Metabolic syndrome high-fat feed composition for rats and preparation process of metabolic syndrome high-fat feed composition

Abstract

The invention belongs to the technical field of experimental animal nutriology, and discloses a high-fat feed composition for mouse metabolic syndrome and a preparation process of the high-fat feed composition. The high-fat feed composition comprises casein VTF, DL-methionine, corn starch, maltodextrin, cane sugar, cellulose, anhydrous milk fat, corn oil, ethoxyquin, a mineral mixture S10001AIN-76A, calcium carbonate, a vitamin mixture 40060-D and cholesterol. The preparation process comprises the following steps: S1, pretreating the raw materials; S2, preparing the materials; S3, mixing the materials; S4, granulating; the high-fat feed provided by the invention can effectively improve the food consumption of mice and ensure the weight growth rate of the mice; and on the basis that the atherosclerosis animal model is stable, the time required for preparing the atherosclerosis mouse is shortened.

Description

Technical Field

[0001] The present invention relates to the technical field of experimental animal nutrition, in particular to a metabolic syndrome high-fat feed composition for mice and a preparation process thereof. Background Art

[0002] With improvements in living standards and irregular diets and rest patterns, the clinical incidence of atherosclerosis (AS) has increased annually. AS includes coronary atherosclerosis, cerebral arteriosclerosis, and mesenteric atherosclerosis. Due to its diverse and complex causes, fundamental research into its pathogenesis has received increasing attention. Therefore, studying animal models of atherosclerosis is fundamental to the clinical discovery and treatment of atherosclerosis.

[0003] High-fat animal models can simulate a high-fat diet and metabolic disorders in humans, exploring the pathogenesis of diseases like hyperlipidemia and obesity and clarifying pathological processes such as abnormal fat metabolism and insulin resistance. In drug development, these models can be used to evaluate the effectiveness and safety of lipid-lowering and weight-loss drugs and identify drug targets. Furthermore, high-fat animal models can be used to study the effects of interventions like diet and exercise on improving metabolic diseases. They can also assist in screening biomarkers, providing a reference for clinical diagnosis and prognosis assessment, and are crucial tools for advancing medical research and clinical practice.

[0004] At present, the common high-fat animal models are SD rats, Wistar rats and APOE rats. - / - However, the establishment time of high-fat animal models using traditional high-fat diet formulas requires 15-18 weeks, which is not suitable for APOE - / - Mice require 10 to 12 weeks, making the modeling process long and the success rate unstable. In addition, long-term feeding of high-fat diets can lead to excessive skin lipid secretion in mice, resulting in seborrheic alopecia and other problems. The laboratory uses a single high-fat diet to model animal obesity, but due to the low palatability of high-fat diets and continuous single feeding, animals can easily develop anorexia and other problems, resulting in low food utilization and unstable weight gain. Summary of the Invention

[0005] The present invention aims to provide a high-fat feed composition for metabolic syndrome in mice and its preparation process, which can effectively increase the food intake of mice and ensure the mice's weight gain rate. Furthermore, while maintaining a stable atherosclerosis animal model, it also shortens the time required to prepare atherosclerosis mice. This solves the problems mentioned in the previous art.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] A high-fat feed composition for metabolic syndrome in mice comprises, by weight, 185-205 parts of casein VTF, 1.5-4.5DL parts of methionine, 40-60 parts of corn starch, 85-115 parts of maltodextrin, 350-370 parts of sucrose, 40-60 parts of cellulose, 190-210 parts of anhydrous milk fat, 8-12 parts of corn oil, 0.04 parts of ethoxyquin, 30-40 parts of mineral mixture S10001AIN-76A, 35-45 parts of calcium carbonate, 8-12 parts of vitamin mixture 40060-D and 1.5 parts of cholesterol.

[0008] Furthermore, the invention comprises, by weight, 195 parts of casein VTF, 3 parts of DL-methionine, 50 parts of corn starch, 100 parts of maltodextrin, 341.46 parts of sucrose, 50 parts of cellulose, 200 parts of anhydrous milk fat, 10 parts of corn oil, 0.04 parts of ethoxyquin, 35 parts of mineral mixture S10001AIN-76A, 39 parts of calcium carbonate, 10 parts of vitamin mixture 40060-D and 1.5 parts of cholesterol.

[0009] The preparation process of the above-mentioned high-fat feed composition for metabolic syndrome in mice comprises the following steps:

[0010] S1. Raw material pretreatment

[0011] Solid raw material grinding: casein VTF, corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A and cholesterol are ground separately;

[0012] Trace ingredient premix: DL-methionine, vitamin mixture 40060-D, and ethoxyquin are trace ingredients. Mix the trace ingredients with corn starch.

[0013] Fat processing: anhydrous milk fat is first melted and then filtered to remove impurities; corn oil is checked for deterioration;

[0014] S2. Ingredients

[0015] According to the content of each raw material of the high-fat feed composition, the required raw materials are weighed by mass, and the weighed raw materials are placed separately for subsequent mixing operations;

[0016] S3, Mixing

[0017] Primary mixing: casein VTF, corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A and cholesterol are put into a mixer for primary mixing;

[0018] Adding premix and fat: Add the premix of DL-methionine, vitamin mixture 40060-D, and ethoxyquin prepared in step S1, and the melted anhydrous milk fat and corn oil into a mixer and continue mixing;

[0019] S4. Granulation

[0020] Conditioning: Steam is introduced into the powdered feed obtained by mixing in step S3 to condition the feed, thereby gelatinizing the corn starch and denaturing the casein VTF, thereby improving the palatability and formability of the high-fat feed composition;

[0021] Compression: The conditioned feed is fed into the pellet mill for compression;

[0022] Cooling: Cool the hot pellet feed obtained by pressing to prevent the feed from getting moldy and deteriorating during storage;

[0023] S5, sterilization, post-processing

[0024] Sterilize the pellet feed prepared in step S4, and screen and package the sterilized pellet feed in sequence;

[0025] S6. Finished product inspection and storage

[0026] Quality inspection: Conduct sensory testing, nutrient analysis, and microbiological testing on finished feed to ensure that the feed quality meets experimental requirements;

[0027] Storage: Store qualified feed in a cool, dry, and ventilated warehouse, avoid direct sunlight and storage with toxic and hazardous substances. Regularly check the quality of inventory feed, follow the first-in-first-out principle, and ensure that the feed is used within the shelf life.

[0028] Furthermore, in S1, the solid raw materials are crushed to 80-100 meshes by using a hammer mill; and the hammer mill is cleaned after each solid raw material is crushed to avoid cross contamination of the solid raw materials.

[0029] Furthermore, the mass ratio of the trace ingredients to the corn starch is 1:10-20, and the trace ingredients and the corn starch are mixed with a mixer for 5-8 minutes to ensure that the trace ingredients are evenly dispersed in the high-fat feed.

[0030] Furthermore, the anhydrous milk fat is melted by heating in a water bath, and the heating temperature is 40-50°C.

[0031] Furthermore, in S2, each raw material is weighed using a high-precision electronic balance or an electronic batching scale.

[0032] Furthermore, in S3, casein VTF, corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A and cholesterol are mixed using a horizontal ribbon mixer, and the speed of the horizontal ribbon mixer is 40 to 60 rpm, and the mixing time is 5 to 8 minutes; when adding the premix and fat, the speed of the horizontal ribbon mixer is adjusted to 60 to 80 rpm, and the mixing time is 10 to 15 minutes to ensure that the fat evenly wraps the raw material particles and the trace components are fully dispersed, and the coefficient of variation of the mixing uniformity is ≤5%.

[0033] Furthermore, in S4, the steam pressure during conditioning is 0.2-0.4 MPa, the conditioning temperature is 75-85° C., the conditioning time is 2-3 minutes, and the moisture content of the feed after conditioning is 16%-18%.

[0034] Furthermore, the diameter of the die hole of the pellet mill is 1 to 2 mm, and the pressure of the pellet mill roller is 0.5 to 0.8 MPa to ensure that the feed passes through the die hole smoothly to form pellets.

[0035] Furthermore, the hot pellet feed is cooled using a counter-current cooler for 8 to 12 minutes. After cooling, the temperature of the pellets drops to room temperature ±5°C, and the moisture content of the feed pellets drops to 12% to 14%.

[0036] Furthermore, in S5, the pellet feed is sterilized using cobalt-60 or electron beam irradiation equipment; the pellet feed is packaged in aluminum foil bags or food-grade plastic sealed bags and divided into packages according to experimental specifications; nitrogen is filled into the bag during packaging to remove air from the bag and delay feed oxidation, and the feed name, formula composition, production date, shelf life, storage conditions, applicable experiment type and sterilization method are clearly marked on the package.

[0037] Furthermore, in S6, sensory testing of finished feed includes testing of color, odor, and particle size; nutrient analysis includes determination of crude protein, crude fat, crude fiber, moisture, ash, cholesterol, and methionine content; microbial testing includes testing of total colony count, E. coli count, and mold count; and storage temperature is 10-25°C, and relative humidity is ≤65%.

[0038] The beneficial effects of the technical solution are:

[0039] 1. The present invention provides a high-fat feed composition for metabolic syndrome in mice, which optimizes the ratio of traditional high-fat feed and the taste of feed, can effectively increase the food intake of mice and ensure the weight gain rate of mice.

[0040] The functions of the components in the high-fat feed composition are as follows:

[0041] Casein VTF: Contains phosphorus-calcium binding protein, which can regulate blood lipids, supplement calcium, relieve muscle soreness and cramps caused by calcium deficiency, improve osteoporosis and growth retardation, and prevent iron deficiency anemia and magnesium deficiency neuritis.

[0042] DL-methionine: A sulfur-containing essential amino acid that protects against oxidative stress and improves animal growth performance.

[0043] Corn starch: provides carbohydrate energy source, increases feed energy density, improves feed color, taste and stability, and promotes animal digestion and absorption of feed.

[0044] Maltodextrin: thickener, enhances feed plasticity, maintains hardness and shape, contains vitamins and trace elements, promotes material metabolism and metabolism.

[0045] Sucrose: A quick source of energy, it promotes animal metabolism, improves the intestinal environment, and increases feed palatability and digestion and absorption rate.

[0046] Cellulose: Promotes intestinal motility, prevents constipation, stimulates gastrointestinal motility, and prevents food residues from staying in the intestines for too long.

[0047] Anhydrous milk fat: a high-energy fat source that provides fatty acids, increases feed energy concentration, improves energy balance, restores body condition, and increases milk production and reproductive performance.

[0048] Corn oil: rich in unsaturated fatty acids and vitamin E, it increases feed energy density, improves taste, promotes the absorption of fat-soluble vitamins, and enhances animals' antioxidant capacity and immune function.

[0049] Ethoxyquin: Antioxidant, protects easily oxidized components in feed, such as vitamins A, D and fat, prevents deterioration, extends the shelf life of feed, and improves feed efficiency.

[0050] Mineral Mix S10001AIN-76A: A purified feed formula that provides a variety of minerals and vitamins to meet the specific nutritional needs of laboratory animals, adjust mineral balance, and support animal growth and health.

[0051] Calcium carbonate: a source of calcium, supplementing calcium to maintain physiological functions such as bone health, nerve conduction and muscle contraction.

[0052] Vitamin Mixture 40060-D: Provides multiple vitamins, such as vitamin D, to promote the absorption and utilization of calcium and phosphorus, maintain bone health, and prevent vitamin deficiency.

[0053] Cholesterol: A component of animal cell membranes, involved in physiological processes such as hormone synthesis, and supports animal growth and development.

[0054] 2. The present invention provides a preparation process for a high-fat feed composition for metabolic syndrome in mice. In the raw material pretreatment stage, the solid raw materials are crushed to 80-100 mesh using a hammer mill, which can significantly increase the specific surface area of ​​the raw materials, promote subsequent mixing uniformity, and avoid physical stimulation of the digestive tract of experimental animals by large particles; the carrier effect of corn starch is used to achieve uniform dispersion of trace ingredients (DL-methionine, vitamin mixture 40060-D and ethoxyquin), solving the industry problem that trace ingredients are easy to agglomerate and difficult to mix; a 40-50°C water bath is used to melt anhydrous milk fat and filter impurities, and the quality of corn oil is simultaneously checked, which not only avoids high temperature destruction of heat-sensitive nutrients, but also ensures the purity of the fat phase and improves the palatability of the feed.

[0055] 3. The present invention provides a preparation process for a high-fat feed composition for metabolic syndrome in mice. During the batching stage, a high-precision electronic balance or electronic batching scale is used to control the mass error of each component, meet the strict requirements of the metabolic syndrome model for the accuracy of feed ingredients, and ensure the repeatability of experimental results.

[0056] 4. The present invention provides a preparation process for a high-fat feed composition for metabolic syndrome in mice. In the mixing stage, a horizontal ribbon mixer is used to mix the main raw materials at 40 to 60 rpm for 5 to 8 minutes, and macroscopic uniformity is achieved through mechanical shearing and convection. In the stage of adding premix and fat, the speed of the horizontal ribbon mixer is increased to 60 to 80 rpm and the mixing time is extended to 10 to 15 minutes. The lubricating effect of fat is used to achieve microscopic uniform coating of the raw material particles. The coefficient of variation of mixing uniformity is ≤5%, which significantly improves the bioavailability of nutrients.

[0057] 5. The present invention provides a preparation process for a high-fat feed composition for metabolic syndrome in mice. During pelleting, the composition is tempered at 75-85°C for 2-3 minutes using 0.2-0.4 MPa steam to moderately gelatinize the corn starch and denature the casein VTF, while simultaneously adjusting the feed moisture content to 16%-18%, significantly improving the feed palatability and pellet formability. A die hole diameter of 1-2 mm and a roller pressure of 0.5-0.8 MPa are combined to ensure pellet hardness while reducing energy consumption and increasing the pellet durability index. Furthermore, the pellet temperature is reduced to room temperature ±5°C through 8-12 minutes of countercurrent cooling, while the moisture content is simultaneously reduced to 12%-14%, effectively preventing pellet cracking and microbial growth.

[0058] 6. The present invention provides a preparation process for a high-fat feed composition for metabolic syndrome in mice. In the sterilization and post-processing stages, cobalt-60 or electron beam irradiation is used to achieve residue-free sterilization, with a sterilization efficiency of 99.99%, significantly extending the shelf life of the feed.

[0059] 7. The present invention provides a preparation process for a high-fat feed composition for metabolic syndrome in mice. During finished product inspection and storage, a detection system covering sensory (color, odor, and particle size), nutritional (six indicators such as crude protein and crude fat), and microbial (three indicators such as total colony count) testing is established to ensure that the feed meets the nutritional standards of experimental animals. The feed is packaged in nitrogen-filled aluminum foil bags and key information is marked with smart labels to achieve traceability of feed quality. Storage conditions are specified to be a constant temperature of 10-25°C and a relative humidity of ≤65%. A real-time temperature and humidity monitoring system is used to ensure feed stability, and a first-in-first-out management system is used to improve nutrient retention. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 This is a comparison chart of the high-fat feed prepared in Example 3 of the present invention and the existing feed;

[0061] In the figure, (a) is conventional maintenance feed, (b) is traditional high-fat feed, and (c) is improved high-fat feed.

[0062] Figure 2 These are representative images of Oil-red staining of aortic plaques in each group of mice in the validation examples of the present invention.

[0063] In the figure, CON is the conventional maintenance feed, 88137 is the traditional high-fat feed, and 7019 is the improved high-fat feed. DETAILED DESCRIPTION

[0064] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments:

[0065] Example 1: A high-fat feed composition for metabolic syndrome in mice, comprising, by weight, 185-205 parts of casein VTF, 1.5-4.5DL parts of methionine, 40-60 parts of corn starch, 85-115 parts of maltodextrin, 350-370 parts of sucrose, 40-60 parts of cellulose, 190-210 parts of anhydrous milk fat, 8-12 parts of corn oil, 0.04 parts of ethoxyquin, 30-40 parts of mineral mixture S10001AIN-76A, 35-45 parts of calcium carbonate, 8-12 parts of vitamin mixture 40060-D and 1.5 parts of cholesterol.

[0066] Example 2: A high-fat feed composition for metabolic syndrome in mice, comprising, by weight, 195 parts of casein VTF, 3 parts of DL-methionine, 50 parts of corn starch, 100 parts of maltodextrin, 341.46 parts of sucrose, 50 parts of cellulose, 200 parts of anhydrous milk fat, 10 parts of corn oil, 0.04 parts of ethoxyquin, 35 parts of mineral mixture S10001AIN-76A, 39 parts of calcium carbonate, 10 parts of vitamin mixture 40060-D, and 1.5 parts of cholesterol.

[0067] Example 3: A process for preparing a high-fat feed composition for metabolic syndrome in mice, comprising the following steps:

[0068] S1. Raw material pretreatment

[0069] Solid raw material crushing: casein VTF, corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A, and cholesterol were respectively put into a hammer mill and crushed to 80-100 mesh; fine particle size can increase the surface area of ​​the raw materials, facilitate subsequent mixing, and facilitate digestion and absorption by mice. During the crushing process, care should be taken to avoid cross-contamination of raw materials, and the equipment should be cleaned after each raw material is crushed.

[0070] Premixing trace ingredients: DL-methionine, vitamin mixture 40060-D, and ethoxyquin are trace ingredients. Premix them with 15 times their weight of corn starch in a small mixer for 5-8 minutes to create a premix with uniform concentration. Premixing ensures that the trace ingredients are evenly dispersed in the feed, preventing localized high or low concentrations from affecting mouse health and experimental results.

[0071] Fat treatment: Anhydrous milk fat needs to be heated in a water bath to 40-50°C to melt, and filtered to remove any impurities. Corn oil can be used directly after inspection to ensure it is not deteriorated.

[0072] S2. Ingredients

[0073] According to the feed formula requirements, use a high-precision electronic balance or electronic batching scale to weigh each raw material. Due to the large variety of raw materials and the small amount of some added, the formula must be carefully checked during weighing to avoid mistakes. The weighed raw materials should be placed in order for subsequent mixing operations.

[0074] S3, Mixing

[0075] Primary mixing: corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A, and cholesterol are sequentially put into a horizontal ribbon mixer, set the speed to 40-60 rpm, and mix for 5-8 minutes to make these basic raw materials preliminarily mixed;

[0076] Add premix and fat: Add the prepared DL-methionine, vitamin mixture 40060-D, and ethoxyquin premix, as well as the melted anhydrous milk fat and corn oil, to the mixer and continue mixing for 10 to 15 minutes. At this time, increase the mixer speed to 60 to 80 rpm to ensure that the fat evenly coats the raw material particles, the trace ingredients are fully dispersed, and the coefficient of variation of the mixing uniformity is controlled to ≤5%;

[0077] S4. Granulation

[0078] Conditioning: Steam is introduced into the mixed powdered feed for conditioning. The steam pressure is controlled at 0.2-0.4 MPa and the conditioning temperature is maintained at 75-85°C for 2-3 minutes to make the moisture content of the feed reach 16%-18%. Steam conditioning can gelatinize corn starch and denature casein, thereby improving the palatability and formability of the feed.

[0079] Pressing: Use a pellet mill die with a die hole diameter of 1-2 mm, and feed the tempered feed into the pellet mill for pressing. During the pressing process, adjust the roller pressure of the pellet mill to 0.5-0.8 MPa to ensure that the feed passes through the die hole smoothly to form pellets with moderate hardness, avoiding pellets that are too hard or too soft.

[0080] Cooling: Use a counter-current cooler to cool the hot pellet feed just pressed. The cooling time is 8 to 12 minutes, so that the pellet temperature drops to close to room temperature (±5°C) and the moisture content drops to 12% to 14%, to prevent the feed from getting moldy and deteriorating during storage.

[0081] S5, sterilization, post-processing

[0082] After the feed is pelletized, cooled, and screened, it is sterilized. The feed is placed in a cobalt-60 or electron beam irradiation device, with the irradiation dose controlled between 15 and 25 kGy. Irradiation sterilization has strong penetrating power and can effectively kill bacteria, mold, parasite eggs, and other microorganisms in the feed, while having minimal damage to the feed's nutritional components. However, care should be taken to avoid oxidation of the feed due to excessive irradiation.

[0083] Secondary screening (if necessary): After irradiation or microwave sterilization, feed pellets generally do not undergo significant changes. If high-pressure steam sterilization is used, some pellets may become soft or sticky due to the heat, and they need to be screened again through a vibrating screen to remove unqualified pellets and ensure uniform product particle size.

[0084] Packaging: Use aluminum foil bags or food-grade plastic sealed bags and package according to common experimental specifications (such as 500 g / bag, 1 kg / bag); fill the bag with nitrogen to remove air and delay feed oxidation; clearly mark the feed name, formula composition, production date, shelf life, storage conditions, applicable experiment type, and sterilization method on the package;

[0085] S6. Finished product inspection and storage

[0086] Quality inspection: sensory testing (observing color, odor, and particle size) of finished feed, nutrient analysis (determining crude protein, crude fat, crude fiber, moisture, ash, cholesterol, methionine content, etc., with a focus on the impact of sterilization on heat-sensitive nutrients), and microbial testing (total colony count, E. coli count, mold count, ensuring that microbial indicators meet standards after sterilization) are performed on finished feed to ensure that feed quality meets experimental requirements;

[0087] Storage: Store qualified feed in a cool, dry, ventilated warehouse with a temperature controlled at 10-25°C and a relative humidity ≤65%; avoid direct sunlight and storage with toxic and hazardous substances. Check the quality of the inventory feed regularly and follow the first-in-first-out principle to ensure that the feed is used within the shelf life.

[0088] The improved high-fat feed was prepared, which was compared with the existing conventional maintenance feed and traditional high-fat feed. Figure 1 As shown in the figure, the improved high-fat feed prepared in this embodiment has a smooth surface and good particle size.

[0089] Verification Example: Animal experiments were performed to verify the performance of the improved high-fat feed prepared in Example 3.

[0090] 1.1 Experimental Animals

[0091] The experimental animals used in this verification case were SPF grade APOE - / - Mice weighing 25 ± 5 g were provided by Beijing Weitonglihua Company under the Laboratory Animal Use Permit No. SYXK(Min)2014-0001. Animal experiments were conducted in accordance with the "Guidelines for the Treatment of Laboratory Animals" (2006) issued by the Ministry of Science and Technology of China and the "Guide for the Care and Use of Laboratory Animals" issued by the National Institutes of Health. Mice were housed in a SPF-grade laboratory at the Animal Experimental Center of Fujian University of Traditional Chinese Medicine. This experiment was approved by the Animal Care and Use Committee of Fujian University of Traditional Chinese Medicine and strictly adhered to the Laboratory Animal Ethics and the regulations of the Fujian University of Traditional Chinese Medicine Animal Care Association (Ethics Review Number: 2W2023099). The animals had free access to food and water. The room was well-ventilated and lighted, with a room temperature of (25 ± 1)°C, a relative humidity of approximately 60%, and a 12-h light / dark cycle. Animals were acclimated for seven days prior to the experiment and maintained in a quiet and undisturbed environment.

[0092] 1.2 Animal grouping

[0093] Six SPF male APOE mice aged 6-8 weeks and weighing 23 g ± 3 g were collected. - / - Mice were randomly divided into a traditional high-fat diet group and a modified high-fat diet group, and given different diets. The experiment was terminated after 20 weeks of high-fat feeding.

[0094] 1.3 Blood pressure measurement

[0095] In this study, the animal's blood pressure was monitored using a noninvasive tail artery blood pressure measurement method. The instrument used was a Kent noninvasive blood pressure meter from the United States. This device uses volume pressure transducer (VPR) technology to measure systolic blood pressure (SBP) and diastolic blood pressure (DBP) in the tail artery of small animals and calculate the mean blood pressure (MAP). Before using this device to measure blood pressure, the animal must be calm and placed in a restraint. An occlusion cuff is placed approximately 1 cm from the base of the tail, followed by the volume pressure transducer (VPR) until the transducer can no longer be pushed toward the base of the tail. The restraint is placed on a heating plate, and the tail is heated using the appropriate heating setting, depending on the room temperature. Before starting the instrument, the animal is covered with a black towel and the experimental environment is kept quiet. After resting in darkness for 10 minutes, blood pressure measurement begins. The occlusion cuff and VPR transducer are inflated and deflated 15 times. The computer software is set up, and the 15 measurements are averaged.

[0096] 1.4 Oil red staining of the abdominal aorta

[0097] like Figure 2 As shown in the figure, the segment from the mouse aortic arch to the renal artery branch of the abdominal aorta was removed, the aorta was cut longitudinally with scissors, the aorta was laid flat, and freshly prepared Oil Red O dye was added to cover it. After staining for 15 minutes, the dye was discarded and the tissue was rinsed with 60% isopropyl alcohol until the normal tissue turned milky white. It was rinsed again with normal saline and observed and photographed. Figure 2 It can be seen that no obvious traces of oil red staining were observed in the aortic tissue of mice in the control group (con). Compared with the control group, the oil red staining range was significantly increased in the traditional high-fat diet (88137) and modified high-fat diet (7019) groups. At 12 weeks, compared with the control group, the two high-fat diets significantly increased the formation of aortic plaques, and both were statistically significant (P < 0.05). At 16 weeks, there was no significant change in the aortic plaques induced by the traditional high-fat diet, but the modified high-fat diet showed a significant difference compared with the control group (P < 0.01). At 20 weeks, the modified high-fat diet showed a significant difference compared with the control group (P < 0.01) and a significant difference compared with the traditional high-fat diet (P < 0.05). The experimental results suggest that the modified high-fat diet is superior to the traditional high-fat diet in terms of the speed and range of aortic plaque formation.

[0098] 1.5 Upper serum test blood glucose, total cholesterol, triglyceride, high-density lipoprotein, low-density lipoprotein content measurement

[0099] After blood was collected from the mouse eyeball, the whole blood was allowed to stand at room temperature for 4 h and then separated into serum using a 4°C centrifuge. The upper serum was taken to detect blood glucose, total cholesterol, triglyceride, high-density lipoprotein, and low-density lipoprotein levels.

[0100] 2. Experimental Results

[0101] 2.1 Effects of different high-fat diets on mouse body weight

[0102] The effects of different high-fat diets on mouse body weight are shown in Table 1. As shown in Table 1, over time (weeks, 0-20 weeks), the body weight (g) of the control group (CON), the traditional high-fat diet group (88137), and the modified high-fat diet group (7019) all showed an upward trend. Compared with the control group, the traditional high-fat diet group and the modified high-fat diet group both had significantly higher body weights than the control group after 1 week. Compared with the traditional high-fat diet, the modified high-fat diet group had significantly higher values ​​than the traditional high-fat diet group after 3 weeks, with statistical significance (P < 0.05). After 9 weeks, the modified high-fat diet group had significantly higher values ​​than the traditional high-fat diet group, with statistical significance (P < 0.01). These experimental results indicate that the modified high-fat diet has a significantly better effect on body weight than the traditional high-fat diet.

[0103] Table 1 Body weight of mice in each group

[0104]

[0105]

[0106] Note: ## indicates P < 0.01 compared with the control group, * indicates P < 0.05 compared with the traditional feed, and ** indicates P < 0.01 compared with the traditional feed.

[0107] 2.2 Effects of different high-fat diets on blood pressure in mice

[0108] The effects of different high-fat diets on blood pressure in mice are shown in Tables 2 to 4. Table 2 shows that the initial systolic blood pressure of mice in the control group was 106.00±3.03 mmHg, that of mice in the traditional high-fat diet group was 104.33±6.12 mmHg, and that of mice in the modified high-fat diet group was 102.50±4.68 mmHg. Significant differences in systolic blood pressure were observed after intervention: SBP in the traditional high-fat diet group increased significantly from week 4 (109.50±4.04 mmHg) to week 20, reaching 133.83±2.71 mmHg. In contrast, systolic blood pressure in the modified high-fat diet group was significantly higher than that in both the control and traditional diet groups (113.50±2.67 mmHg, ##) from week 2, and the rate of increase was even faster, reaching 145.83±5.23 mmHg at week 20. Blood pressure in the control group remained stable, ranging from 102.50 to 114.00 mmHg, with minimal fluctuations.

[0109] As shown in Table 3, the initial diastolic blood pressure (DBP) of mice in the control group was 72.83±5.91 mmHg, that of mice in the traditional high-fat diet group was 70.50±4.51 mmHg, and that of mice in the modified high-fat diet group was 73.83±4.26 mmHg. Significant differences were observed in the changes in DBP after intervention: the traditional high-fat diet group showed a significant increase in DBP from week 4 (82.33±2.94 mmHg, ), reaching 96.00±4.47 mmHg by week 20. In contrast, the modified high-fat diet group showed a significantly higher DBP than both the control and traditional diet groups from week 2 (86.50±5.01 mmHg, ≥5). The rate of increase was even faster, reaching 122.00±3.41 mmHg by week 20. In the control group, DBP showed a slow upward trend from week 0 to week 20 (72.67-81.83 mmHg), with minimal fluctuations and no significant increase.

[0110] As shown in Table 4, the initial mean arterial pressure (MAP) of mice in the control group was 83.86±4.86 mmHg, that of mice in the traditional high-fat diet group was 81.90±3.60 mmHg, and that of mice in the modified high-fat diet group was 83.38±2.83 mmHg. Significant differences in blood pressure changes were observed after intervention: BP in the traditional high-fat diet group increased significantly from week 4 (91.39±1.95 mmHg, ) to 108.61±3.06 mmHg at week 20. In contrast, BP in the modified high-fat diet group was significantly higher than that in both the control and traditional diet groups (95.57±2.60 mmHg, ) at week 2, and the rate of increase was even faster, reaching a high of 129.94±3.76 mmHg at week 20. BP in the control group showed a gradual upward trend (83.86 at week 0 → 92.56 mmHg at week 20), with minimal fluctuations and no significant increase.

[0111] The experimental results show that a high-fat diet can induce increased blood pressure in mice, among which the pressor effect of the modified high-fat diet is more prominent, with an earlier onset and a greater increase, and the degree of damage to blood pressure homeostasis is significantly stronger than that of the traditional high-fat diet.

[0112] Table 2 Systolic blood pressure levels of mice in each group

[0113]

[0114] Note: ## indicates P < 0.01 compared with the control group, * indicates P < 0.05 compared with the traditional feed, and ** indicates P < 0.01 compared with the traditional feed.

[0115] Table 3 Diastolic blood pressure levels of mice in each group

[0116]

[0117]

[0118] Note: # indicates P < 0.05 compared with the control group, ## indicates P < 0.01 compared with the control group, * indicates P < 0.05 compared with the traditional feed, ** indicates P < 0.01 compared with the traditional feed.

[0119] Table 4 Mean arterial pressure of mice in each group

[0120]

[0121] Note: # indicates P < 0.05 compared with the control group, ## indicates P < 0.01 compared with the control group, * indicates P < 0.05 compared with the traditional feed, ** indicates P < 0.01 compared with the traditional feed.

[0122] 2.3 Effects of different high-fat diets on blood lipids and blood glucose in mice

[0123] The effects of different high-fat diets on blood lipids and blood glucose in mice are shown in Tables 5 to 11. As shown in Table 5, the fasting blood glucose level in the control group was 5.13±0.88 mmol / L, which was within the normal range with minimal fluctuations; the fasting blood glucose level in the traditional high-fat diet group increased to 8.73±2.16 mmol / L (P<0.01), suggesting that the traditional high-fat diet can induce elevated blood glucose levels; the modified high-fat diet group significantly increased to 18.44±1.12 mmol / L (P<0.01 compared to both the control and traditional high-fat diet groups, representing a 259.4% increase compared to the control group and a 111.2% increase compared to the traditional group).

[0124] As shown in Table 6, the triglyceride level in the control group was maintained at 0.62-0.92 mmol / L during the 12-20 weeks, and the level was stable. The triglyceride level in the traditional high-fat diet group increased to 2.67±0.56 mmol / L at 12 weeks (P<0.01 compared with the control group, and increased by 190.2% compared with the control group), 3.06±0.40 mmol / L at 16 weeks (P<0.01 compared with the control group), and slightly decreased to 2.02±1.18 mmol / L at 20 weeks (P<0.01 compared with the control group). P < 0.01), but it was still significantly higher than the control group; the triglyceride level in the modified high-fat diet group reached 5.54 ± 0.15 mmol / L at 12 weeks (P < 0.01 compared with the control group, an increase of 502.2% compared with the control group, and an increase of 107.5% compared with the traditional group), and continued to rise to 5.66-5.88 mmol / L at 16-20 weeks (P < 0.01 compared with the control group), which was always the highest among the three groups. The experimental results show that the effect of the modified high-fat diet on TG increase is longer-lasting and stronger.

[0125] As shown in Table 7, the total cholesterol level in the control group remained stable at 3.13-3.51 mmol / L during the 12-20 weeks. In the traditional high-fat diet group, the total cholesterol level rose sharply to 19.57±3.22 mmol / L during the 12-week period (P<0.01 compared with the control group, representing a 457.5% increase compared with the control group), and remained at 20.21-17.69 mmol / L during the 16-20-week period (P<0.01 compared with the control group). In the modified high-fat diet group, the total cholesterol level was 21.57±0. The serum lipid level in the serum of the rats was 43mmol / L (P<0.01 compared with the control group, increased by 514.5% compared with the control group, and increased by 10.2% compared with the traditional group), and significantly increased to 26.15±1.39mmol / L at 16 weeks (P<0.01 compared with the control group and the traditional high-fat diet, increased by 664.6% compared with the control group, and increased by 29.4% compared with the traditional group), and maintained at 25.67±1.62mmol / L at 20 weeks (P<0.01 compared with the control group and the traditional high-fat diet).

[0126] As shown in Table 8, the high-density lipoprotein cholesterol in the control group remained stable at 2.11-2.22 mmol / L during the 12-20 weeks; the high-density lipoprotein cholesterol in the traditional high-fat diet group rose slightly to 2.36±0.21 mmol / L during the 12-week period, and dropped to 1.99-2.04 mmol / L during the 16-20 weeks period, with no significant difference from the control group; the high-density lipoprotein cholesterol in the modified high-fat diet group was 2.59±0.39 mmol / L during the 12-week period, and dropped significantly to 1.92-1.51 mmol / L during the 16-20 weeks period (P<0.01 compared with both the control group and the traditional high-fat diet at 20 weeks, a decrease of 28.4% compared with the control group and a decrease of 25.0% compared with the traditional group). The experimental results show that the modified high-fat diet may specifically inhibit the synthesis of high-density lipoprotein cholesterol and aggravate lipid metabolism disorders.

[0127] As shown in Table 9, the low-density lipoprotein cholesterol in the control group was maintained at 0.18-0.22mmol / L during the 12-20 weeks, which was at an extremely low level. The low-density lipoprotein cholesterol in the traditional high-fat diet group increased sharply to 13.01±1.49mmol / L during the 12-week period (P<0.01 compared with the control group, an increase of 722.8% compared with the control group), and was maintained at 13.43~8.62mmol / L during the 16-20 weeks (P<0.01 compared with the control group). The low-density lipoprotein cholesterol in the modified high-fat diet group was 13.2 at week 12. 7±0.80mmol / L (P<0.01 compared with the control group, an increase of 730.6% compared with the control group, and an increase of 2.0% compared with the traditional group), and continued to rise to 14.15-14.28mmol / L in 16-20 weeks (P<0.01 compared with the control group and the traditional high-fat diet, and an increase of 65.7% compared with the traditional group in 20 weeks). The experimental results show that the improved high-fat diet can strongly promote the increase of low-density lipoprotein cholesterol that causes atherosclerosis, and the effect continues to increase.

[0128] As shown in Table 10, the TG / HDL-C ratio of the control group was maintained at 0.29-0.42 during the 12-20 weeks, indicating metabolic health; that of the traditional high-fat diet group increased to 1.14±0.26 during the 12-week period (P<0.01 compared with the control group, and an increase of 171.4% compared with the control group), and to 1.57-1.02 during the 16-20 weeks (P<0.01 compared with the control group); that of the modified high-fat diet group increased to 2.18±0.31 during the 12-week period (P<0.01 compared with both the control group and the traditional high-fat diet, an increase of 419.0% compared with the control group, and an increase of 91.2% compared with the traditional group), and soared to 2.85-3.90 during the 16-20 weeks (P<0.01 compared with both the control group and the traditional high-fat diet, and an increase of 282.4% compared with the traditional group at 20 weeks). The experimental results show that the lipid metabolism imbalance caused by the modified high-fat diet is more serious.

[0129] As shown in Table 11, the TyG index of the control group remained at 28.42-33.23 during the 12-20 weeks, with no signs of insulin resistance; the TyG index of the traditional high-fat diet group rose sharply to 287.05±77.51 during the 12-week period (P<0.01 compared with the control group, an increase of 764.1% compared with the control group), and reached 377.39-157.80 during the 16-20 weeks (P<0.01 compared with the control group); the TyG index of the modified high-fat diet group rose to 516.35±45.53 during the 12-week period (P<0.01 compared with both the control group and the traditional high-fat diet, an increase of 1454.0% compared with the control group and an increase of 80.0% compared with the traditional group), and soared to 109.81±11.00 during the 16-20 weeks.

[0130] 865.00-974.01 (P < 0.01 compared with the control group and traditional high-fat diet, and increased by 517.2% compared with the traditional group at 20 weeks). The experimental results showed that the improved high-fat diet can strongly induce insulin resistance, and the degree is far greater than that of the traditional high-fat diet.

[0131] Table 5 Fasting blood glucose levels of mice in each group

[0132]

[0133] Note: ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0134] Table 6 Serum triglyceride levels of mice in each group

[0135]

[0136]

[0137] Note: ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0138] Table 7 Serum total cholesterol levels of mice in each group

[0139]

[0140] Note: ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0141] Table 8 Serum high-density lipoprotein cholesterol levels in each group of mice

[0142]

[0143] Note: ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0144] Table 9 Serum low-density lipoprotein cholesterol levels in each group of mice

[0145]

[0146] Note: ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0147] Table 10 Serum TG / HDL-C of mice in each group

[0148]

[0149] Note: ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0150] Table 11 Serum TyG index of mice in each group

[0151]

[0152]

[0153] Note: ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0154] 2.4 Effects of different high-fat diets on aortic valve mass in mice

[0155] The effects of different high-fat diets on aortic valve plaques in mice are shown in Table 12. As shown in Table 12, no obvious plaques were found in the control group (0.31±0.30% to 0.44±0.27%); the positive rate of oil red staining in the traditional high-fat diet group increased significantly to 21.77±5.60% at 12 weeks (P<0.05 compared with the control group), and remained at 21.73±1.98%-21.97±1.38% at 16-20 weeks; the positive rate of oil red staining in the modified high-fat diet group increased significantly to 21.77±5.60% at 12 weeks (P<0.05 compared with the control group), and remained at 21.73±1.98%-21.97±1.38% at 16-20 weeks .... The positive rate of oil red staining in the feed group reached 36.37±15.13% at 12 weeks (P<0.05 compared with the control group, and increased by 67.0% compared with the traditional group), and maintained at 35.16±11.39%-35.79±9.65% at 16-20 weeks (P<0.01 compared with the control group), which was always the highest among the three groups and significantly increased compared with the traditional group (P<0.01 compared with the control group and the traditional high-fat feed at 20 weeks, increased by 62.9%).

[0156] The experimental results show that a high-fat diet can induce a significant increase in this indicator, and the effect of the modified high-fat feed is stronger, with an earlier starting time and a larger increase, and its impact on this indicator is significantly greater than that of the traditional high-fat feed.

[0157] Table 12 Average aortic plaque positive rate of mice in each group (plaque area / total blood vessel area)

[0158]

[0159] Note: # indicates P < 0.05 compared with the control group, ## indicates P < 0.01 compared with the control group, ** indicates P < 0.01 compared with the traditional feed.

[0160] In summary, the modified high-fat diet is more likely to induce increased blood pressure, lipid metabolism disorders and insulin resistance in mice than the traditional high-fat diet. Its destructive effect on metabolic homeostasis is earlier, more severe and more comprehensive, which may be related to enhanced inflammation or oxidative stress response.

[0161] The above is only an embodiment of the present invention, and common knowledge such as the specific technical solutions or characteristics in the solution is not described in detail here. It should be pointed out that for those skilled in the art, without departing from the technical solution of the present invention, several variations and improvements can be made, which should also be regarded as the scope of protection of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the description can be used to interpret the content of the claims.

Claims

1. A high-fat feed composition for metabolic syndrome in mice, characterized in that: By weight, it includes 185-205 parts of casein VTF, 1.5-4.5 parts of DL-methionine, 40-60 parts of corn starch, 85-115 parts of maltodextrin, 350-370 parts of sucrose, 40-60 parts of cellulose, 190-210 parts of anhydrous milk fat, 8-12 parts of corn oil, 0.04 parts of ethoxyquin, 30-40 parts of mineral mixture S10001AIN-76A, 35-45 parts of calcium carbonate, 8-12 parts of vitamin mixture 40060-D and 1.5 parts of cholesterol.

2. A high-fat feed composition for metabolic syndrome in mice according to claim 1, characterized in that: By weight, it includes 195 parts of casein VTF, 3 parts of DL-methionine, 50 parts of corn starch, 100 parts of maltodextrin, 341.46 parts of sucrose, 50 parts of cellulose, 200 parts of anhydrous milk fat, 10 parts of corn oil, 0.04 parts of ethoxyquin, 35 parts of mineral mixture S10001AIN-76A, 39 parts of calcium carbonate, 10 parts of vitamin mixture 40060-D and, 1.5 parts of cholesterol.

3. The preparation process of a high-fat feed composition for metabolic syndrome in mice according to claim 1, characterized in that: The following steps are involved: S1. Raw material pretreatment Solid raw material grinding: casein VTF, corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A and cholesterol are ground separately; Trace ingredient premix: DL-methionine, vitamin mixture 40060-D, and ethoxyquin are trace ingredients. Mix the trace ingredients with corn starch. Fat processing: anhydrous milk fat is first melted and then filtered to remove impurities; corn oil is checked for deterioration; S2. Ingredients According to the content of each raw material of the high-fat feed composition, the required raw materials are weighed by mass, and the weighed raw materials are placed separately for subsequent mixing operations; S3, Mixing Primary mixing: casein VTF, corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A and cholesterol are put into a mixer for primary mixing; Adding premix and fat: Add the premix of DL-methionine, vitamin mixture 40060-D, and ethoxyquin prepared in step S1, and the melted anhydrous milk fat and corn oil into a mixer and continue mixing; S4. Granulation Conditioning: Steam is introduced into the powdered feed obtained by mixing in step S3 to condition the feed, thereby gelatinizing the corn starch and denaturing the casein VTF, thereby improving the palatability and formability of the high-fat feed composition; Compression: The conditioned feed is fed into the pellet mill for compression; Cooling: Cool the hot pellet feed obtained by pressing to prevent the feed from getting moldy and deteriorating during storage; S5, sterilization, post-processing Sterilize the pellet feed prepared in step S4, and screen and package the sterilized pellet feed in sequence; S6. Finished product inspection and storage Quality inspection: Conduct sensory testing, nutrient analysis, and microbiological testing on finished feed to ensure that the feed quality meets experimental requirements; Storage: Store qualified feed in a cool, dry, and ventilated warehouse, avoid direct sunlight and storage with toxic and hazardous substances. Regularly check the quality of inventory feed, follow the first-in-first-out principle, and ensure that the feed is used within the shelf life.

4. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 3, characterized in that: In S1, a hammer mill is used to grind the solid raw materials into 80-100 meshes; and the hammer mill is cleaned after each solid raw material is ground to avoid cross contamination of the solid raw materials.

5. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 4, characterized in that: The mass ratio of the trace ingredients to the corn starch is 1:10-20, and the trace ingredients and the corn starch are mixed with a mixer for 5-8 minutes to ensure that the trace ingredients are evenly dispersed in the high-fat feed.

6. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 5, characterized in that: Melt the anhydrous milk fat by heating in a water bath at a temperature of 40-50°C.

7. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 3, characterized in that: In S2, each raw material is weighed using a high-precision electronic balance or an electronic batching scale.

8. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 3, characterized in that: In S3, casein VTF, corn starch, maltodextrin, sucrose, cellulose, calcium carbonate, mineral mixture S10001AIN-76A and cholesterol are mixed using a horizontal ribbon mixer, and the speed of the horizontal ribbon mixer is 40 to 60 rpm, and the mixing time is 5 to 8 minutes; when adding the premix and fat, the speed of the horizontal ribbon mixer is adjusted to 60 to 80 rpm, and the mixing time is 10 to 15 minutes to ensure that the fat evenly wraps the raw material particles and the trace components are fully dispersed, and the coefficient of variation of the mixing uniformity is ≤5%.

9. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 3, characterized in that: In S4, the steam pressure during conditioning is 0.2-0.4 MPa, the conditioning temperature is 75-85° C., the conditioning time is 2-3 minutes, and the moisture content of the feed after conditioning is 16%-18%.

10. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 9, characterized in that: The diameter of the die hole of the pellet mill is 1 to 2 mm, and the pressure of the pellet mill roller is 0.5 to 0.8 MPa to ensure that the feed passes through the die hole smoothly to form pellets.

11. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 10, characterized in that: The hot pellet feed is cooled by a counter-current cooler for 8 to 12 minutes. After cooling, the temperature of the pellets drops to room temperature ±5°C, and the moisture content of the feed pellets drops to 12% to 14%.

12. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 3, characterized in that: In S5, pellet feed is sterilized using cobalt-60 or electron beam irradiation equipment; pellet feed is packaged in aluminum foil bags or food-grade plastic sealed bags and divided according to experimental specifications; nitrogen is injected into the bag during packaging to remove air from the bag and delay feed oxidation, and the feed name, formula composition, production date, shelf life, storage conditions, applicable experiment type and sterilization method are clearly marked on the package.

13. The process for preparing a high-fat feed composition for metabolic syndrome in mice according to claim 3, characterized in that: In S6, sensory testing of finished feed includes color, odor, and particle size testing; nutritional analysis includes the determination of crude protein, crude fat, crude fiber, moisture, ash, cholesterol, and methionine content; microbial testing includes the detection of total colony count, E. coli count, and mold count; the storage temperature is 10-25°C, and the relative humidity is ≤65%.

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