Nutritional composition, food and medicine comprising the same, and uses thereof

Abstract

The present invention relates to a nutritional composition, foods and pharmaceuticals comprising the same, and uses thereof. The nutritional composition comprises medium- and long-chain triglycerides and human milk oligosaccharides, and the weight ratio of the medium- and long-chain triglycerides to the human milk oligosaccharides is from 1:1 to 10:1. The nutritional composition of the present invention can play a synergistic role in improving diarrhea.

Description

Technical Field

[0001] The present invention relates to the field of food, and particularly to a nutritional composition, foods and pharmaceuticals comprising the nutritional composition, and their uses. Background Art

[0002] Diarrhea is a common symptom of the digestive system and one of the main causes of childhood malnutrition, especially in low-income countries or regions. Studies have shown that children under 3 years old experience diarrhea an average of 3 times a year, and each episode of diarrhea deprives children of the nutrients needed for growth. Although diarrhea may seem common, its potential hazards are extensive and severe.

[0003] The causes of diarrhea are diverse, including infectious and non-infectious factors. Among them, infectious factors include viral infections, such as those caused by viruses (e.g., rotavirus), bacteria (e.g., Escherichia coli), parasites, or fungal infections. These pathogens can directly damage the intestinal mucosa or secrete toxins to interfere with normal intestinal function; bacterial infections, such as those caused by Salmonella, Escherichia coli, etc., which cause food poisoning through the ingestion of contaminated food or water, leading to diarrhea; parasitic infections, such as cryptosporidiosis, amoebic dysentery, etc., which are usually transmitted through the ingestion or use of contaminated water. Non-infectious factors mainly include lactose intolerance, that is, due to the lack of sufficient lactase in the body, lactose in dairy products cannot be effectively decomposed, resulting in the fermentation of undigested lactose in the intestine, producing gas and acid, irritating the intestine, and causing diarrhea; drug side effects, such as certain drugs may cause diarrhea; functional gastrointestinal diseases, such as irritable bowel syndrome, inflammatory bowel disease (such as Crohn's disease or ulcerative colitis), etc.

[0004] Lactose intolerance is an important non-infectious factor causing diarrhea. Lactose is a disaccharide whose molecule is composed of glucose and galactose. Lactose cannot be directly absorbed in the human body and needs to be decomposed by lactase before it can be absorbed. However, some people have a lack or deficiency of lactase. After ingesting lactose, lactose cannot be digested and absorbed, and the resulting reaction is called lactose intolerance, that is, the situation of lactose intolerance. It is mainly manifested as symptoms such as abdominal distension, abdominal pain, and diarrhea after eating milk and dairy products; the etiology is related to genetics and small intestinal mucosal lesions; it can occur in any population, but is more common in children.

[0005] Therefore, it is desirable to provide a method that can improve diarrhea, especially diarrhea in infants and / or children. Summary of the Invention

[0006] The present invention is made in view of the above problems existing in the prior art.

[0007] In a first aspect, the present invention relates to a nutritional composition, which comprises:

[0008] Medium-chain and long-chain fatty acid triglycerides; and

[0009] Human milk oligosaccharides,

[0010] wherein the weight ratio of the medium-chain and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 1:1 to 10:1.

[0011] In a second aspect, the present invention relates to a food product comprising the nutritional composition according to the first aspect.

[0012] In a third aspect, the present invention relates to a pharmaceutical product comprising the nutritional composition according to the first aspect.

[0013] In a fourth aspect, the present invention relates to the use of the nutritional composition according to the first aspect or the food product according to the second aspect for non-therapeutic purposes of improving diarrhea.

[0014] In a fifth aspect, the present invention relates to the use of the nutritional composition according to the first aspect in the preparation of a pharmaceutical product for improving diarrhea.

[0015] The inventors unexpectedly found in the research that when medium-chain and long-chain fatty acid triglycerides and human milk oligosaccharides are used in combination according to a certain weight ratio, the two can exert a synergistic effect and significantly synergistically improve diarrhea and lactose intolerance diarrhea. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] To more clearly illustrate the technical solutions of the present invention, the drawings required for describing the embodiments will be briefly described below. It should be understood that these drawings are only for facilitating those skilled in the art to more easily understand the present invention, rather than intending to limit the scope of the present invention.

[0017] Figure 1 Typical graph showing the total opacity of zebrafish intestinal contents after single sample treatment.

[0018] Figure 2 Bar graph showing the total opacity of zebrafish intestinal contents after single sample treatment. Compared with the normal control group: * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0019] In order to make the invention objectives, technical solutions and beneficial technical effects of the present application clearer, the present application will be described in detail below. It should be noted that the various aspects, features, embodiments, and their advantages described in the present application can be compatible and / or combined together.

[0020] Unless otherwise specified, the meanings of the technical terms or scientific and technical terms in this specification are the same as those generally understood by those skilled in the art.

[0021] In this application, unless otherwise specified, the temperature is room temperature (25 °C), the atmosphere is air, and the pressure is atmospheric pressure.

[0022] In this application, the term "medium-chain fatty acid" refers to a fatty acid having 6 to 12 carbon atoms in the carbon chain, such as caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, dodecanoic acid, etc.; the term "long-chain fatty acid" refers to a fatty acid having more than 14 carbon atoms, generally 14 to 30 carbon atoms in the carbon chain, such as myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid, etc. The terms "medium-chain fatty acid triglyceride" and "long-chain fatty acid triglyceride" respectively refer to the esterification reaction products of "medium-chain fatty acid" and "long-chain fatty acid" with glycerol; the term "medium- and long-chain fatty acid triglyceride" refers to triglycerides containing both medium-chain fatty acid residues and long-chain fatty acid residues in the molecular structure.

[0023] In this application, the term "human milk oligosaccharide", also known as "human milk oligosaccharide", is a class of complex oligosaccharides formed by glycosidic bonds connecting structural units such as monosaccharides and derivatives, sialic acid, etc.

[0024] In this application, the term "non-therapeutic purpose" has its conventional meaning in the art, which includes, for example, nutritional and / or health care purposes, such as for the preparation of foods such as ordinary foods, functional foods, health foods or health products, etc.

[0025] The present invention relates to a nutritional composition, foods and drugs comprising the nutritional composition, and their uses.

[0026] The present invention will be described in detail below.

[0027] Nutritional composition

[0028] In a first aspect, the present invention relates to a nutritional composition comprising medium- and long-chain fatty acid triglycerides and human milk oligosaccharides, wherein the weight ratio of the medium- and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 1:1 to 10:1. As an example, the mass ratio of the medium- and long-chain fatty acid triglycerides to the human milk oligosaccharides can be 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, or a range defined by any two of them.

[0029] The inventors unexpectedly found in their research that medium- and long-chain fatty acid triglycerides and human milk oligosaccharides (especially 2'-fucosyllactose, 3'-sialyllactose, and / or lacto-N-neotetraose) can synergistically improve diarrhea, especially lactose intolerance-induced diarrhea, when used in combination in a specific ratio.

[0030] The present invention has no particular requirements for the human milk oligosaccharides used, and the human milk oligosaccharides commonly used in the art can be used. The human milk oligosaccharides can be used in the form of pure products or in the form of non-pure products rich in human milk oligosaccharides.

[0031] Human milk oligosaccharides (also known as milk oligosaccharides or human milk oligosaccharides, Human Milk Oligosaccharides) are the third largest solid component in breast milk, second only to lactose and fat, and have important biological activities. Classified according to representative groups, human milk oligosaccharides can be divided into fucosylated human milk oligosaccharides (such as 2'-fucosyllactose), sialylated human milk oligosaccharides (such as 3'-sialyllactose), and non-fucosylated and non-sialylated human milk oligosaccharides (such as lacto-N-neotetraose).

[0032] In some embodiments, the human milk oligosaccharides are selected from one or more of the following: fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, non-fucosylated and non-sialylated human milk oligosaccharides. For example, the human milk oligosaccharides can be: one or more fucosylated human milk oligosaccharides; one or more sialylated human milk oligosaccharides; one or more non-fucosylated and non-sialylated human milk oligosaccharides; one or more fucosylated human milk oligosaccharides and one or more sialylated human milk oligosaccharides; one or more fucosylated human milk oligosaccharides and one or more non-fucosylated and non-sialylated human milk oligosaccharides; one or more sialylated human milk oligosaccharides and one or more non-fucosylated and non-sialylated human milk oligosaccharides; one or more fucosylated human milk oligosaccharides, one or more sialylated human milk oligosaccharides, and one or more non-fucosylated and non-sialylated human milk oligosaccharides.

[0033] In some embodiments, the fucosylated human milk oligosaccharides are selected from one or more of the following, for example: 2'-fucosyllactose (2'-FL), 3-fucosyllactose, lacto-N-fucopentaose I, lacto-N-difucosahexose I, lacto-N-difucosahexose II, lacto-bifucosyltetraose.

[0034] In some embodiments, the sialylated human milk oligosaccharides are selected from one or more of the following, for example: 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL).

[0035] In some embodiments, the non-fucosylated and non-sialylated human milk oligosaccharides are selected from one or more of the following, for example: lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-triose II, and lacto-N-disaccharide.

[0036] In some embodiments, the human milk oligosaccharide is selected from, for example, one of the following: 2'-fucosyllactose; 3'-sialyllactose; lacto-N-neotetraose; 2'-fucosyllactose and 3'-sialyllactose; 2'-fucosyllactose and lacto-N-neotetraose; 3'-sialyllactose and lacto-N-neotetraose; 2'-fucosyllactose, 3'-sialyllactose and lacto-N-neotetraose.

[0037] In some embodiments, the human milk oligosaccharide is selected from, for example, one of the following: 2'-fucosyllactose; lacto-N-neotetraose; 2'-fucosyllactose and lacto-N-neotetraose.

[0038] When the human milk oligosaccharide is a mixture of any two or more components, the ratio between the components is not particularly limited and can be appropriately selected in consideration of the other effects of each specific human milk oligosaccharide; for example, the mass ratio between any two specific human milk oligosaccharides can be between 20:1 and 1:20, for example, 15:1 to 1:15.

[0039] For example, when the human milk oligosaccharide includes fucosylated human milk oligosaccharide (e.g., 2'-FL) and sialylated human milk oligosaccharide (e.g., 3'-SL), the mass ratio of fucosylated human milk oligosaccharide / sialylated human milk oligosaccharide, for example, the 2'-FL / 3'-SL mass ratio, can be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them.

[0040] For example, when the human milk oligosaccharides include fucosylated human milk oligosaccharides (e.g., 2’-FL) and non-fucosylated and non-sialylated human milk oligosaccharides (e.g., LNnT), the mass ratio of fucosylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 2’-FL / LNnT mass ratio, can be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them.

[0041] For example, when the human milk oligosaccharides include sialylated human milk oligosaccharides (e.g., 3’-SL) and non-fucosylated and non-sialylated human milk oligosaccharides (e.g., LNnT), the mass ratio of sialylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 3’-SL / LNnT mass ratio, can be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them.

[0042] For example, when the human milk oligosaccharides include fucosylated human milk oligosaccharides (such as 2’-FL), sialylated human milk oligosaccharides (such as 3’-SL), and non-fucosylated and non-sialylated human milk oligosaccharides (such as LNnT), the mass ratio of fucosylated human milk oligosaccharides to sialylated human milk oligosaccharides, such as the 2’-FL / 3’-SL mass ratio, the mass ratio of fucosylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 2’-FL / LNnT mass ratio, and the mass ratio of sialylated human milk oligosaccharides to non-fucosylated and non-sialylated human milk oligosaccharides, such as the 3’-SL / LNnT mass ratio, can each independently be 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, or a range defined by any two of them. For example, when the human milk oligosaccharides are a three-component combination of fucosylated human milk oligosaccharides, sialylated human milk oligosaccharides, and non-fucosylated and non-sialylated human milk oligosaccharides, based on 4 parts by weight of non-fucosylated and non-sialylated human milk oligosaccharides (such as LNnT), the amount range of fucosylated human milk oligosaccharides (such as 2’-FL) is 1 to 20, preferably 8 to 28 parts by weight, and / or the amount range of sialylated human milk oligosaccharides (such as 3’-SL) is 1 to 8, preferably 1 to 4 parts by weight.

[0043] Medium-chain triglycerides (MLCT) are a type of structured lipid in which the triglyceride molecule contains both medium-chain fatty acids and long-chain fatty acids, and they have different physical and chemical properties, metabolic characteristics, and nutritional values from long-chain triglycerides (LCT) or medium-chain triglycerides (MCT). Research (see, for example, Yuan Tinglan, Composition and Metabolic Characteristics of Medium-chain Triglycerides in Human Milk Fat [D], Jiangnan University, 2021) shows that medium-chain triglycerides (MLCT) are not simply equivalent to a physical mixture of long-chain triglycerides (LCT) and medium-chain triglycerides (MCT) (nor, of course, to a simple mixture of long-chain fatty acids and medium-chain fatty acids), and the latter do not possess the physical and chemical properties, metabolic characteristics, and nutritional values of the former.

[0044] The present invention has no special requirements for the medium and long-chain fatty acid triglycerides used, and the medium and long-chain fatty acid triglycerides commonly used in the art can be used. The medium and long-chain fatty acid triglycerides can be used in the form of pure products or in the form of non-pure products rich in medium and long-chain fatty acid triglycerides. In some embodiments, the medium and long-chain fatty acid triglycerides of the present invention contain C6-C 12 (such as C6, C7, C8, C9, C 10 , C 11 , C 12 , or the range defined by any two of them) fatty acid residues and C 14 -C 30 (such as C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , C 24 , C 25 , C 26 , C 27 , C 28 , C 29 , C 30 , or the range defined by any two of them) fatty acid residues, wherein:

[0045] The C6-C 12 fatty acid residues are derived from one or more of the following: caproic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, etc., preferably caproic acid, octanoic acid, decanoic acid, and lauric acid; and / or

[0046] The C 14 -C 30 fatty acid residues are derived from one or more of the following: myristic acid, palmitic acid, heptadecanoic acid, oleic acid, linoleic acid, stearic acid, nonadecanoic acid, eicosapentaenoic acid, heneicosanoic acid, docosatetraenoic acid, docosahexaenoic acid, tricosanoic acid, tetracosenoic acid, pentacosapentaenoic acid, docosahexaenoic acid, arachidonic acid, etc., preferably myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid.

[0047] In some embodiments, the weight ratio of the C6-C 12 fatty acid residues to the C 14 -C 30 fatty acid residues in the medium and long-chain fatty acid triglycerides is 0.124 to 2.000. As an example, the C6-C 12The fatty acid residue to the C 14 -C 30 The weight ratio of the fatty acid residue can be 0.124, 0.125, 0.130, 0.140, 0.150, 0.160, 0.170, 0.180, 0.190, 0.200, 0.300, 0.400, 0.500, 0.600, 0.700, 0.800, 0.900, 1.000, 1.100, 1.200, 1.300, 1.400, 1.500, 1.600, 1.700, 1.800, 1.900, 2.000, or within the range defined by any two of them.

[0048] In some embodiments, the nutritional composition of the present invention may be composed of medium and long chain fatty acid esters and human milk oligosaccharides.

[0049] Food

[0050] In a second aspect, the present invention relates to a food comprising the nutritional composition according to the first aspect.

[0051] In some embodiments, the food is an infant food such as infant formula.

[0052] In some embodiments, based on the weight of the food, the content of medium and long chain triglycerides can be 0.1 to 30% by weight. As an example, based on the weight of the food, the content of medium and long chain triglycerides can be 0.1, 0.2, 0.5, 0.8, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30% by weight, or within the range defined by any two of them.

[0053] In some embodiments, based on the weight of the food, the content of human milk oligosaccharides can be 0.01 to 6% by weight. As an example, based on the weight of the food, the content of human milk oligosaccharides can be 0.01, 0.02, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6% by weight, or within the range defined by any two of them.

[0054] In some embodiments, the food can be, for example, a functional food, a health food, or a health product.

[0055] In some embodiments, the food is a general food or a special food. As an example, the food can be selected from dairy products, candies, beverages, breads, and biscuits, for example, the food can be selected from milk powder or fermented foods.

[0056] In some embodiments, the milk powder is infant formula milk powder.

[0057] In addition, it is easy for those skilled in the art to understand that in addition to the nutritional composition of the first aspect of the present invention, the food further comprises one or more selected from the following: fresh raw milk, demineralized whey powder, concentrated whey protein, lactose, edible vegetable blend oil, fructooligosaccharide, galactooligosaccharide, nucleotides, choline, vitamins, minerals, DHA, and taurine.

[0058] For example, when the food is milk powder, in addition to the nutritional composition, the milk powder may further comprise proteins such as α-lactalbumin and milk fat globule membrane protein; saccharides such as lactose; lipids; minerals such as calcium, iron, phosphorus, etc.; vitamins; and other additives such as whey powder, choline bitartrate, docosahexaenoic acid, arachidonic acid, walnut oil, etc.

[0059] The food can be produced by the preparation methods commonly used in the art, which are well known to those skilled in the art and will not be elaborated herein.

[0060] All of the above descriptions regarding the nutritional composition of the first aspect of the present invention are applicable here and will not be elaborated herein.

[0061] Medicine

[0062] In a third aspect, the present invention relates to a medicine which comprises the nutritional composition according to the first aspect.

[0063] The present invention has no particular requirement for the dosage form of the medicine, and the commonly used dosage forms in the art can be adopted. As an example, the medicine of the present invention can be solid preparations, such as powders, granules, tablets, capsules, dripping pills, pills, etc.; semi-solid preparations, such as ointments, creams, eye ointments, gels, etc.; liquid preparations, such as oral liquid preparations, such as syrups, mixtures, suspensions, emulsions, etc.

[0064] It is easy for those skilled in the art to understand that according to the differences in the dosage form of the medicine and the application scenarios, etc., the medicine may further comprise various pharmaceutically acceptable excipients. In addition, the medicine of the present invention can be prepared by the methods commonly adopted in the art, which are well known to those skilled in the art and will not be elaborated herein.

[0065] All of the above descriptions regarding the nutritional composition of the first aspect of the present invention are applicable here and will not be elaborated herein.

[0066] Use

[0067] In a fourth aspect, the present invention relates to the use of the nutritional composition according to the first aspect or the food according to the second aspect for promoting bone development for non-therapeutic purposes.

[0068] Said improvement of diarrhea may, for example, include improvement of lactose intolerance diarrhea.

[0069] Said improvement of diarrhea may, for example, include, but is not limited to, improvement of diarrhea in infants and / or children and / or adolescents.

[0070] Said non-therapeutic purposes may, for example, include nutritional and / or health care purposes, such as for preparing foods such as ordinary foods, functional foods, health foods or health products, etc.

[0071] In some embodiments, when the nutritional composition or food is taken orally by a human body, the dosage thereof is such that the dosage of the medium-chain and long-chain fatty acid triglycerides is 0.1 to 30 g / day, preferably 2.5 to 30 g / day. As an example, the dosage of the medium-chain and long-chain fatty acid triglycerides may be 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 g / day, or within the range defined by any two of them.

[0072] In some embodiments, when the nutritional composition or food is taken orally by a human body, the dosage thereof is such that the dosage of the human milk oligosaccharide is 0.05 to 25 g / day, preferably 0.1 to 25 g / day. As an example, the dosage of the human milk oligosaccharide may be 0.05, 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 g / day, or within the range defined by any two of them, and the dosage of the medium-chain and long-chain fatty acid triglycerides can be converted according to the ratio to the human milk oligosaccharide.

[0073] In a fifth aspect, the present invention relates to the use of the nutritional composition according to the first aspect in the preparation of a medicament for improving diarrhea.

[0074] All of the above descriptions of the nutritional composition in the first aspect, the food in the second aspect and the medicament in the third aspect of the present invention are applicable herein and will not be repeated here.

[0075] Examples

[0076] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below in conjunction with examples. It should be understood that the specific examples described herein are only used to explain the present invention and are not used to limit the present invention.

[0077] Experimental principle description

[0078] In this experiment, a lactose-induced diarrhea model of lactose intolerance in zebrafish was used.

[0079] Compared with traditional animal experiments (piglets, rats and mice) and human experiments, the zebrafish experimental protocol has many advantages in the evaluation experiment of the efficacy of promoting growth (bone) development. The zebrafish experimental protocol has many advantages in the evaluation experiment of the efficacy of improving diarrhea. Compared with traditional animal experiments (such as piglets, rats and mice) and human experiments, it is specifically reflected in the following aspects: (1) Fast and efficient: The experimental period of the zebrafish model is shorter, the operation is simple, the results are easy to observe and the cost is low. This makes zebrafish an efficient in vivo animal model, capable of realizing early, rapid and high-throughput screening of drugs for promoting intestinal function activity. (2) High safety: The zebrafish model can effectively improve the screening speed of drugs while ensuring the safety of the screened drugs. This is because the physiology, development and metabolism of zebrafish are highly similar to those of mammalian animals, and the homology of genes with human genes reaches 87%, which can reliably simulate and predict human physiological and pathological processes. (3) High-throughput screening: The zebrafish model can achieve high-throughput screening, which is very beneficial for the early testing and evaluation of candidate drugs in the new drug R & D process. In addition, the application of the zebrafish model can also save a large amount of experimental costs. (4) Similar intestinal composition: The intestinal composition of zebrafish is similar to that of humans, both composed of endothelial cells, connective tissue, circular muscle and outer longitudinal muscle, which gives it unique advantages in simulating human intestinal function. (5) Standardized procedures: The first complete zebrafish technical procedures for food health function detection in China have been released and have been recognized and supported by authoritative scientific research institutions and enterprises. These procedures provide scientific and rigorous technical guidance for zebrafish experiments, ensuring the accuracy and repeatability of experimental results. In short, the intestine of zebrafish is highly homologous to the mammalian intestine in development, tissue and function. The gene expression and transcriptional regulation in intestinal epithelial cells are highly conserved with those of mammals, and the metabolic function is also conserved. The digestive system of zebrafish basically functions 5 days after fertilization, and it is an important model organism for studying the basic processes of intestinal inflammation and injury.

[0080] Lactose intolerance (LI) refers to the abdominal and systemic symptoms that occur in people with lactose malabsorption after ingesting lactose, such as abdominal pain, bloating, and diarrhea. A small number of patients may experience headache, nausea, etc. Lactose is one of the short-chain fermentable carbohydrates, and they can cause symptoms through similar mechanisms. LI accounts for 69% of the total population with food intolerance, ranking first. Among them, 56.6% are also combined with multiple food intolerances, seriously affecting the quality of life of patients. The typical symptoms of lactose intolerance include abdominal pain, bloating, flatulence, diarrhea, and less frequently nausea and vomiting. The mechanisms and temporal correlations of systemic symptoms are still controversial. Abdominal pain and bloating are caused by the fermentation of unabsorbed lactose in the colon. The intestinal flora provides a compensatory pathway for lactose digestion by breaking down lactose into short-chain fatty acids and gases. Unabsorbed lactose in the ileum and colon leads to acidification of the colonic contents and an increase in osmotic load. This results in an increased secretion of electrolytes and fluids, as well as a rapid transit time, leading to loose stools and diarrhea. The zebrafish intestine is directly connected to the esophagus and is a compartmentalized tubular structure, which is divided into three regions: the foregut, midgut, and hindgut. The foregut of zebrafish, also known as the bulbus entericus, is larger than the mid- and hindgut parts and can act as a storage organ equivalent to the stomach. The intestine of zebrafish is highly homologous to the mammalian intestine in terms of development, tissue, and function. The gene expression and transcriptional regulation in intestinal epithelial cells are highly conserved with mammals, and the metabolic functions are also conserved. The digestive system of zebrafish basically functions 5 days after fertilization, and it is an important model organism for studying the basic processes of intestinal inflammation and injury.

[0081] The sources and specifications of the raw materials used in the following examples are as follows:

[0082] Medium-chain and long-chain fatty acid triglycerides: derived from medium-chain and long-chain fatty acid edible oil (provided by Qingdao Haizhiyuan Life Science Co., Ltd., batch number Y1501-23070101), purity 69.8%;

[0083] 2'-Fucosyllactose: provided by DSM Vitamins Trading (Shanghai) Co., Ltd., SHANGHAI, purity 99.2%, batch number: 21107002;

[0084] 3'-Sialyllactose: provided by DSM Vitamins Trading (Shanghai) Co., Ltd., SHANGHAI, purity 95.2%, batch number: DK21294101;

[0085] Lacto-N-neotetraose: provided by DSM Vitamins Trading (Shanghai) Co., Ltd., DTL DutyPaid.Shanghai, purity 100.5%, batch number: 20454001;

[0086] Wild-type AB strain zebrafish: Provided by the fish breeding center of Hangzhou Huante Biotechnology Co., Ltd., and the license number for the use of experimental animals is: SYXK(Zhe)2022-0004.

[0087] Positive control drug

[0088] Galactooligosaccharides.

[0089] In the following examples, AB strain zebrafish were used for animal experiments.

[0090] 1. Detection materials

[0091] 1.1. Sample preparation information

[0092] MLCT, 2’-FL, LNnT and 3’-SL, and the solvent is standard dilution water.

[0093] Positive control: Galactooligosaccharides, white powder, batch number C15916553, Shanghai Macklin Biochemical Co., Ltd., and the solvent is standard dilution water.

[0094] 1.2. Experimental animals

[0095] Zebrafish were all raised in fish culture water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450 - 550 μS / cm; the pH was 6.5 - 8.5; the hardness was 50 - 100 mg / L CaCO3), provided by the fish breeding center of our company, and the license number for the use of experimental animals is: SYXK(Zhe)2022-0004. The feeding management meets the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethical review number: IACUC-2024-10070-01.

[0096] 1.3. Instruments, consumables and reagents

[0097] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); Precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Beilanbo Biotechnology Co., Ltd., China); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China).

[0098] Lactose, anhydrous (batch number D2328302, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Egg yolk powder (batch number 20230203, Zhejiang Aige Biotechnology Co., Ltd., China).

[0099] 2. Detection methods

[0100] 2.1. Determination of the maximum detection concentration (MTC)

[0101] Randomly select wild-type AB strain zebrafish at 4 days post-fertilization (4 dpf) and place them in a 6-well plate, with 30 zebrafish in each well (experimental group). Administer the sample in water (concentrations are shown in Table 1). At the same time, set up a normal control group and a model control group, with a volume of 3 mL per well. After treatment at 28 °C for 1 day, wash away the sample. Except for the normal control group, lactose and egg yolk powder are administered in water to the zebrafish in each concentration group to establish a lactose intolerance model. After treatment at 28 °C for 4 hours, wash away the lactose and egg yolk powder, let it stand for 2 hours, and determine the MTC of the sample for the model zebrafish.

[0102] 2.2. Evaluation of the efficacy of a single sample in improving diarrhea

[0103] Randomly select wild-type AB strain zebrafish at 4 dpf and place them in a 6-well plate, with 30 zebrafish in each well (experimental group). Administer the sample in water (concentrations are shown in Table 2). The positive control is galactooligosaccharide at a concentration of 2000 μg / mL. At the same time, set up a normal control group and a model control group, with a volume of 3 mL per well. After treatment at 28 °C for 1 day, wash away the sample. Except for the normal control group, lactose and egg yolk powder are administered in water to the zebrafish in each concentration group to establish a lactose intolerance model. After treatment at 28 °C for 4 hours, wash away the lactose and egg yolk powder, let it stand for 2 hours. Randomly select 10 zebrafish from each experimental group and place them under a dissection microscope for photographing. Use NIS-Elements D3.20 advanced image processing software to analyze and collect data, and analyze the total opacity of the intestinal contents of the zebrafish. Evaluate the efficacy of the sample in improving lactose intolerance (diarrhea) based on the statistical analysis results of this index. The statistical treatment results are expressed as mean ± SE. Use SPSS 26.0 software for statistical analysis, and p < 0.05 indicates that the difference is statistically significant.

[0104] 2.3. Evaluation of the efficacy of the composition in improving diarrhea

[0105] Randomly select 4 dpf wild-type AB strain zebrafish into 6-well plates, with 30 zebrafish in each well (experimental group). Administer the sample in water solution (concentrations are shown in Table 3), with the positive control being galactooligosaccharides at a concentration of 2000 μg / mL. At the same time, set up a normal control group and a model control group, with a volume of 3 mL per well. After treatment at 28 °C for 1 day, wash away the sample. Except for the normal control group, lactose and egg yolk powder are administered in water solution to zebrafish in each concentration group to establish a lactose intolerance model. After treatment at 28 °C for 4 hours, wash away lactose and egg yolk powder, and let it stand for 2 hours. Randomly select 10 zebrafish from each experimental group and place them under a dissection microscope for photography. Use NIS-Elements D3.20 advanced image processing software to analyze and collect data, and analyze the total opacity of zebrafish intestinal contents. Evaluate the efficacy of the sample in improving lactose intolerance (diarrhea) based on the statistical analysis results of this index. The statistical processing results are expressed as mean ± SE. Use SPSS 26.0 software for statistical analysis, and p < 0.05 indicates that the difference is statistically significant.

[0106] 3. Detection results

[0107] 3.1. MTC

[0108] Under the conditions of this experiment, the MTCs of MLCT, 2’-FL, LNnT, and 3’-SL in improving diarrhea efficacy are 2000 μg / mL, 2000 μg / mL, 2000 μg / mL, and 1000 μg / mL respectively. See Table 1 for details.

[0109] Table 1. Experimental results of the maximum detection concentration (MTC) of the sample (n = 30)

[0110]

[0111] From the results in Table 1, it can be seen that under the conditions of this experiment, when the maximum detection concentration (MTC) of MLCT, 2’-FL, and LNnT is 2000 μg / mL, the state of zebrafish is similar to that of the model control group; while when the treatment concentration of 3’-SL is 1500 μg / mL, the state is worse than that of the model control group, indicating intolerance at this concentration, and when it is 1000 μg / mL, the state of zebrafish is similar to that of the model control group. Therefore, the usage concentration of 3’-SL in subsequent experiments does not exceed 1000 μg / mL.

[0112] 3.2. Efficacy of single sample in improving diarrhea

[0113] Under the conditions of this experiment, MLCT, 2’-FL, LNnT, and 3’-SL all have the efficacy of improving diarrhea, specifically manifested as improving diarrhea caused by lactose intolerance. See Table 2 for details. Figure 1 and Figure 2 。

[0114] Table 2. Experimental results of the improvement of diarrhea by single sample (n = 10)

[0115]

[0116] Compared with the model control group, **p < 0.01, ***p < 0.001

[0117] Figure 1 The typical graph of the total opacity of the intestinal contents of zebrafish after single sample treatment is shown. It can be seen from the figure that in the area marked by the dotted line, compared with the normal control group, the samples treated with 2’-FL, LNnT, MLCT, and 3’-SL all showed a significant increase in the total opacity of the intestinal contents, confirming the effect of improving diarrhea.

[0118] Figure 2 The bar graph of the total opacity of the intestinal contents of zebrafish after single sample treatment is shown. This figure is based on the data in Table 2.

[0119] 3.3. Efficacy of the composition in improving diarrhea

[0120] The measurement results of the total opacity of the intestinal contents of zebrafish by the composition are shown in Table 3 below.

[0121] Table 3. Total opacity of intestinal contents obtained by compositions with different ratios

[0122]

[0123] Note: The amounts of each formula in the table are expressed as follows: MLCT + 2’-FL + LNnT + 3’-SL (μg / mL) = total amount, the same below.

[0124] Table 3 (continued). Total opacity of intestinal contents obtained by compositions with different ratios

[0125]

[0126] Table 3 (continued). Total opacity of intestinal contents obtained by compositions with different ratios

[0127]

[0128] Table 3 (continued). Total opacity of intestinal contents obtained by compositions with different ratios

[0129]

[0130] 3.3.1. Analysis of experimental results

[0131] For the convenience of result analysis and elaboration, the results in Table 3 are further split and combined into Tables 4 - 11 respectively.

[0132] (1) MLCT + a type of human milk oligosaccharide

[0133] The effects of MLCT and a type of human milk oligosaccharide on the total opacity (pixels) of zebrafish intestinal contents are summarized in Tables 4 - 7 below.

[0134] Table 4. Effects of MLCT + 2’-FL on the total opacity of zebrafish intestinal contents

[0135]

[0136] Based on the results in Table 4, it can be seen that compared with MLCT alone (formulation 1) and 2’-FL alone (formulation 2), the combinations of MLCT and 2’-FL with a ratio of MLCT to 2’-FL from 1:1 to 9:1 (formulations 18, 19, and 7) achieved a significant increase in the total opacity of intestinal contents, indicating that the above combinations of MLCT and 2’-FL have a synergistic effect in improving diarrhea.

[0137] Table 5. Effects of MLCT + LNnT on the total opacity of zebrafish intestinal contents (total concentration 400 μg / mL)

[0138]

[0139] Based on the results in Table 5, it can be seen that compared with MLCT alone (formulation 1), LNnT alone (formulation 16), and the combination of MLCT and LNnT with a ratio of MLCT to LNnT of 0.1:1 (formulation 14), the combinations of MLCT and LNnT with a ratio of MLCT to LNnT from 1:1 to 9:1 (formulations 6, 21, and 20) achieved a significant increase in the total opacity of intestinal contents, indicating that the combinations of MLCT and LNnT in formulations 6, 21, and 20 have a synergistic effect in improving diarrhea.

[0140] Table 6. Effects of MLCT + LNnT on the total opacity of zebrafish intestinal contents (total concentration 1000 μg / mL)

[0141]

[0142] Based on the results in Table 6, it can be seen that compared with MLCT alone (formulation 28) and LNnT alone (formulation 25), the combinations of MLCT and LNnT within the range of a ratio of MLCT to LNnT from 1:1 to 9:1 (formulations 24, 26, and 27) achieved a significant increase in the total opacity of intestinal contents, indicating that the combination of MLCT and LNnT has a synergistic effect in improving diarrhea.

[0143] Therefore, as can be seen from Table 5-6, at different dosages, the combination of MLCT and LNnT has a synergistic effect in improving diarrhea.

[0144] Table 7. Effects of MLCT + 3'-SL on the total opacity of zebrafish intestinal contents

[0145]

[0146] According to the results in Table 7, it can be seen that compared with MLCT alone (formulation 1) and 3'-SL alone (formulation 17), the combinations of MLCT and 3'-SL with a ratio of 1:1 to 9:1 (formulations 23, 5, and 22) achieved a significant increase in the total opacity of intestinal contents, indicating a synergistic effect of this combination of MLCT and 3'-SL in improving diarrhea.

[0147] (2) MLCT + two types of human milk oligosaccharides

[0148] The effects of MLCT and two types of human milk oligosaccharides on the total opacity of zebrafish intestinal contents (pixels) are summarized in Tables 8-10 below.

[0149] Table 8. Effects of MLCT + (2'-FL + LNnT) on the total opacity of zebrafish intestinal contents

[0150]

[0151] According to the results in Table 8, it can be seen that compared with MLCT alone (formulation 1), 2'-FL alone (formulation 2), LNnT alone (formulation 16), and (2'-FL + LNnT) alone (formulation 3), the combination of MLCT and (2'-FL + LNnT) with a ratio of 5:1 (formulation 8) achieved a significant increase in the total opacity of intestinal contents. This indicates a synergistic effect of the above combination of MLCT and (2'-FL + LNnT) in improving diarrhea.

[0152] Table 9. Effects of MLCT + (LNnT + 3'-SL) on the total opacity of zebrafish intestinal contents

[0153]

[0154] According to the results in Table 9, it can be seen that compared with MLCT alone (Formulation 1), LNnT alone (Formulation 16), and 3'-SL alone (Formulation 17), the combination of MLCT and (LNnT + 3'-SL) with a ratio of 6:1 (Formulation 9) achieved a significant increase in the total opacity of intestinal contents. This indicates that there is a synergistic effect in improving diarrhea for the above combination of MLCT and (LNnT + 3'-SL).

[0155] Table 10. Effects of MLCT + (2'-FL + 3'-SL) on the total opacity of intestinal contents in zebrafish

[0156]

[0157] According to the results in Table 10, it can be seen that compared with MLCT alone (Formulation 1), 2'-FL alone (Formulation 2), and 3'-SL alone (Formulation 17), the combination of MLCT and (2'-FL + 3'-SL) with a ratio of 7:1 (Formulation 10) achieved a significant increase in the total opacity of intestinal contents. This indicates that there is a synergistic effect in improving diarrhea for the above combination of MLCT and (2'-FL + 3'-SL).

[0158] (3) MLCT + three types of human milk oligosaccharides

[0159] The effects of MLCT and three types of human milk oligosaccharides on the total opacity (pixels) of intestinal contents in zebrafish are summarized in Table 11 below.

[0160] Table 11. Effects of MLCT + (2'-FL + LNnT + 3'-SL) on the total opacity of intestinal contents in zebrafish

[0161]

[0162] According to the results in Table 11, it can be seen that compared with MLCT alone (Formulation 1), 2'-FL alone (Formulation 2), LNnT alone (Formulation 16), and 3'-SL alone (Formulation 17), the combinations of MLCT and (2'-FL + LNnT + 3'-SL) with ratios from 1:1 to 10:1 (Formulation 11 and Formulation 12) achieved a significant increase in the total opacity of intestinal contents. This indicates that there is a synergistic effect in improving diarrhea for the above combinations of MLCT and (2'-FL + LNnT + 3'-SL).

[0163] The above-described are only exemplary embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the inventive concept of the present invention, improvements can still be made to the present invention, but these all fall within the protection scope of the present invention.

Claims

1. A nutritional composition, characterized in that It includes: Medium and long-chain fatty acid triglycerides; and human milk oligosaccharides, wherein the weight ratio of the medium and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 1:1 to 10:1, characterized in that the human milk oligosaccharides are selected from one of the following: 2'-fucosyllactose; 3'-sialyllactose; lacto-N-neotetraose; 2'-fucosyllactose and 3'-sialyllactose; 2'-fucosyllactose and lacto-N-neotetraose; 3'-sialyllactose and lacto-N-neotetraose; 2'-fucosyllactose, 3'-sialyllactose and lacto-N-neotetraose.

2. The nutritional composition according to claim 1, wherein The weight ratio of the medium and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 2:1 to 8:

1.

3. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharides are selected from one of the following: 2'-fucosyllactose; lacto-N-neotetraose; 2'-fucosyllactose and lacto-N-neotetraose.

4. The nutritional composition according to claim 1, characterized in that The human milk oligosaccharides are a combination of 2'-fucosyllactose, 3'-sialyllactose and lacto-N-neotetraose, and the weight ratio of the medium and long-chain fatty acid triglycerides to the human milk oligosaccharides is from 1:1 to 3:

1.

5. A food, which comprises the nutritional composition according to any one of claims 1-4.

6. The food according to claim 5, wherein The food is an infant food.

7. The food according to claim 5, characterized in that The food is selected from dairy products, candies, beverages, breads and biscuits.

8. The food according to claim 5, characterized in that The food is milk powder or a fermented food.

9. The food according to claim 8, characterized in that The milk powder is infant formula milk powder.

10. The food according to any one of claims 5-9, characterized in that The content of the medium and long-chain fatty acid triglycerides is 0.1% to 30%, and the content of the human milk oligosaccharides is 0.01% to 6%, based on the weight of the food.

11. The food according to any one of claims 5-9, which further comprises one or more selected from the following: raw cow milk, demineralized whey powder, concentrated whey protein, lactose, edible vegetable blended oil, fructooligosaccharide, galactooligosaccharide, nucleotides, choline, vitamins, minerals, DHA and taurine.

12. A medicine, which comprises the nutritional composition according to any one of claims 1-4.

13. Use of the nutritional composition according to any one of claims 1-4 or the food according to any one of claims 5-11 for non-therapeutic purposes of improving diarrhea.

14. The use according to claim 13, characterized in that The weight ratio of the medium and long-chain fatty acid triglycerides to the human milk oligosaccharides is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1 or 10:

1.

15. The use according to claim 13, characterized in that The diarrhea includes lactose intolerance diarrhea.

16. The use according to any one of claims 13-15, characterized in that The improvement of diarrhea includes the improvement of diarrhea in infants and / or children.

17. Use of the nutritional composition according to any one of claims 1-4 in the preparation of a medicine for improving diarrhea.

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