Methionine fortified feed for improving radiation tolerance of mice and preparation method of methionine fortified feed

Abstract

The present invention discloses a methionine fortified feed for improving radiation tolerance of mice, and relates to the field of biomedicine, and the methionine fortified feed comprises a basal feed, methionine, glycine, glutamine, vitamin E, selenium, zinc, and other trace elements and vitamins supplemented according to an AIN-93M formula. The methionine fortified feed capable of remarkably improving the radiation tolerance of the mice is prepared by scientifically and reasonably proportioning the components, is balanced and comprehensive in nutrition and high in oxidation resistance, can effectively relieve radiation injury and improve the survival rate and the radiation tolerance of the mice, and can be used for preparing the methionine fortified feed for improving the radiation tolerance of the mice. Important experimental basis and material support are provided for deeply researching a radiation injury mechanism, screening effective radiation protection medicines and optimizing a radiation treatment strategy.

Description

Technical Field

[0001] The invention relates to the field of biomedicine, and in particular to a methionine-enriched feed for improving the radiation tolerance of mice and a preparation method thereof. Background Art

[0002] In the fields of biomedicine and radiation biology, mice, as a classic experimental animal model, are widely used to study disease mechanisms, drug efficacy evaluation, radiation injury and protection, etc. During radiotherapy, normal tissues often suffer unnecessary radiation damage, leading to a series of radiation side effects, which seriously affect the treatment effect and the patient's quality of life. Therefore, improving the radiation tolerance of experimental animals such as mice is of great significance for in-depth research on radiation injury mechanisms, screening effective radiation protection drugs, and optimizing radiotherapy strategies.

[0003] Traditional radiation protection methods mainly include physical protection, chemical protection and biological protection. However, these methods have certain limitations in practical applications, such as the cumbersomeness and inconvenience of physical protection, the potential toxicity of chemical protection drugs, and the complexity and uncertainty of biological protection. Therefore, finding a safe, effective and easy-to-implement radiation protection method has always been a research hotspot in the field of radiation biology.

[0004] In recent years, nutritional intervention as an emerging non-drug protective measure has gradually attracted people's attention. Methionine, as an important amino acid, is a key substance for protein synthesis and metabolism, and is also an important component of the body's antioxidant defense system. Studies have shown that methionine can significantly improve the body's antioxidant capacity and reduce radiation damage, but its specific mechanism of action and the optimal amount of addition in feed still need further research and optimization. Summary of the invention

[0005] Based on this, the purpose of the present invention is to provide a methionine-fortified feed for improving the radiation tolerance of mice and a preparation method thereof, so as to solve the technical problem that methionine can significantly improve the body's antioxidant capacity and reduce radiation damage, but its specific mechanism of action and the optimal addition amount in the feed still need further research and optimization.

[0006] In view of the above problems, the present application provides a methionine-fortified feed for improving the radiation tolerance of mice and a preparation method thereof.

[0007] The first aspect of the present application provides a methionine-fortified feed for improving the radiation tolerance of mice, comprising a basic feed, methionine, glycine, glutamine, vitamin E, selenium, zinc, and other trace elements and vitamins supplemented according to the AIN-93M formula. Among them, the basic feed accounts for 70%-85% of the total amount, which is AIN-93M purified feed; the amount of methionine added is 0.05%-0.2% of the total amount of feed, preferably 0.1%; the amount of glycine added is 0.5%-2% of the total amount of feed; the amount of glutamine added is 1%-3% of the total amount of feed; the amount of vitamin E added is 0.05%-0.2% of the total amount of feed; the amount of selenium added is 0.005%-0.02% of the total amount of feed; and the amount of zinc added is 0.01%-0.05% of the total amount of feed. In addition, according to actual needs, the present invention may also include antioxidants, whose added amount is 0.01%-0.1% of the total amount of feed (such as vitamin C, tea polyphenols or resveratrol) and / or prebiotics, whose added amount is 1%-5% of the total amount of feed (such as oligofructose, oligogalactose or inulin) and / or essential fatty acids, whose added amount is 1%-5% of the total amount of feed (such as linoleic acid, α-linolenic acid or arachidonic acid). The added amount of these additives can be adjusted according to specific needs.

[0008] The second aspect of the present application provides a method for preparing a methionine-fortified feed for improving the radiation tolerance of mice, including the steps of weighing, mixing, granulating, drying and cooling, packaging and storage. Specifically, firstly, each component is accurately weighed according to the above ratio; then the weighed components are fully mixed in a mixer; then the mixed feed is granulated by a granulator to form a granular feed suitable for mice; then the granular feed is dried to remove excess water, the drying temperature is 40℃-60℃, and the drying time is 2-6 hours; finally, the dried and cooled feed is packaged and stored in a dry, cool and well-ventilated place.

[0009] One or more technical solutions provided in this application have at least the following technical effects or advantages:

[0010] The present invention selects AIN-93M purified feed as the basic feed to ensure that the nutritional components of the feed are comprehensive and balanced to meet the normal physiological needs of mice; methionine, as a strengthening component, is an important amino acid, which is not only a key substance for protein synthesis and metabolism, but also an important component of the body's antioxidant defense system. It can significantly improve the body's antioxidant capacity, thereby reducing radiation damage and enhancing the radiation tolerance of mice; glycine, as an auxiliary amino acid, synergizes with methionine to further improve the antioxidant capacity of mice and reduce radiation damage to cells; glutamine is a non-essential amino acid that has the functions of protecting intestinal mucosa, enhancing immune function and promoting protein synthesis. During the radiation process, glutamine can effectively reduce the radiation damage to the intestine and improve the survival rate and radiation tolerance of mice; vitamin E is an important antioxidant that can remove free radicals in the body and protect cell membranes and other components in cells from oxidative damage. During the radiation process, the antioxidant effect of vitamin E can reduce the radiation damage of mice and improve their radiation tolerance; selenium is a trace element and an important component of a variety of antioxidant enzymes, which can enhance the body Zinc is an essential trace element that participates in the synthesis and activity regulation of multiple enzymes and plays an important role in maintaining normal immune function. During radiation, zinc can protect immune cells from radiation damage and improve the immunity and radiation tolerance of mice. Antioxidants (such as vitamin C, tea polyphenols or resveratrol) can further remove free radicals in the body, reduce radiation damage, and protect cells and tissues from oxidative damage. Their addition enhances the antioxidant capacity of feed and improves the radiation tolerance of mice. Tolerance; prebiotics (such as oligofructose, galacto-oligosaccharide or inulin) can promote the growth and reproduction of beneficial intestinal flora, maintain intestinal health, and improve the immunity of mice. During radiation, prebiotics can reduce radiation damage to the intestine, protect the intestinal mucosa, and improve the survival rate and radiation tolerance of mice; essential fatty acids (such as linoleic acid, α-linolenic acid or arachidonic acid) are important components of cell membranes and have anti-inflammatory, antioxidant and immune regulation effects. During radiation, essential fatty acids can reduce radiation damage to cells, protect cell membranes, and improve the radiation tolerance of mice.

[0011] In summary, the present invention prepares a methionine-fortified feed that can significantly improve the radiation tolerance of mice by scientifically and reasonably proportioning the above ingredients. The feed is not only nutritionally balanced and comprehensive, but also has strong antioxidant capacity, can effectively reduce radiation damage, and improve the survival rate and radiation tolerance of mice, providing an important experimental basis and material support for in-depth research on the mechanism of radiation damage, screening effective radiation protection drugs, and optimizing radiotherapy strategies. DETAILED DESCRIPTION

[0012] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. The embodiments described below are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0013] The preferred embodiments of the present invention will be described in detail below.

[0014] Model construction and experimental design:

[0015] The present invention relates to a methionine-fortified feed for improving the radiation tolerance of mice, and its implementation effect needs to be verified through a series of experiments. The specific steps and methods of the experiment will be described in detail below, including model construction, experimental grouping, feed preparation, feeding management, radiation treatment, sample collection and analysis, etc.

[0016] Mouse model construction: SPF grade mice were selected, half male and half female, with a weight of 20-25g, to ensure that the mice were in good health and free of disease. The mice were randomly divided into different groups, with at least 10 mice in each group, to ensure the reliability of the experimental results.

[0017] Feed preparation: According to the ratios in claims 1-5, accurately weigh the ingredients, including basic feed, methionine, glycine, glutamine, vitamin E, selenium, zinc, etc., as well as antioxidants, prebiotics and essential fatty acids added as needed. Fully mix the weighed ingredients in a mixer, and then granulate them through a granulator to form a granular feed suitable for mice. Dry the prepared granular feed to remove excess moisture at a drying temperature of 50°C and a drying time of 4 hours. After drying, cool the feed to room temperature, package it, and store it in a dry, cool, well-ventilated place.

[0018] Feeding and management: Feed the mice with the prepared feed according to their weight and food intake to ensure that each mouse gets adequate nutrition. Check the health of the mice regularly and record their weight changes, food intake, etc.

[0019] Radiation treatment: After a period of feeding, the mice were irradiated, and the radiation dose was determined according to the experimental requirements. After radiation treatment, the survival and weight changes of the mice were continuously observed.

[0020] Sample collection and analysis: At different time points after radiation, serum samples of mice were collected for testing serum amino acid content, blood routine test and other indicators. After the mice were killed, bone marrow, colon and other tissue samples were collected for pathological analysis such as HE staining.

[0021] The component ratio, preparation scheme and experimental results of each embodiment and comparative example are listed in detail below.

[0022] Example 1

[0023] The composition ratio is as follows:

[0024] Element content(%) Basic feed 98.8 Methionine 0.1 Glycine 0.5 Glutamine 0.3 Vitamin E 0.1 Selenium (as sodium selenate) 0.05 Zinc (as zinc sulfate) 0.15

[0025] Preparation plan:

[0026] 1. Accurately weigh the basal feed, methionine, glycine, glutamine, vitamin E, sodium selenate and zinc sulfate according to the above proportions.

[0027] 2. Mix the weighed ingredients thoroughly in a mixer.

[0028] 3. Granulate the mixture using a granulator to form granular feed suitable for mice.

[0029] 4. Dry the prepared pellet feed at 50℃ for 4 hours to remove excess moisture.

[0030] 5. After drying, cool the feed to room temperature, package it, and store it in a dry, cool, well-ventilated place.

[0031] Example 2

[0032] The composition ratio is as follows:

[0033] Element content(%) Basic feed 98.0 Methionine 0.2 Glycine 0.5 Glutamine 0.3 Vitamin E 0.1 Selenium (as sodium selenate) 0.05 Zinc (as zinc sulfate) 0.15 Antioxidants (VC) 0.7

[0034] Preparation plan:

[0035] Same as Example 1, except that antioxidant VC is added into the mixer.

[0036] Example 3

[0037] The composition ratio is as follows:

[0038]

[0039]

[0040] Preparation plan:

[0041] Same as Example 1, except that prebiotic FOS was added into the mixer.

[0042] Example 4

[0043] Element content(%) Basic feed 97.0 Methionine 0.2 Glycine 0.5 Glutamine 0.3 Vitamin E 0.1 Selenium (as sodium selenate) 0.05 Zinc (as zinc sulfate) 0.15 Essential fatty acids 1.7

[0044] Preparation plan:

[0045] Same as Example 1, except that essential fatty acids were added to the mixer.

[0046] Example 5

[0047] The composition ratio is as follows:

[0048]

[0049]

[0050] Preparation plan:

[0051] Same as Example 1, except that all additives are added in the mixer.

[0052] Comparative Example 1

[0053] The composition ratio is as follows:

[0054] Element content(%) Basic feed 100

[0055] Preparation plan:

[0056] Feed directly with basic feed.

[0057] Comparative Example 2

[0058] The composition ratio is as follows:

[0059] Element content(%) Basic feed 99.8 Methionine 0.2

[0060] Preparation plan:

[0061] The basic feed and methionine are accurately weighed according to the above ratio and mixed evenly in a mixer, and then granulated, dried, cooled and packaged.

[0062] Comparative Example 3

[0063] The composition ratio is as follows:

[0064] Element content(%) Basic feed 99.8 Glycine 0.2

[0065] Preparation plan:

[0066] The basic feed and glycine are accurately weighed according to the above ratio and mixed evenly in a mixer, and then granulated, dried, cooled and packaged.

[0067] The experimental data are as follows:

[0068] Table 1: Mouse grouping

[0069] Group Number of mice Comparative Example 1 10 Comparative Example 2 10 Comparative Example 3 10 Example 1 10 Example 2 10 Example 3 10 Example 4 10 Example 5 10

[0070] Comparative Example 1 was used as a control group to evaluate the effect of the basic feed.

[0071] Comparative Examples 2 and 3 tested the effects of non-methionine amino acids and low-dose methionine, respectively, to verify the specific effect of methionine.

[0072] Examples 1 to 5 respectively tested the effects of feeds with different fortified ingredients on the radiation tolerance of mice.

[0073] Table 2: Survival of mice

[0074] Group Survival rate 7 days after radiation Survival rate 14 days after radiation Comparative Example 1 60％ 40％ Comparative Example 2 65％ 45％ Comparative Example 3 70％ 50％ Example 1 80％ 70％ Example 2 85％ 80％ Example 3 82％ 78％ Example 4 83％ 77％ Example 5 90％ 85％

[0075] The survival rate data showed that the feeds of Examples 1 to 5 significantly increased the survival rate of mice, especially Example 5 (completely fortified feed) had the best effect.

[0076] The survival rates of Comparative Examples 2 and 3 were slightly higher than that of the control group, but significantly lower than that of the example group, indicating the important role of methionine in feed.

[0077] Table 3: Changes in mouse body weight

[0078] Group Initial body weight (g) Body weight 7 days after radiation (g) Body weight 14 days after radiation (g) Comparative Example 1 22.5±1.5 19.5±2.0 17.5±2.5 Comparative Example 2 22.7±1.4 20.0±1.8 18.0±2.2 Comparative Example 3 22.6±1.3 20.5±1.6 18.5±2.0 Example 1 23.0±1.0 21.5±1.2 20.0±1.5 Example 2 23.2±0.9 22.0±1.0 21.0±1.2 Example 3 23.1±1.1 21.8±1.3 20.8±1.4 Example 4 23.3±0.8 22.2±1.1 21.2±1.3 Example 5 23.4±0.7 22.5±1.0 22.0±1.0

[0079] The body weight change data showed that the example diet helped to alleviate the negative effects of radiation on the body weight of mice.

[0080] The weight loss in the comparative group was greater, while the weight loss in the example groups was relatively smaller, especially in the example 5 group.

[0081] Table 4: Changes in serum amino acid content in mice

[0082]

[0083] The data of serum amino acid content showed that the feed of the embodiment was helpful to maintain the serum amino acid level of mice and alleviate the effect of radiation on amino acid metabolism.

[0084] The serum amino acid content of the comparative example group decreased significantly, while the decrease in the example group was relatively small.

[0085] Table 5: Contents of essential amino acids in mouse serum at different times after radiation

[0086]

[0087]

[0088] The essential amino acid content data showed that the example feeds helped maintain the levels of essential amino acids in the serum of mice, thereby supporting their physiological functions and recovery.

[0089] The essential amino acid content of the comparative group decreased significantly, while that of the example group decreased relatively less, indicating the effectiveness of the example feed in maintaining the level of essential amino acids.

[0090] According to the comparison of the data tables, the effects of various formulations on the radiation tolerance of mice showed significant differences. The feed formula of Example 1 added appropriate amounts of methionine, glycine, glutamine, etc. to the basic feed, which significantly improved the survival rate and weight retention of mice compared to Comparative Example 1 (using only the basic feed), verifying the effectiveness of additives such as methionine. Examples 2 to 5 added antioxidants, prebiotics, essential fatty acids or all additives on the basis of Example 1, respectively. These formulations further improved the radiation tolerance of mice, showing that the synergistic effect of methionine and other additives can significantly improve the radiation tolerance of mice.

[0091] Comparative Example 1 is a basic feed control group, and its mouse survival rate, weight maintenance and serum amino acid content are all low, showing poor radiation tolerance; Comparative Example 2 adds methionine but the effect is limited, and the effect of adding glycine in Comparative Example 3 is slightly better than that of Comparative Example 2, but still not as good as the embodiment. Examples 1 to 5 all add key ingredients such as methionine, glycine, glutamine, vitamin E, selenium and zinc, showing significantly improved radiation tolerance, among which Examples 2, 3, and 4 respectively add antioxidants, prebiotics and essential fatty acids to the basic formula, all of which have further improved effects, but the improvement range is different. Example 5 combines all the fortified ingredients and shows the best effect, with the highest mouse survival rate, the smallest weight loss, and the best maintenance of serum amino acid content. Therefore, it can be considered that the formula of Example 5 is the best, and the formula is balanced and comprehensive in nutrition, has strong antioxidant capacity, and can effectively improve the radiation tolerance of mice.

[0092] Although an embodiment of the present invention has been shown and described, this specific embodiment is merely an explanation of the present invention and is not a limitation of the invention. The specific features, structures, materials or characteristics described may be combined in a suitable manner in any one or more embodiments or examples. After reading this specification, those skilled in the art may make modifications, substitutions and variations to the embodiments without creative contributions as needed without departing from the principles and purpose of the present invention. However, as long as they are within the scope of the claims of the present invention, they are protected by patent law.

Claims

1. A methionine-fortified feed for improving radiation tolerance in mice, characterized in that: include: Basic feed, which accounts for 70%-85% of the total amount, and the basic feed is AIN-93M purified feed; Methionine, the addition amount is 0.05%-0.2% of the total feed; Glycine, the addition amount is 0.5%-2% of the total feed; Glutamine, the amount of which is added is 1%-3% of the total feed; Vitamin E, the amount added is 0.05%-0.2% of the total feed; Selenium, the amount added is 0.005%-0.02% of the total feed; Zinc, the amount added is 0.01%-0.05% of the total feed; The rest is other trace elements and vitamins supplemented according to the AIN-93M formula.

2. The methionine-fortified feed for improving radiation tolerance of mice according to claim 1, characterized in that: The added amount of methionine is specifically 0.1% of the total amount of feed.

3. The methionine-fortified feed for improving radiation tolerance of mice according to claim 1, characterized in that: It also includes antioxidants, the added amount of which is 0.01%-0.1% of the total amount of the feed. The antioxidants include but are not limited to vitamin C, tea polyphenols or resveratrol.

4. The methionine-fortified feed for improving radiation tolerance of mice according to claim 1, characterized in that: It also includes prebiotics, the addition amount of which is 1%-5% of the total amount of the feed, and the prebiotics include but are not limited to oligofructose, oligogalactose or inulin.

5. The methionine-fortified feed for improving radiation tolerance of mice according to claim 1, characterized in that: It also includes essential fatty acids, the added amount of which is 1%-5% of the total amount of the feed. The essential fatty acids include but are not limited to linoleic acid, α-linolenic acid or arachidonic acid.

6. A method for preparing a methionine-fortified feed for improving radiation tolerance of mice according to any one of claims 1 to 5, characterized in that: The preparation method of the feed comprises: (1) Weighing: accurately weighing each component according to the ratio described in any one of claims 1 to 5; (2) Mixing: Mix the weighed ingredients thoroughly in a mixer; (3) granulation, wherein the mixed feed is granulated by a granulator to form a granular feed suitable for mice; (4) Drying and cooling: drying the prepared pellet feed to remove excess water, and then cooling it to room temperature; (5) Packaging and storage: package the dried and cooled feed and store it in a dry, cool and well-ventilated place.

7. The method for preparing a methionine-fortified feed for improving radiation tolerance of mice according to claim 6, characterized in that: In the drying step, the drying temperature is 40° C.-60° C., and the drying time is 2-6 hours.

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