CN109007397A - A kind of fodder compound and preparation method thereof that can improve mouse heart morphosis and function - Google Patents
A kind of fodder compound and preparation method thereof that can improve mouse heart morphosis and function Download PDFInfo
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- CN109007397A CN109007397A CN201810964657.8A CN201810964657A CN109007397A CN 109007397 A CN109007397 A CN 109007397A CN 201810964657 A CN201810964657 A CN 201810964657A CN 109007397 A CN109007397 A CN 109007397A
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Abstract
Description
技术领域technical field
本发明涉及实验动物营养学领域，尤其是一种适合于Omega3过表达小鼠生长并维持典型模型特征且对其心脏形态结构与功能无副作用的饲料组合物及其制备方法。The invention relates to the field of experimental animal nutrition, in particular to a feed composition suitable for the growth of Omega3 overexpressed mice and maintaining typical model characteristics without side effects on the morphology, structure and function of the heart and a preparation method thereof.
背景技术Background technique
目前，心血管疾病已成为全球人类的“头号杀手”。ω-3多不饱和脂肪酸(ω-3PUFAs)对人类及其他哺乳动物的正常发育和维持健康极其重要，且在人类心血管疾病的预防与治疗中发挥着突出的作用。但是ω-3多不饱和脂肪酸在大多数动物体内不能合成，只能从食物中补充。目前科学家已在人类饮食中添加ω-3PUFAs做了许多研究，获得显著的效果，很多富含ω-3PUFAs产品已成为保健品甚至药物被临床广泛应用，但是外源性添加ω-3PUFAs仍存在许多问题，如膳食结构、环境等混杂因素的干扰。因此，亟需寻找一种可以排除膳食结构等外部因素干扰的方法来研究ω-3PUFAs对心血管疾病的防治作用及其机制。At present, cardiovascular disease has become the "number one killer" of human beings in the world. ω-3 polyunsaturated fatty acids (ω-3PUFAs) are extremely important to the normal development and maintenance of health of humans and other mammals, and play a prominent role in the prevention and treatment of human cardiovascular diseases. However, omega-3 polyunsaturated fatty acids cannot be synthesized in most animals and can only be supplemented from food. At present, scientists have done a lot of research on adding ω-3PUFAs to the human diet, and achieved remarkable results. Many products rich in ω-3PUFAs have become health products and even drugs have been widely used clinically, but there are still many ω-3PUFAs added exogenously. Problems, such as the interference of confounding factors such as dietary structure and environment. Therefore, it is urgent to find a method that can exclude the interference of external factors such as dietary structure to study the prevention and treatment effect and mechanism of ω-3PUFAs on cardiovascular diseases.
Omega3过表达小鼠可将体内多余的ω-6PUFAs转化为ω-3PUFAs，从而使动物的组织和器官中富含ω-3PUFAs，并且组织中的ω-6/ω-3PUFAs比例趋于均衡。目前Omega3过表达小鼠因不受膳食结构等外部因素的干扰，已成全球学者研究ω-3PUFAs或ω-3/ω-6PUFAs比例对心血管疾病保护作用的理想动物模型。但Omega3过表达小鼠没有专用的饲料喂养，目前研究者均给予温和饲料(含有10％左右的脂肪)进行科学研究，但给予温和饲料后引起心脏形态结构与功能改变的现象引起研究者高度关注，如心脏腔室明显扩大，该缺陷给研究内源性ω-3PUFAs对心脏的保护作用机制带来了一定的局限性。因此，亟待研制出一种既满足Omega3过表达小鼠营养需求与维持模型特征的同时，又对该小鼠心脏形态结构与功能无副作用的饲料组合物。Omega3 overexpression mice can convert excess ω-6 PUFAs into ω-3 PUFAs in the body, so that the tissues and organs of the animal are rich in ω-3 PUFAs, and the ratio of ω-6/ω-3 PUFAs in the tissues tends to be balanced. At present, Omega3 overexpression mice are not disturbed by external factors such as dietary structure, and have become an ideal animal model for scholars around the world to study the protective effect of ω-3 PUFAs or ω-3/ω-6 PUFAs ratio on cardiovascular disease. However, Omega3 overexpression mice do not have a special feed for feeding. At present, researchers are giving mild feed (containing about 10% fat) for scientific research, but the phenomenon of changes in heart morphology and function after giving mild feed has attracted great attention from researchers. , such as the obvious expansion of the heart chamber, this defect has brought certain limitations to the study of the protective mechanism of endogenous ω-3PUFAs on the heart. Therefore, it is urgent to develop a feed composition that not only satisfies the nutritional requirements of Omega3 overexpressed mice and maintains the characteristics of the model, but also has no side effects on the morphology and function of the mouse heart.
发明内容Contents of the invention
本发明的目的在于克服上述现有技术的不足之处而提供一种适合Omega3过表达小鼠生长并维持模型特征且对心脏形态结构与功能无副作用的饲料组合物。The purpose of the present invention is to overcome the shortcomings of the above-mentioned prior art and provide a feed composition suitable for the growth of Omega3 overexpression mice and maintaining the characteristics of the model without side effects on the morphology, structure and function of the heart.
本发明的另一目的还在于提供适合Omega3过表达小鼠生长并维持模型特征且对心脏形态结构与功能无副作用的饲料组合物的制备方法，该制备方法操作简单，适于工业化生产。Another object of the present invention is to provide a preparation method of a feed composition that is suitable for the growth of Omega3 overexpressed mice and maintains the characteristics of the model, and has no side effects on the shape, structure and function of the heart. The preparation method is simple to operate and suitable for industrial production.
为实现上述目的，本发明采取的技术方案为：一种可以改善Omega3过表达小鼠心脏形态结构的饲料组合物，所述饲料组合物的组分包含玉米粉、麸皮、豆粕、酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、植物油、动物油脂、混合矿物质、混合维生素和纤维素。In order to achieve the above object, the technical solution adopted by the present invention is: a feed composition that can improve the heart morphology of Omega3 overexpression mice, the components of the feed composition include corn flour, bran, soybean meal, casein, Sugars, Lysine, Methionine, Cystine, Vegetable Oils, Animal Fats, Mixed Minerals, Mixed Vitamins and Cellulose.
作为本发明所述可以改善Omega3过表达小鼠心脏形态结构与功能的饲料组合物的实施方式，所述饲料组合物中各组分的重量份数分别为：玉米粉31～32份、麸皮7～8份、豆粕8.5～10份、酪蛋白4～6份、糖类8～11份、赖氨酸11～13份、蛋氨酸5～6份、胱氨酸2～3份、植物油3～4.5份、动物油脂2～4份、混合矿物质3～5份、混合维生素0.8～1.5份、纤维素4～5份。As an embodiment of the feed composition that can improve the morphological structure and function of the heart of Omega3 overexpression mice described in the present invention, the parts by weight of each component in the feed composition are: 31-32 parts of corn flour, 32 parts of bran 7-8 parts, 8.5-10 parts of soybean meal, 4-6 parts of casein, 8-11 parts of sugar, 11-13 parts of lysine, 5-6 parts of methionine, 2-3 parts of cystine, 3-3 parts of vegetable oil 4.5 parts, 2-4 parts of animal fat, 3-5 parts of mixed minerals, 0.8-1.5 parts of mixed vitamins, 4-5 parts of cellulose.
作为本发明所述可以改善Omega3过表达小鼠心脏形态结构与功能的饲料组合物的优选实施方式，所述饲料组合物中各组分的重量份数分别为：玉米粉31.4份、麸皮7.6份、豆粕9份、酪蛋白5份、糖类10份、赖氨酸12份、蛋氨酸5.6份、胱氨酸2.4份、植物油3.4份、动物油脂2.6份、混合矿物质5份、混合维生素1.2份、纤维素4份。As a preferred embodiment of the feed composition that can improve the morphological structure and function of the heart of Omega3 overexpression mice described in the present invention, the parts by weight of each component in the feed composition are respectively: 31.4 parts of corn flour, 7.6 parts of bran 9 parts of soybean meal, 5 parts of casein, 10 parts of sugar, 12 parts of lysine, 5.6 parts of methionine, 2.4 parts of cystine, 3.4 parts of vegetable oil, 2.6 parts of animal fat, 5 parts of mixed minerals, 1.2 parts of mixed vitamins 4 parts, 4 parts of cellulose.
作为本发明所述Omega3过表达小鼠饲料组合物的优选实施方式，所述植物油为玉米油、橄榄油、花生油、菜籽油、亚麻油中的至少一种。As a preferred embodiment of the Omega3 overexpression mouse feed composition of the present invention, the vegetable oil is at least one of corn oil, olive oil, peanut oil, rapeseed oil, and linseed oil.
作为本发明所述Omega3过表达小鼠饲料组合物的优选实施方式，所述动物油脂为猪油、牛油、鱼油中的至少一种。As a preferred embodiment of the Omega3 overexpression mouse feed composition of the present invention, the animal fat is at least one of lard, tallow, and fish oil.
作为本发明所述Omega3过表达小鼠饲料组合物的优选实施方式，所述混合矿物质包括碳酸氢钙、氯化钠、硫酸铁、硫酸锌、碘化钾及硫酸铜。As a preferred embodiment of the Omega3 overexpression mouse feed composition of the present invention, the mixed minerals include calcium bicarbonate, sodium chloride, iron sulfate, zinc sulfate, potassium iodide and copper sulfate.
作为本发明所述Omega3过表达小鼠专饲料组合物的优选实施方式，所述混合维生素包括维生素A、维生素D、维生素E、维生素K、维生素B1、维生素B2、维生素B6、维生素B12、盐酸、泛酸钙、叶酸及生物素。As a preferred embodiment of the Omega3 overexpression mouse feed composition of the present invention, the mixed vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B 1 , vitamin B 2 , vitamin B 6 , vitamin B 12. Hydrochloric acid, calcium pantothenate, folic acid and biotin.
作为本发明所述Omega3过表达小鼠饲料组合物的优选实施方式，所述纤维素为羧甲基纤维素、聚乙烯纤维素、木质素中的至少一种。As a preferred embodiment of the Omega3 overexpression mouse feed composition of the present invention, the cellulose is at least one of carboxymethyl cellulose, polyethylene cellulose, and lignin.
作为本发明所述Omega3过表达小鼠饲料组合物的优选实施方式，所述玉米粉、麸皮和豆粕的粒径均为80～100目。As a preferred embodiment of the Omega3 overexpression mouse feed composition of the present invention, the particle diameters of the corn flour, bran and soybean meal are all 80-100 mesh.
作为本发明所述Omega3过表达小饲料组合物的优选实施方式，所述糖类为乳糖。As a preferred embodiment of the Omega3 overexpression small feed composition of the present invention, the carbohydrate is lactose.
另外，本发明还提供了一种如上所述Omega3过表达小鼠饲料组合物的制备方法，所述方法包括如下步骤：In addition, the present invention also provides a kind of preparation method of above-mentioned Omega3 overexpression mouse feed composition, described method comprises the following steps:
(1)预混：将酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、混合矿物质、混合维生素及纤维素进行预混，获得预混料；(1) Premixing: premixing casein, sugars, lysine, methionine, cystine, mixed minerals, mixed vitamins and cellulose to obtain a premix;
(2)混料：将上述预混料与玉米粉、麸皮、豆粕混合均匀，再加入植物油、动物油以及水，混合均匀，获得混合料；(2) Mixing: Mix the above-mentioned premix with corn flour, bran, and soybean meal, then add vegetable oil, animal oil and water, and mix evenly to obtain the mixture;
(3)制粒：采用制粒机将所述混合料制成颗粒；(3) granulation: adopt the granulator to make the mixture into granules;
(4)烘干：将所述颗粒置于烘箱内烘干；(4) drying: the particles are placed in an oven to dry;
(5)灭菌：将烘干颗粒分装后进行灭菌得成品饲料。(5) Sterilization: the dried granules are subpackaged and then sterilized to obtain finished feed.
作为本发明所述Omega3过表达小鼠饲料组合物的制备方法的优选实施方式，所述步骤(1)预混前先将玉米、麸皮和豆粕分别投到粉碎机进行粉碎，过80～100目筛后，备用。As a preferred embodiment of the preparation method of the Omega3 overexpression mouse feed composition of the present invention, before the step (1) premixing, the corn, bran and soybean meal are respectively thrown into the pulverizer for pulverization, and after 80-100 After mesh sieving, set aside.
作为本发明所述Omega3过表达小鼠饲料组合物的制备方法的优选实施方式，所述步骤(3)中，颗粒的粒径为10mm～12mm，长度为25mm。As a preferred embodiment of the preparation method of the Omega3 overexpression mouse feed composition of the present invention, in the step (3), the particle size of the particles is 10mm-12mm, and the length is 25mm.
作为本发明所述Omega3过表达小鼠饲料组合物的制备方法的优选实施方式，所述步骤(4)中，烘箱的温度为70℃～85℃。As a preferred embodiment of the preparation method of the Omega3 overexpression mouse feed composition of the present invention, in the step (4), the temperature of the oven is 70°C-85°C.
作为本发明所述Omega3过表达小鼠饲料组合物的制备方法的优选实施方式，所述步骤(5)中，采用20kGy 60Co-γ射线照射进行灭菌。As a preferred embodiment of the preparation method of the Omega3 overexpression mouse feed composition of the present invention, in the step (5), 20kGy 60Co-γ rays are used for sterilization.
作为本发明所述Omega3过表达小鼠饲料组合物的制备方法的优选实施方式，所述方法还包括步骤(6)成品饲料的检测：将步骤(5)制得的成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合国家标准要求。将所述成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合小鼠营养配方标准(GB 14924.3－2001)和实验动物营养配方通用质量要求(GB 14924.1－2001)。As a preferred embodiment of the preparation method of the Omega3 overexpression mouse feed composition of the present invention, the method also includes the detection of step (6) finished feed: the finished product made in step (5) is subjected to nutritional components and hygienic indicators Inspection to ensure that the indicators of the finished feed meet the requirements of national standards. The finished product is tested for nutritional components and hygienic indicators to ensure that each indicator of the finished feed meets the nutritional formula standard for mice (GB 14924.3-2001) and the general quality requirements for nutritional formula for experimental animals (GB 14924.1-2001).
作为本发明所述Omega3过表达小鼠饲料组合物的制备方法的优选实施方式，在制备前，先对所有原料进行营养和卫生指标的检测，使得所示Omega3过表达小鼠饲料组合物中的各组分的微生物、重金属含量、农药残留等指标均符合实验动物营养配方卫生标准(GB14924.2－2001)。As a preferred embodiment of the preparation method of the Omega3 overexpression mouse feed composition of the present invention, before preparation, all raw materials are carried out to the detection of nutritional and hygienic indicators, so that the Omega3 in the shown Omega3 overexpression mouse feed composition The microorganisms, heavy metal content, pesticide residues and other indicators of each component are in line with the hygienic standard of nutritional formula for experimental animals (GB14924.2-2001).
本发明所述Omega3过表达小鼠饲料组合物，采用所述特定配方的组分制备而成，本发明所述Omega3过表达小鼠饲料组合物的配方组分、形态和气味适合啮齿类动物的习性和嗜好。本发明的Omega3过表达小鼠饲料组合物通过科学配方，在满足Omega3过表达小鼠生长的营养需求，保证其典型的模型特征的同时消除了给予温和型饲料(饲料中含10％左右的脂肪)后导致的心脏形态结构与功能改变的副作用，为精准研究内源性ω-3PUFAs对心血管疾病的保护作用机制奠定了基础。而且，本发明所述的Omega3过表达小鼠饲料组合物，营养均衡、维持模型特征且对心脏无副作用、无无机物(包括重金属)、农药残留污染以及极低含量的植物雌激素，防止喂养后动物出现机能改变(如氧化应激指标、血脂指标等)，从而排除植物雌激素干扰内源性ω-3PUFAs对心血管保护作用机制的研究。The Omega3 overexpression mouse feed composition of the present invention is prepared from the components of the specific formula, and the formula components, shape and smell of the Omega3 overexpression mouse feed composition of the present invention are suitable for rodents. habits and hobbies. The Omega3 overexpression mouse feed composition of the present invention meets the nutritional requirements of Omega3 overexpression mouse growth through scientific formulation, and eliminates the need to give mild feed (containing about 10% fat in the feed) while ensuring its typical model characteristics. ) The side effects of cardiac morphological structure and functional changes have laid a foundation for the precise study of the protective mechanism of endogenous ω-3 PUFAs on cardiovascular diseases. Moreover, the Omega3 overexpression mouse feed composition of the present invention has balanced nutrition, maintains model characteristics and has no side effects on the heart, no inorganic substances (including heavy metals), pesticide residue pollution and extremely low content of phytoestrogens, preventing feeding Afterwards, the animals showed functional changes (such as oxidative stress indicators, blood lipid indicators, etc.), thus excluding the research on the mechanism of phytoestrogens interfering with endogenous ω-3PUFAs on cardiovascular protection.
附图说明Description of drawings
图1为各组小鼠典型的心脏超声心动图(M型)的比较图。Fig. 1 is a comparison diagram of typical heart echocardiograms (M-mode) of mice in each group.
图2为各组小鼠左心室收缩/舒张末期内径(LVID；s和LVID；d)的比较图。Fig. 2 is a comparison diagram of left ventricular end-systolic/diastolic diameter (LVID; s and LVID; d) of mice in each group.
图3为各组小鼠心脏左心室射血分数(EF)和短轴缩短率(FS)的比较图。Fig. 3 is a comparison chart of left ventricular ejection fraction (EF) and fractional shortening (FS) of mice in each group.
图4为各组小鼠血清CK和LDH含量的比较图。Fig. 4 is a comparison chart of serum CK and LDH contents of mice in each group.
图5为各组小鼠心脏组织cTnT和cTnI含量的比较图。Fig. 5 is a comparison chart of the contents of cTnT and cTnI in heart tissue of mice in each group.
具体实施方式Detailed ways
为更好的说明本发明的目的、技术方案和有益效果，下面将结合附图和具体实施例对本发明作进一步说明。In order to better illustrate the purpose, technical solutions and beneficial effects of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.
实施例1Example 1
本发明Omega3过表达小鼠饲料组合物的一种实施例，本实施例所述Omega3过表达小鼠饲料组合物包含以下重量份的组分：玉米粉31份、麸皮8份、豆粕9.2份、酪蛋白6份、糖类11份、赖氨酸11份、蛋氨酸5.2份、胱氨酸3份、植物油3份、动物油脂4、混合矿物质3份、混合维生素1.5份、纤维素5份。An embodiment of the Omega3 overexpression mouse feed composition of the present invention, the Omega3 overexpression mouse feed composition described in this embodiment comprises the following components by weight: 31 parts of corn flour, 8 parts of bran, and 9.2 parts of soybean meal , 6 parts of casein, 11 parts of sugar, 11 parts of lysine, 5.2 parts of methionine, 3 parts of cystine, 3 parts of vegetable oil, 4 parts of animal fat, 3 parts of mixed minerals, 1.5 parts of mixed vitamins, 5 parts of cellulose .
本实施例中，所述糖类为乳糖；所述植物油为玉米油；所述动物油脂为猪油；所述混合矿物质包括碳酸氢钙、氯化钠、硫酸铁、硫酸锌、碘化钾及硫酸铜；所述混合维生素包括维生素A、维生素D、维生素E、维生素K、维生素B1、维生素B2、维生素B6、维生素B12、盐酸、泛酸钙、叶酸及生物素；所述纤维素为羧甲基纤维素。In this embodiment, the saccharide is lactose; the vegetable oil is corn oil; the animal fat is lard; the mixed minerals include calcium bicarbonate, sodium chloride, iron sulfate, zinc sulfate, potassium iodide and sulfuric acid Copper; the mixed vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B 1 , vitamin B 2 , vitamin B 6 , vitamin B 12 , hydrochloric acid, calcium pantothenate, folic acid and biotin; the cellulose is Carboxymethylcellulose.
本实施例所述Omega3过表达小鼠饲料组合物采用如下方法制备而成：The Omega3 overexpression mouse feed composition described in this example is prepared by the following method:
(1)粉碎：将玉米、麸皮、豆粕分别投到粉碎机粉碎，过80～100目，备用；(1) Pulverization: Put corn, bran and soybean meal into a pulverizer for pulverization, pass through 80-100 meshes, and set aside;
(2)预混：将酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、混合矿物质、混合维生素及纤维素进行预混，获得预混料；(2) Premixing: premixing casein, sugars, lysine, methionine, cystine, mixed minerals, mixed vitamins and cellulose to obtain a premix;
(3)混料：将上述预混料与玉米粉、麸皮、豆粕混合均匀，再加入植物油、动物油以及水，混合均匀，获得混合料；(3) Mixing: Mix the above-mentioned premix with corn flour, bran, and soybean meal, then add vegetable oil, animal oil and water, and mix evenly to obtain the mixture;
(4)制粒：采用制粒机将所述混合料制成颗粒，粒径10mm～12mm，长度25mm；(4) Granulation: the mixture is made into granules by a granulator, with a particle diameter of 10 mm to 12 mm and a length of 25 mm;
(5)烘干：将所述颗粒置于70℃～85℃的烘箱内烘干；(5) drying: drying the particles in an oven at 70°C to 85°C;
(6)灭菌：将烘干颗粒以一定重量分装后采用20kGy 60Co-γ射线照射灭菌；(6) Sterilization: sterilize by irradiating 20kGy 60Co-γ rays after the dried granules are subpackaged by a certain weight;
(7)成品饲料的检测：将步骤(6)制得的成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合国家标准要求。(7) Detection of finished feed: the finished product prepared in step (6) is tested for nutritional components and health indicators, so as to ensure that the indicators of the finished feed meet the requirements of national standards.
实施例2Example 2
本发明Omega3过表达小鼠饲料组合物的一种实施例，本实施例所述Omega3过表达小鼠饲料组合物包含以下重量份的组分：玉米粉31.2份、麸皮7.8份、豆粕9.5份、酪蛋白5.5份、糖类10.5份、赖氨酸11.5份、蛋氨酸5.4份、胱氨酸2.8份、植物油3.8份、动物油脂3.5份、混合矿物质3.5份、混合维生素1.4份、纤维素4.8份。An embodiment of the Omega3 overexpression mouse feed composition of the present invention, the Omega3 overexpression mouse feed composition described in this embodiment comprises the following components by weight: 31.2 parts of corn flour, 7.8 parts of bran, and 9.5 parts of soybean meal , 5.5 parts of casein, 10.5 parts of sugar, 11.5 parts of lysine, 5.4 parts of methionine, 2.8 parts of cystine, 3.8 parts of vegetable oil, 3.5 parts of animal fat, 3.5 parts of mixed minerals, 1.4 parts of mixed vitamins, 4.8 parts of cellulose share.
本实施例中，所述糖类为乳糖；所述植物油为橄榄油和花生油；所述动物油脂为牛油；所述混合矿物质包括碳酸氢钙、氯化钠、硫酸铁、硫酸锌、碘化钾及硫酸铜；所述混合维生素包括维生素A、维生素D、维生素E、维生素K、维生素B1、维生素B2、维生素B6、维生素B12、盐酸、泛酸钙、叶酸及生物素；所述纤维素为聚乙烯纤维素。In this embodiment, the saccharide is lactose; the vegetable oil is olive oil and peanut oil; the animal fat is tallow; the mixed minerals include calcium bicarbonate, sodium chloride, iron sulfate, zinc sulfate, potassium iodide and copper sulfate; the mixed vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B 1 , vitamin B 2 , vitamin B 6 , vitamin B 12 , hydrochloric acid, calcium pantothenate, folic acid and biotin; the fiber The element is polyethylene cellulose.
本实施例所述Omega3过表达小鼠饲料组合物采用如下方法制备而成：The Omega3 overexpression mouse feed composition described in this example is prepared by the following method:
(1)粉碎：将玉米、麸皮、豆粕分别投到粉碎机粉碎，过80～100目，备用；(1) Pulverization: Put corn, bran and soybean meal into a pulverizer for pulverization, pass through 80-100 meshes, and set aside;
(2)预混：将酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、混合矿物质、混合维生素及纤维素进行预混，获得预混料；(2) Premixing: premixing casein, sugars, lysine, methionine, cystine, mixed minerals, mixed vitamins and cellulose to obtain a premix;
(3)混料：将上述预混料与玉米粉、麸皮、豆粕混合均匀，再加入植物油、动物油以及水，混合均匀，获得混合料；(3) Mixing: Mix the above-mentioned premix with corn flour, bran, and soybean meal, then add vegetable oil, animal oil and water, and mix evenly to obtain the mixture;
(4)制粒：采用制粒机将所述混合料制成颗粒，粒径10mm～12mm，长度25mm；(4) Granulation: the mixture is made into granules by a granulator, with a particle diameter of 10 mm to 12 mm and a length of 25 mm;
(5)烘干：将所述颗粒置于70℃～85℃的烘箱内烘干；(5) drying: drying the particles in an oven at 70°C to 85°C;
(6)灭菌：将烘干颗粒以一定重量分装后采用20kGy 60Co-γ射线照射灭菌；(6) Sterilization: sterilize by irradiating 20kGy 60Co-γ rays after the dried granules are subpackaged by a certain weight;
(7)成品饲料的检测：将步骤(6)制得的成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合国家标准要求。(7) Detection of finished feed: the finished product prepared in step (6) is tested for nutritional components and health indicators, so as to ensure that the indicators of the finished feed meet the requirements of national standards.
实施例3Example 3
本发明Omega3过表达小鼠饲料组合物的一种实施例，本实施例所述Omega3过表达小鼠饲料组合物包含以下重量份的组分：玉米粉31.4份、麸皮7.6份、豆粕9份、酪蛋白5份、糖类10份、赖氨酸12份、蛋氨酸5.6份、胱氨酸2.4份、植物油3.4份、动物油脂2.6份、混合矿物质5份、混合维生素1.2份、纤维素4份。An embodiment of the Omega3 overexpression mouse feed composition of the present invention, the Omega3 overexpression mouse feed composition described in this embodiment comprises the following components by weight: 31.4 parts of corn flour, 7.6 parts of bran, and 9 parts of soybean meal , 5 parts of casein, 10 parts of sugar, 12 parts of lysine, 5.6 parts of methionine, 2.4 parts of cystine, 3.4 parts of vegetable oil, 2.6 parts of animal fat, 5 parts of mixed minerals, 1.2 parts of mixed vitamins, 4 parts of cellulose share.
本实施例中，所述糖类为乳糖；所述植物油为玉米油；所述动物油脂为猪油；所述混合矿物质包括碳酸氢钙、氯化钠、硫酸铁、硫酸锌、碘化钾及硫酸铜；所述混合维生素包括维生素A、维生素D、维生素E、维生素K、维生素B1、维生素B2、维生素B6、维生素B12、盐酸、泛酸钙、叶酸及生物素；所述纤维素为木质素。In this embodiment, the saccharide is lactose; the vegetable oil is corn oil; the animal fat is lard; the mixed minerals include calcium bicarbonate, sodium chloride, iron sulfate, zinc sulfate, potassium iodide and sulfuric acid Copper; the mixed vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B 1 , vitamin B 2 , vitamin B 6 , vitamin B 12 , hydrochloric acid, calcium pantothenate, folic acid and biotin; the cellulose is Lignin.
本实施例所述Omega3过表达小鼠饲料组合物采用如下方法制备而成：The Omega3 overexpression mouse feed composition described in this example is prepared by the following method:
(1)粉碎：将玉米、麸皮、豆粕分别投到粉碎机粉碎，过80～100目，备用；(1) Pulverization: Put corn, bran and soybean meal into a pulverizer for pulverization, pass through 80-100 meshes, and set aside;
(2)预混：将酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、混合矿物质、混合维生素及纤维素进行预混，获得预混料；(2) Premixing: premixing casein, sugars, lysine, methionine, cystine, mixed minerals, mixed vitamins and cellulose to obtain a premix;
(3)混料：将上述预混料与玉米粉、麸皮、豆粕混合均匀，再加入植物油、动物油以及水，混合均匀，获得混合料；(3) Mixing: Mix the above-mentioned premix with corn flour, bran, and soybean meal, then add vegetable oil, animal oil and water, and mix evenly to obtain the mixture;
(4)制粒：采用制粒机将所述混合料制成颗粒，粒径10mm～12mm，长度25mm；(4) Granulation: the mixture is made into granules by a granulator, with a particle diameter of 10 mm to 12 mm and a length of 25 mm;
(5)烘干：将所述颗粒置于70℃～85℃的烘箱内烘干；(5) drying: drying the particles in an oven at 70°C to 85°C;
(6)灭菌：将烘干颗粒以一定重量分装后采用20kGy 60Co-γ射线照射灭菌；(6) Sterilization: sterilize by irradiating 20kGy 60Co-γ rays after the dried granules are subpackaged by a certain weight;
(7)成品饲料的检测：将步骤(6)制得的成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合国家标准要求。(7) Detection of finished feed: the finished product prepared in step (6) is tested for nutritional components and health indicators, so as to ensure that the indicators of the finished feed meet the requirements of national standards.
实施例4Example 4
本发明Omega3过表达小鼠饲料组合物的一种实施例，本实施例所述Omega3过表达小鼠饲料组合物包含以下重量份的组分：玉米粉31.5份、麸皮7.4份、豆粕10份、酪蛋白4.5份、糖类9份、赖氨酸12.5份、蛋氨酸6份、胱氨酸2.4份、植物油4份、动物油脂3份、混合矿物质4份、混合维生素1份、纤维素4.6份。An embodiment of the Omega3 overexpression mouse feed composition of the present invention, the Omega3 overexpression mouse feed composition described in this embodiment comprises the following components by weight: 31.5 parts of corn flour, 7.4 parts of bran, and 10 parts of soybean meal , 4.5 parts of casein, 9 parts of sugar, 12.5 parts of lysine, 6 parts of methionine, 2.4 parts of cystine, 4 parts of vegetable oil, 3 parts of animal fat, 4 parts of mixed minerals, 1 part of mixed vitamins, 4.6 parts of cellulose share.
本实施例中，所述糖类为乳糖；所述植物油为菜籽油和亚麻油；所述动物油脂为猪油和牛油；所述混合矿物质包括碳酸氢钙、氯化钠、硫酸铁、硫酸锌、碘化钾及硫酸铜；所述混合维生素包括维生素A、维生素D、维生素E、维生素K、维生素B1、维生素B2、维生素B6、维生素B12、盐酸、泛酸钙、叶酸及生物素；所述纤维素为木质素和羧甲基纤维素。In this embodiment, the saccharide is lactose; the vegetable oil is rapeseed oil and linseed oil; the animal fat is lard and tallow; the mixed minerals include calcium bicarbonate, sodium chloride, iron sulfate, Zinc sulfate, potassium iodide and copper sulfate; the mixed vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B 1 , vitamin B 2 , vitamin B 6 , vitamin B 12 , hydrochloric acid, calcium pantothenate, folic acid and biotin ; The cellulose is lignin and carboxymethyl cellulose.
本实施例所述Omega3过表达小鼠饲料组合物采用如下方法制备而成：The Omega3 overexpression mouse feed composition described in this example is prepared by the following method:
(1)预混：将酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、混合矿物质、混合维生素及纤维素进行预混，获得预混料；(1) Premixing: premixing casein, sugars, lysine, methionine, cystine, mixed minerals, mixed vitamins and cellulose to obtain a premix;
(2)混料：将上述预混料与玉米粉、麸皮、豆粕混合均匀，再加入植物油、动物油以及水，混合均匀，获得混合料；(2) Mixing: Mix the above-mentioned premix with corn flour, bran, and soybean meal, then add vegetable oil, animal oil and water, and mix evenly to obtain the mixture;
(3)制粒：采用制粒机将所述混合料制成颗粒，粒径10mm～12mm，长度25mm；(3) Granulation: the mixture is made into granules by a granulator, with a particle diameter of 10 mm to 12 mm and a length of 25 mm;
(4)烘干：将所述颗粒置于70℃～85℃的烘箱内烘干；(4) drying: drying the particles in an oven at 70°C to 85°C;
(6)灭菌：将烘干颗粒以一定重量分装后采用20kGy 60Co-γ射线照射灭菌；(6) Sterilization: sterilize by irradiating 20kGy 60Co-γ rays after the dried granules are subpackaged by a certain weight;
(7)成品饲料的检测：将步骤(6)制得的成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合国家标准要求。(7) Detection of finished feed: the finished product prepared in step (6) is tested for nutritional components and health indicators, so as to ensure that the indicators of the finished feed meet the requirements of national standards.
实施例5Example 5
本发明Omega3过表达小鼠饲料组合物的一种实施例，本实施例所述Omega3过表达小鼠饲料组合物包含以下重量份的组分：玉米粉31.8份、麸皮7.2份、豆粕8.5份、酪蛋白4份、糖类8.5份、赖氨酸13份、蛋氨酸5.8份、胱氨酸2.2份、植物油4.2份、动物油脂2.6份、混合矿物质5份、混合维生素0.9份、纤维素4.2份。An embodiment of the Omega3 overexpression mouse feed composition of the present invention, the Omega3 overexpression mouse feed composition described in this embodiment comprises the following components by weight: 31.8 parts of corn flour, 7.2 parts of bran, and 8.5 parts of soybean meal , 4 parts of casein, 8.5 parts of sugar, 13 parts of lysine, 5.8 parts of methionine, 2.2 parts of cystine, 4.2 parts of vegetable oil, 2.6 parts of animal fat, 5 parts of mixed minerals, 0.9 parts of mixed vitamins, 4.2 parts of cellulose share.
本实施例中，所述糖类为乳糖；所述植物油为花生油和玉米油；所述动物油脂为猪油和牛油；所述混合矿物质包括碳酸氢钙、氯化钠、硫酸铁、硫酸锌、碘化钾及硫酸铜；所述混合维生素包括维生素A、维生素D、维生素E、维生素K、维生素B1、维生素B2、维生素B6、维生素B12、盐酸、泛酸钙、叶酸及生物素；所述纤维素为羧甲基纤维素和聚乙烯纤维素。In this embodiment, the saccharide is lactose; the vegetable oil is peanut oil and corn oil; the animal fat is lard and tallow; the mixed minerals include calcium bicarbonate, sodium chloride, iron sulfate, zinc sulfate , potassium iodide and copper sulfate; the mixed vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B 1 , vitamin B 2 , vitamin B 6 , vitamin B 12 , hydrochloric acid, calcium pantothenate, folic acid and biotin; Described cellulose is carboxymethyl cellulose and polyethylene cellulose.
本实施例所述Omega3过表达小鼠饲料组合物采用如下方法制备而成：The Omega3 overexpression mouse feed composition described in this example is prepared by the following method:
(1)粉碎：将玉米、麸皮、豆粕分别投到粉碎机粉碎，过80～100目，备用；(1) Pulverization: Put corn, bran and soybean meal into a pulverizer for pulverization, pass through 80-100 meshes, and set aside;
(2)预混：将酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、混合矿物质、混合维生素及纤维素进行预混，获得预混料；(2) Premixing: premixing casein, sugars, lysine, methionine, cystine, mixed minerals, mixed vitamins and cellulose to obtain a premix;
(3)混料：将上述预混料与玉米粉、麸皮、豆粕混合均匀，再加入植物油、动物油以及水，混合均匀，获得混合料；(3) Mixing: Mix the above-mentioned premix with corn flour, bran, and soybean meal, then add vegetable oil, animal oil and water, and mix evenly to obtain the mixture;
(4)制粒：采用制粒机将所述混合料制成颗粒，粒径10mm～12mm，长度25mm；(4) Granulation: the mixture is made into granules by a granulator, with a particle diameter of 10 mm to 12 mm and a length of 25 mm;
(5)烘干：将所述颗粒置于70℃～85℃的烘箱内烘干；(5) drying: drying the particles in an oven at 70°C to 85°C;
(6)灭菌：将烘干颗粒以一定重量分装后采用20kGy 60Co-γ射线照射灭菌；(6) Sterilization: sterilize by irradiating 20kGy 60Co-γ rays after the dried granules are subpackaged by a certain weight;
(7)成品饲料的检测：将步骤(6)制得的成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合国家标准要求。(7) Detection of finished feed: the finished product prepared in step (6) is tested for nutritional components and health indicators, so as to ensure that the indicators of the finished feed meet the requirements of national standards.
实施例6Example 6
本发明Omega3过表达小鼠饲料组合物的一种实施例，本实施例所述Omega3过表达小鼠饲料组合物包含以下重量份的组分：玉米粉32份、麸皮7份、豆粕9份、酪蛋白5份、糖类8份、赖氨酸12份、蛋氨酸5.6份、胱氨酸2份、植物油4.5份、动物油脂2份、混合矿物质4.5份、混合维生素0.8份、纤维素4.5份。An embodiment of the Omega3 overexpression mouse feed composition of the present invention, the Omega3 overexpression mouse feed composition described in this embodiment comprises the following components by weight: 32 parts of corn flour, 7 parts of bran, and 9 parts of soybean meal , 5 parts of casein, 8 parts of sugar, 12 parts of lysine, 5.6 parts of methionine, 2 parts of cystine, 4.5 parts of vegetable oil, 2 parts of animal fat, 4.5 parts of mixed minerals, 0.8 parts of mixed vitamins, 4.5 parts of cellulose share.
本实施例中，所述糖类为乳糖；所述植物油为菜籽油；所述动物油脂为牛油；所述混合矿物质包括碳酸氢钙、氯化钠、硫酸铁、硫酸锌、碘化钾及硫酸铜；所述混合维生素包括维生素A、维生素D、维生素E、维生素K、维生素B1、维生素B2、维生素B6、维生素B12、盐酸、泛酸钙、叶酸及生物素；所述纤维素为木质素。In the present embodiment, the saccharide is lactose; the vegetable oil is rapeseed oil; the animal fat is tallow; the mixed minerals include calcium bicarbonate, sodium chloride, iron sulfate, zinc sulfate, potassium iodide and Copper sulfate; the mixed vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B 1 , vitamin B 2 , vitamin B 6 , vitamin B 12 , hydrochloric acid, calcium pantothenate, folic acid and biotin; the cellulose for lignin.
本实施例所述Omega3过表达小鼠饲料组合物采用如下方法制备而成：The Omega3 overexpression mouse feed composition described in this example is prepared by the following method:
(1)粉碎：对粒状或块状原料，如玉米、麸皮、豆粕分别投到粉碎机粉碎，过80～100目，备用；(1) Crushing: Put granular or massive raw materials, such as corn, bran, and soybean meal, into a pulverizer for pulverization, pass through 80 to 100 meshes, and set aside;
(2)预混：将酪蛋白、糖类、赖氨酸、蛋氨酸、胱氨酸、混合矿物质、混合维生素及纤维素进行预混，获得预混料；(2) Premixing: premixing casein, sugars, lysine, methionine, cystine, mixed minerals, mixed vitamins and cellulose to obtain a premix;
(3)混料：将上述预混料与玉米粉、麸皮、豆粕混合均匀，再加入植物油、动物油以及水，混合均匀，获得混合料；优选地，固体油脂加热完全融化后加入；(3) Mixing material: mix the above premixed material with corn flour, bran, and soybean meal evenly, then add vegetable oil, animal oil and water, and mix evenly to obtain a mixed material; preferably, add the solid oil after heating and melting completely;
(4)制粒：采用制粒机将所述混合料制成颗粒，粒径10mm～12mm，长度25mm；(4) Granulation: the mixture is made into granules by a granulator, with a particle diameter of 10 mm to 12 mm and a length of 25 mm;
(5)烘干：将所述颗粒置于70℃～85℃的烘箱内烘干；(5) drying: drying the particles in an oven at 70°C to 85°C;
(6)灭菌：将烘干颗粒以一定重量分装后采用20kGy 60Co-γ射线照射灭菌；(6) Sterilization: sterilize by irradiating 20kGy 60Co-γ rays after the dried granules are subpackaged by a certain weight;
(7)成品饲料的检测：将步骤(6)制得的成品进行营养成分及卫生指标检测，保证成品饲料的各项指标符合国家标准要求。(7) Detection of finished feed: the finished product prepared in step (6) is tested for nutritional components and health indicators, so as to ensure that the indicators of the finished feed meet the requirements of national standards.
实施例7本发明Omega3过表达小鼠饲料组合物的效果试验Example 7 Effect test of Omega3 overexpression mouse feed composition of the present invention
本实施例主要为本发明Omega3过表达小鼠的饲料组合物与对比例的饲料组合物的性能对比，对比例的饲料组合物采用Omega3过表达小鼠最通用的一种饲料(温和型饲料，含有10％的油脂)。对比例的饲料组合物含有如下重量份的组分：玉米淀粉36.75份、酪蛋白20份、麦芽糖糊精13.2份、蔗糖10份、豆油10份、纤维素5.0份、混合矿物质3.5份、混合维生素1.0份、胱氨酸0.3份、氯化胆碱0.25份。The present embodiment mainly is the performance comparison of the feed composition of the Omega3 overexpression mouse of the present invention and the feed composition of the comparative example, and the feed composition of the comparative example adopts the most general a kind of feed of Omega3 overexpression mouse (mild type feed, Contains 10% fat). The feed composition of the comparative example contains the following components by weight: 36.75 parts of cornstarch, 20 parts of casein, 13.2 parts of maltodextrin, 10 parts of sucrose, 10 parts of soybean oil, 5.0 parts of cellulose, 3.5 parts of mixed minerals, mixed Vitamin 1.0, cystine 0.3, choline chloride 0.25.
1、实验方法1. Experimental method
实验动物分组与饲养管理：56只8周龄SPF级Omega3过表达小鼠随机分为实验对照组(对比例中所述饲料组合物喂养，设为实验对照组)和实验组(本发明所述饲料组合物喂养，设为实验组1、实验组2、实验组3、实验组4、实验组5及实验组6)，每组8只。8只8周龄SPF级C57BL/6J小鼠设为空白对照组(常规标准饲料喂养，设为空白对照组)，试验周期为8周。以上实验动物均由广东省实验动物监测所提供，组织取材在广东省实验动物监测所屏障动物实验设施内进行【SYXK(粤)2012-0122】。动物饲养环境温度为20±2℃，湿度为60±2％，照明时间为12±3h。整个试验过程对动物的饲养及处置符合广东省实验动物饲养设施标准和实验动物3R原则。Experimental animal grouping and feeding management: 56 8-week-old SPF level Omega3 overexpression mice are randomly divided into experimental control group (feed composition described in the comparative example is fed as experimental control group) and experimental group (described in the present invention) Feed composition feeding, set as experimental group 1, experimental group 2, experimental group 3, experimental group 4, experimental group 5 and experimental group 6), each group 8. Eight 8-week-old SPF grade C57BL/6J mice were set as blank control group (fed with conventional standard feed, set as blank control group), and the test period was 8 weeks. The above experimental animals were all provided by the Guangdong Provincial Laboratory Animal Monitoring Institute, and the tissue collection was carried out in the barrier animal experimental facility of the Guangdong Provincial Laboratory Animal Monitoring Institute [SYXK (Guangdong) 2012-0122]. The temperature of the animal breeding environment is 20±2°C, the humidity is 60±2%, and the lighting time is 12±3h. The feeding and disposal of animals during the whole experiment conformed to the standard of experimental animal breeding facilities in Guangdong Province and the 3R principle of experimental animals.
检测方法：将上述小鼠饲喂8周后，1)采用气相色谱法分析心脏组织中脂肪酸含量。取心脏组织30mg，采用甲酯化法提取其脂肪酸后，上机检测。检测结果采用面积归一化法计算心脏组织中某一种脂肪酸含量，分析不同分组小鼠心脏组织中的脂肪酸含量；2)心脏超声影像学检测。应用Vevo2100小动物超高分辨力超声影像系统，采用MS-550D探头，获取清晰的胸骨旁短轴B型图像后，在M型图像下，采集心脏超声心动图参数，评估小鼠心脏形态结构与心功能；3)生物标记物检测。禁食12h后，经眼眶静脉丛采血，静置于4℃冰箱1h后，3000r/min离心10min，取上清液，获取血清，采用全自动生化分析仪检测血清总胆固醇(TC)、总甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、血清肌酸激酶(CK)及乳酸脱氢酶(LDH)含量；此外，小鼠采集血液后安乐死，快速采集心脏，在冰冷的生理盐水中漂洗，除去血液，滤纸拭干，称重，加入适量的冰冷生理盐水，使用组织匀浆仪匀浆心脏(匀浆时间10s/次。间隔30s，连续3～5次，在冰水中进行)，制备的10％匀浆用低温低速离心机3000r/min离心15min，获取上清液，采用luminex液相芯片技术检测小鼠心脏组织中肌钙蛋白(cTnI和cTnT)含量。Detection method: after feeding the above-mentioned mice for 8 weeks, 1) analyze the fatty acid content in heart tissue by gas chromatography. Take 30 mg of heart tissue, use the methyl esterification method to extract its fatty acids, and test it on the machine. The test results use the area normalization method to calculate the content of a certain fatty acid in the heart tissue, and analyze the fatty acid content in the heart tissue of different groups of mice; 2) Cardiac ultrasound imaging detection. Using the Vevo2100 ultra-high-resolution ultrasound imaging system for small animals, the MS-550D probe was used to obtain clear parasternal short-axis B-mode images, and then under the M-mode images, cardiac echocardiographic parameters were collected to evaluate the morphology and structure of the mouse heart. Cardiac function; 3) Biomarker detection. After fasting for 12 hours, blood was collected through the orbital venous plexus, placed in a refrigerator at 4°C for 1 hour, centrifuged at 3000r/min for 10 minutes, and the supernatant was collected to obtain serum, and the serum total cholesterol (TC) and total glycerol were detected by an automatic biochemical analyzer Triesters (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels; in addition, mice collected After the blood was euthanized, the heart was quickly collected, rinsed in ice-cold normal saline, the blood was removed, dried with filter paper, weighed, an appropriate amount of ice-cold normal saline was added, and the heart was homogenized using a tissue homogenizer (homogenization time 10s/time. 30s, 3 to 5 times in a row, carried out in ice water), the prepared 10% homogenate was centrifuged with a low-temperature low-speed centrifuge at 3000r/min for 15min, and the supernatant was obtained, and luminex liquid chip technology was used to detect muscle calcium in mouse heart tissue Protein (cTnI and cTnT) content.
2、统计分析方法2. Statistical analysis methods
所有实验数据使用GraphPad Prism 5软件进行统计分析，所有数据用 表示。组间比较采用t检验，p<0.05表示有显著性差异，p<0.01表示有极显著性差异。All experimental data were statistically analyzed using GraphPad Prism 5 software, and all data were analyzed with express. The t test was used for comparison between groups, p<0.05 indicated significant difference, and p<0.01 indicated extremely significant difference.
3、结果分析3. Result analysis
通过气相色谱仪检测各组小鼠心脏组织中多不饱和脂肪酸的含量，结果见表1。与空白对照组比较，实验对照组与实验组(1-5)小鼠心脏组织中DHA含量显著高于空白对照组(p<0.05)，实验对照组和实验组(1-3/5-6)小鼠心脏组织中LA含量明显降低(p<0.05orp<0.01)，实验组(1-3/5-6)小鼠心脏组织中AA含量显著下降(p<0.05)。此外，实验对照组和实验组(1-6)小鼠心脏组织中ω-3/ω-6PUFAs比值显著提高(p<0.05)。而实验对照组和实验组(1-6)小鼠心脏组织中EPA、DHA、DPA、LA、AA及ω-3/ω-6PUFAs比值均无显著差异(p>0.05)。以上结果表明，本发明的饲料组合物均能保持Omega3过表达小鼠的典型模型特征。The content of polyunsaturated fatty acids in the heart tissue of mice in each group was detected by gas chromatography, and the results are shown in Table 1. Compared with the blank control group, the DHA content in the heart tissue of the experimental control group and the experimental group (1-5) was significantly higher than that of the blank control group (p<0.05), and the experimental control group and the experimental group (1-3/5-6 ) LA content in the heart tissue of the mice was significantly decreased (p<0.05orp<0.01), and the AA content in the heart tissue of the experimental group (1-3/5-6) mice was significantly decreased (p<0.05). In addition, the ratio of ω-3/ω-6 PUFAs in the heart tissue of mice in the experimental control group and experimental group (1-6) was significantly increased (p<0.05). However, there was no significant difference in the ratio of EPA, DHA, DPA, LA, AA and ω-3/ω-6 PUFAs between the experimental control group and the experimental group (1-6) in the heart tissue (p>0.05). The above results show that the feed composition of the present invention can maintain the typical model characteristics of Omega3 overexpression mice.
表1各组小鼠心脏组织中脂肪酸含量(％) Table 1 Fatty acid content (%) in heart tissue of mice in each group
注：LA：亚麻酸，AA：花生四烯酸，EPA：二十碳五烯酸，DPA：二十二碳五烯酸；DHA：二十二碳六烯酸。与空白对照组比较，**表示p<0.01；*表示p<0.05。Note: LA: linolenic acid, AA: arachidonic acid, EPA: eicosapentaenoic acid, DPA: docosapentaenoic acid; DHA: docosahexaenoic acid. Compared with the blank control group, ** indicates p<0.01; * indicates p<0.05.
由图1与图2可知，实验对照组小鼠左心室收缩/舒张末期内径(LVID；s和LVID；d)明显大于空白对照组(p<0.05)，而实验组(1-6)小鼠左心室内径明显减小(p<0.05)，与空白对照组相似。射血分数(EF)与短轴缩短率(FS)是评价心功能的主要指标，其中，EF是反应左心室心肌纤维缩短功能，FS是反应应力-缩短的关系。由图3可知，实验对照组小鼠EF与FS小于空白对照组，而实验组(1-6)小鼠心脏功能明显改善，EF和FS均高于试验对照组，且组间无差异。以上结果表明，Omega3过表达小鼠给予对比例饲料组合物饲喂后心脏形态结构与功能发生改变，而给予本发明饲料组合物饲喂均可明显改善此现象，维持小鼠心脏形态结构与心功能的正常。It can be seen from Figure 1 and Figure 2 that the left ventricular systolic/diastolic end diameter (LVID; s and LVID; d) of the experimental control group mice was significantly larger than that of the blank control group (p<0.05), while the experimental group (1-6) mice The inner diameter of the left ventricle was significantly reduced (p<0.05), which was similar to that of the blank control group. Ejection fraction (EF) and short-axis shortening (FS) are the main indicators for evaluating cardiac function. Among them, EF reflects the shortening function of left ventricular myocardial fibers, and FS reflects the relationship between stress and shortening. It can be seen from Figure 3 that the EF and FS of the mice in the experimental control group were smaller than those in the blank control group, while the cardiac function of the mice in the experimental group (1-6) was significantly improved, and both EF and FS were higher than those in the experimental control group, and there was no difference between the groups. The above results show that the Omega3 overexpression mice are given the changes in heart morphology and function after being fed with the feed composition of the comparison ratio, and feeding with the feed composition of the present invention can significantly improve this phenomenon, and maintain the heart shape and structure of the mice. function normally.
由表2可知，各组小鼠血清TG、TC、LDL-C及HDL-C水平无明显差异(p>0.05)，不具有统计学意义。但作为辅助诊断心肌损伤的生化标记物CK和LDH，在实验对照组小鼠血清中其含量显著高于空白对照组(p<0.05)，而实验组(1-6)小鼠血清中其含量显著低于实验对照组(p<0.05)，且接近空白对照组水平，见图4。同时，肌钙蛋白T(cTnT)和肌钙蛋白(cTnI)具有极高的心肌特异性，目前肌钙蛋白已成为心肌损伤最特异性、最灵敏的心脏分子标志物之一。由图5可知，实验对照组小鼠心脏组织中肌钙蛋白T(cTnT)和肌钙蛋白(cTnI)含量明显低于空白对照组，而实验组(1-6)小鼠心脏组织中其含量明显升高，且与空白对照组数值相近。以上结果均表明，Omega3过表达小鼠给予对比例饲料组合物饲喂后心脏分子标记物出现显著改变，提示心脏可能出现一定程度的损伤，而给予本发明饲料组合物饲喂后心脏分子标记物未见明显改变，提示心脏未出现异常，该结果与超声结果相对应，更进一步证明本发明饲料组合物对小鼠自身机体与心脏无副作用，明显优于对比例饲料组合物。It can be seen from Table 2 that there was no significant difference in serum TG, TC, LDL-C and HDL-C levels of mice in each group (p>0.05), which was not statistically significant. However, as biochemical markers CK and LDH for the auxiliary diagnosis of myocardial injury, their levels in the serum of mice in the experimental control group were significantly higher than those in the blank control group (p<0.05), while their levels in the serum of mice in the experimental group (1-6) Significantly lower than the experimental control group (p<0.05), and close to the level of the blank control group, see Figure 4. At the same time, troponin T (cTnT) and troponin (cTnI) have extremely high myocardial specificity, and troponin has become one of the most specific and sensitive cardiac molecular markers for myocardial injury. It can be seen from Figure 5 that the contents of troponin T (cTnT) and troponin (cTnI) in the heart tissue of the mice in the experimental control group were significantly lower than those in the blank control group, while the contents in the heart tissue of the mice in the experimental group (1-6) were significantly lower than those in the blank control group. Significantly increased, and similar to the value of the blank control group. The above results all show that Omega3 overexpression mice have significant changes in heart molecular markers after being fed with the feed composition of the comparison ratio, suggesting that the heart may be damaged to a certain extent, while giving the feed composition of the present invention After feeding with heart molecular markers There was no obvious change, suggesting that there was no abnormality in the heart. This result corresponds to the ultrasound result, which further proves that the feed composition of the present invention has no side effects on the mouse body and heart, and is obviously better than the feed composition of the comparative example.
表2各组小鼠血清TC、TG、HDL-C和LDL-C水平 Table 2 Serum TC, TG, HDL-C and LDL-C levels of mice in each group
综合以上结果表明，本发明所述的Omega3过表达小鼠饲料组合物既能维持Omega3过表达小鼠典型模型特征，又能够明显改善或消除温和型饲料导致Omega3过表达小鼠心脏形态结构和心功能的损伤，尤其是可以改善心腔扩大的形态学改变。The above results show that the Omega3 overexpression mouse feed composition of the present invention can not only maintain the typical model characteristics of Omega3 overexpression mice, but also can significantly improve or eliminate the heart morphology and cardiac structure of Omega3 overexpression mice caused by mild diet. Impairment of function, especially morphological changes that can improve chamber enlargement.
最后所应当说明的是，以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制，尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that The technical solution of the present invention can be modified or equivalently replaced without departing from the spirit and scope of the technical solution of the present invention.
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