare pellet feed with equal particle size, drying in the sun or in the oven, and packaging with a packaging bag for later use.

The invention has the beneficial effects that:

the formula feed prepared by the invention has reasonable composition, can efficiently generate mouse or rat NAFLD, and has low price and cost price of about 50 yuan/kg.

Drawings

FIG. 1 shows the effect of normal feed and formula feed on the body weight of male and female mice.

FIG. 2 shows the effect of the common feed and the formula feed on the pathological change of liver tissues of male and female mice, wherein: a: a common feed group; b: group of example 1; c: example 2 group; d: example 3 group. .

FIG. 3 shows the effect of normal and formula feed on the body weight of male rats.

FIG. 4 is a graph showing the effect of conventional feed and formula feed on pathological changes in liver tissue in male rats, wherein: a: a common feed group; b: group of example 1; c: example 2 group; d: example 3 group. .

Detailed Description

The invention is further illustrated by the following examples, without limiting the scope of the invention:

example 1

The feed formula comprises: the parts by weight of the egg yolk powder, the cholesterol, the bile salt and the basal feed are 10kg, 1kg, 0.2kg and 88.8kg in sequence.

Basic feed: 20kg of corn flour, 20kg of soybean meal, 15kg of bran, 15kg of flour, 0.5kg of yeast, 0.5kg of milk powder, 0.5kg of bone meal, 3kg of rice, 3kg of fish meal, 3kg of salt and a compound additive (FeSO)₄(7H₂O)：200g，MnSO₄(H₂O)： 70g，Cu SO₄(5H₂O)：15.6g，AlCl₃(6H₂O)：2.4g，Zn SO₄(7H₂O)：36g)。

Example 2

The feed formula comprises: the weight parts of the lard, the cholesterol, the bile salt and the basic feed are 10kg, 1kg, 0.2kg and 88.8kg in sequence.

Basic feed: 30kg of corn flour, 30kg of soybean meal, 20kg of bran, 25kg of flour, 1.5kg of yeast, 1.5kg of milk powder, 1.5kg of bone meal, 8kg of rice, 8kg of fish meal, 8kg of salt and a compound additive (FeSO)₄(7H₂O)：180g，MnSO₄(H₂O)： 35g，Cu SO₄(5H₂O)：6.8g，AlCl₃(6H₂O)：1.2g，Zn SO₄(7H₂O)：24g)。

Example 3

The feed formula comprises: the weight parts of the lard, the egg yolk powder, the cholesterol, the bile salt and the basic feed are 10kg, 1kg, 0.2kg and 78.8kg in sequence.

Basic feed: 25kg of corn flour, 22kg of soybean meal, 18kg of bran, 20kg of flour, 1kg of yeast, 1kg of milk powder, 1kg of bone meal, 5kg of rice, 5kg of fish meal, 1.816kg of salt and a compound additive (FeSO)₄(7H₂O)：156.4g， MnSO₄(H₂O)：16.2g，Cu SO₄(5H₂O)：2g，AlCl₃(6H₂O)：0.4g，Zn SO₄(7H₂O)：9g)。

Performance test 1:

1. experimental animals: ICR mice, weight 24 + -2 g, 80, sex.

Each group was divided into 8 groups of 10 animals each, including male (normal diet group), male (example 1 group), male (example 2 group), male (example 3 group), female (normal diet group), female (example 1 group), female (formula diet group, example 2 group), and female (example 3 group), and purified water was drunk.

2. Weighing all mice every week, feeding for 4 weeks, collecting blood from orbit after weighing all mice, centrifuging at 3000r/min for 10min, collecting serum, and measuring serum triglyceride and cholesterol content with full-automatic biochemical analyzer; then, mice were sacrificed by cervical dislocation, a small piece of liver at the same position in the liver tissue of each mouse was dissected and immediately fixed in a 10% neutral formalin solution, and tissue sections were prepared by a conventional method. Specifically, dehydrating and transparentizing the liver tissue by ethanol and xylene with different concentrations, embedding the liver tissue in paraffin, trimming the liver tissue, putting the liver tissue in a slicer to cut a slice with the thickness of 4-5 mu m, dewaxing the slice, staining the slice by HE, and observing pathological changes of the liver tissue under a microscope.

3. The experimental results are as follows:

(1) the effect of the general feed and the formula feed on the body weight of the male and female mice is shown in fig. 1.

P < 0.05 compared to the normal feed group.

(2) The effect of the normal feed and the formula feed on the serum triglyceride and cholesterol of the male and female mice, and the results are shown in table 1.

P < 0.05 compared to the normal feed group.

TABLE 1

(3) The effect of the common feed and the formula feed on the pathological change of liver tissues of male and female mice is shown in a figure 2.

The results are summarized as follows: after the male and female mice are fed with the formula feed for 4 weeks, the weight average of the male and female mice is higher than that of the common feed group, and the triglyceride and cholesterol levels in serum are also improved, wherein the group in example 3 has a significant difference compared with the common feed group. The liver tissues in the group in the example 3 can obviously show that the liver index is obviously increased due to the fact that the liver tissues are enlarged in size, yellow, greasy in feeling and blunted at the edges, and the HE stained sections of the liver tissues can show that liver cells are obviously swollen, fat vacuoles are obviously visible in cytoplasm, the fusion of fat drops causes the deviation of cell nuclei, even the cell nuclei disappear, and large oil drops and tiny fat drops which are dispersedly distributed can be seen in the liver tissues and the whole liver is scattered, so that the feed formula in the example 3 can cause the nonalcoholic fatty liver of male and female mice.

Performance detection 2:

1. experimental animals: SD male rats weighing 100 + -2 g, 40.

2. The feed is divided into 4 groups (common feed group and formula feed group, example 1 group, example 2 group and example 3 group), each group contains 10 animals, and purified water is drunk.

3. Weighing all rats every week, feeding for 4 weeks, then weighing all rats, anesthetizing, collecting blood from abdominal aorta, centrifuging at 3000r/min for 10min, collecting serum, and measuring serum triglyceride and cholesterol content with full-automatic biochemical analyzer; the rats were then sacrificed and a small liver piece from the same location in the liver tissue of each rat was dissected and immediately fixed in 10% neutral formalin for tissue sectioning by conventional methods. Specifically, dehydrating and transparentizing the liver tissue by ethanol and xylene with different concentrations, embedding the liver tissue in paraffin, trimming the liver tissue, putting the liver tissue in a slicer to cut a slice with the thickness of 4-5 mu m, dewaxing the slice, staining the slice by HE, and observing pathological changes of the liver tissue under a microscope.

4. The experimental results are as follows:

(1) the effect of the normal feed and the formula on the body weight of male rats is shown in FIG. 3.

P < 0.05 compared to the normal feed group.

(2) The effect of the normal feed and the formula feed on serum triglycerides and cholesterol in male rats and the results are shown in table 2.

TABLE 2

P < 0.05 compared to the normal feed group.

(3) The effect of the conventional feed and the formula feed on the pathological changes of the liver tissues of male rats is shown in FIG. 4.

The results are summarized as follows: after the male rats are fed with the formula feed for 4 weeks, the body weight of the male rats is improved compared with that of a common feed group, and the levels of triglyceride and cholesterol in serum are also improved, wherein the group in the example 3 is obviously different from that of the common feed group. The liver tissues in the group in the example 3 are obviously enlarged in volume, yellow in color, strong in greasy feeling and obviously improved in liver index, and the HE stained section of the liver tissues can show that a large number of vacuoles (lipid droplets) with different sizes are formed in the cytoplasm of liver cells, the boundary is clear, the cell nucleus is extruded to one side, the cytoplasm is loose, the cell nucleus is shrunk, the boundary between the cell and the cell is unclear, and mild inflammation exists, so that the formula feed in the example 3 can cause the nonalcoholic fatty liver of male rats.