Nutritional composition, food containing same and application of nutritional composition

Abstract

The invention relates to a nutritional composition. The nutritional composition comprises medium-chain and long-chain fatty acid triglyceride and nervonic acid in a mass ratio of (3: 1)-(1500: 1), and has a synergistic effect in the aspects of preventing or relieving diarrhea, inhibiting staphylococcus aureus in vivo or in vitro and / or preventing or treating inflammation. The invention also relates to a food product comprising said nutritional composition. The invention also relates to application of the nutritional composition in preparation of products with one or more purposes selected from 1) to 3): 1) preventing or relieving diarrhea; 2) inhibiting staphylococcus aureus in vivo or in vitro; and 3) preventing or treating inflammation.

Description

Technical Field

[0001] The present disclosure relates to the field of food technology, and in particular to a nutritional composition, a food containing the nutritional composition, and applications of the nutritional composition. Background Art

[0002] The latest report in The Lancet shows that Staphylococcus aureus (S. aureus) caused 1.1 million deaths worldwide in 2019, ranking first among 33 major human pathogens (Lakhundi S et al. Clin Microbiol Rev. 2018 Sep 12;31(4):e00020-18). At the same time, S. aureus is also the Gram-positive pathogen with the highest risk of clinical infection in my country. Studies have reported that S. aureus can grow and reproduce in host epithelial cells, endothelial cells, fibroblasts, keratinocytes, and phagocytic cells such as neutrophils and macrophages. Its intracellular survival has been shown to be one of the key factors for S. aureus to evade immune system surveillance and antibiotic killing (Lakhundi S et al. Clin Microbiol Rev. 2018;31(4):e00020-18).

[0003] Staphylococcus aureus is a common foodborne pathogen that can cause a variety of infections and diseases in humans and animals, including skin infections, pneumonia, infectious arthritis, endocarditis, toxic shock syndrome, and sepsis. The pathogenic mechanism of Staphylococcus aureus is related to the various virulence factors produced by itself, including structural components of the pathogen (such as agglutinating factors, teichoic acid, etc.), enzyme substances, and toxins (such as leukocidin, hemolysin, shedding toxin, etc.). These virulence factors help bacteria adhere to host cells, obtain nutrients, and induce abnormal immune responses, thereby causing infection and poisoning reactions (Wu Yunpu et al., Experimental Animal Science, 2024;41(03):85-89). Chronic infections and antibiotic resistance induced by Staphylococcus aureus pose severe challenges to the public health field. In addition, Staphylococcus aureus is also an important pathogen that induces infectious diarrhea. It is of great value to use nutritional functional substances to fight against Staphylococcus aureus infections and the health problems caused by them.

[0004] Medium and long-chain triacylglycerols (MLCT) have attracted widespread attention as a star product among new structural lipids. The main structural feature of MLCT is that medium-chain fatty acids and long-chain fatty acids are simultaneously combined on a glycerol backbone. The fatty acid composition of MLCT is closer to breast milk, which is conducive to fat digestion and absorption, improves the absorption of lipid nutrients, inhibits the accumulation of body fat, and can quickly and stably supply energy (Cheng X et al., AnnuRev Food Sci Technol. 2024;15(1):381-408). Neuroacid (NA) is a core natural component of brain nerve fibers and nerve cells. It is a nutrient necessary for brain development and maintenance of function. It has biological functions such as repairing nerve ending activity and improving immunity. Currently, the biological functions of NA have received widespread attention from researchers.

[0005] There are no reports on the relationship between medium-chain triglycerides (MLCT) and neuraminic acid (NA) in fighting against Staphylococcus aureus infection and its induced diarrhea or other infections.

[0006] Therefore, there is still a need in the art for methods that can prevent or alleviate diarrhea, especially diarrhea caused by Staphylococcus aureus. Summary of the invention

[0007] In order to solve the above technical problems, the present invention studies the effect of the combination of MLCT and NA on anti-Staphylococcus aureus infection.

[0008] A first aspect of the present invention provides a nutritional composition comprising medium-chain fatty acid triglycerides and nervonic acid, wherein the mass ratio of the medium-chain fatty acid triglycerides to nervonic acid is 3:1 to 1500:1.

[0009] The applicant has found that when the nutritional composition contains medium- and long-chain fatty acid triglycerides and neuraminic acid in a defined mass ratio, there is a synergistic effect between the two in the following aspects: 1) preventing or alleviating diarrhea; and / or 2) inhibiting Staphylococcus aureus in vivo or in vitro; and / or 3) preventing or treating inflammation.

[0010] The second aspect of the present invention provides a food comprising the nutritional composition according to the first aspect of the present invention.

[0011] The third aspect of the present invention provides the use of the nutritional composition according to the first aspect of the present invention in preparing a product having one or more uses selected from 1) to 3):

[0012] 1) Prevent or relieve diarrhea;

[0013] 2) Inhibit Staphylococcus aureus in vivo or in vitro;

[0014] 3) Prevent or treat inflammation;

[0015] The nutritional composition comprises medium-chain and long-chain fatty acid triglycerides and nervonic acid.

[0016] Applicants have discovered that when a nutritional composition comprises medium- and long-chain fatty acid triglycerides and nervonic acid, the nutritional composition can: 1) prevent or alleviate diarrhea; and / or 2) inhibit Staphylococcus aureus in vivo or in vitro; and / or 3) prevent or treat inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Typical graphs showing fluorescence intensity in zebrafish intestine after single component sample treatment.

[0018] Figure 2 Statistical analysis results of zebrafish intestinal fluorescence intensity after single component and combination sample treatment, compared with the model control group. ** indicates P < 0.01, *** indicates P < 0.001.

[0019] Figure 3 Typical graphs showing fluorescence intensity in zebrafish intestine after single component and combination sample treatment.

[0020] Figure 4 Statistical analysis results of the fluorescence intensity of Staphylococcus aureus in zebrafish after treatment with single component and combination samples, compared with the model control group. ** indicates P < 0.01, *** indicates P < 0.001.

[0021] Figure 5 Typical graphs showing the fluorescence intensity of Staphylococcus aureus in zebrafish after single component and combination sample treatment.

[0022] Figure 6 The statistical analysis results of the number of neutrophils in the zebrafish intestine after treatment with single component and combination samples are shown, compared with the model control group. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0023] Figure 7 Typical graphs showing the number of neutrophils in the zebrafish intestine after treatment with single component and combination samples. DETAILED DESCRIPTION

[0024] The following definitions are provided to facilitate understanding of the present invention by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs. Preferred materials and methods are described herein, but any methods and materials similar to or equivalent to those described herein may be used in the practice of the present invention's tests. It should also be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to be limiting.

[0025] Definition of terms

[0026] Unless otherwise indicated or defined, all terms used have the usual meaning in the art, which will be understood by those skilled in the art. In addition, unless otherwise specified, all methods, steps, techniques and operations not specifically described in detail can and have been performed in a manner known per se, which will be understood by those skilled in the art.

[0027] As used herein, the term "medium-chain fatty acid" refers to a fatty acid having 6-12 carbon atoms in the carbon chain, such as caproic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, etc.; the term "long-chain fatty acid" refers to a fatty acid having more than 14 carbon atoms, generally 14-30 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" refer to the esterification reaction products of "medium-chain fatty acid" and "long-chain fatty acid" with glycerol, respectively; the term "medium-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. The present invention has no special requirements for the medium-long chain fatty acid triglyceride used, and the medium-long chain fatty acid triglyceride commonly used in the art can be used. The medium-chain fatty acid triglycerides can be used in pure form or in an impure form rich in medium-chain fatty acid triglycerides. For example, the medium-chain fatty acid triglycerides used in the present invention can contain C6-C 12 Fatty acid residues and C 14 -C 30 Fatty acid residue. C6-C 12 The fatty acid residue may be derived from one or more of the following: hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, etc. 14 -C 30The fatty acid residue may be 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, eicosapentaenoic acid, hexacosahexenoic acid, arachidonic acid, etc. In some embodiments, the C6-C 12 The fatty acid residue is derived from one or more of the following: caproic acid, caprylic acid, capric acid and lauric acid; and / or the C 14 -C 30 The fatty acid residue is derived from one or more of the following: myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid. 12 The fatty acid residues on the C 14 -C 30 The weight ratio of fatty acid residues is 0.124 to 2.000. In some embodiments, C6-C 12 The fatty acid residues on the C 14 -C 30 The weight ratio of fatty acid residues 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 any two thereof. In some embodiments, the "medium and long chain fatty acid triglyceride" can be prepared with reference to the method of patent CN115369132A.

[0028] As used herein, the term "nervonic acid" or "NA" is Nervonic acid in English, and its chemical name is cis-15-tetradecenoic acid, which is a ω-9 type monounsaturated fatty acid. Nervonic acid is present in high levels in nerve tissue and brain, and is an important substance that constitutes biological membranes and a marker of medulla in cerebrosides. It is also an "advanced nutrient" necessary for the growth, redevelopment and maintenance of nerve cells (especially brain cells, optic nerve cells, and peripheral nerves).

[0029] As used herein, "Staphylococcus aureus" is also called "golden Staphylococcus", which belongs to the genus Staphylococcus, is a representative of Gram-positive bacteria, is a common foodborne pathogenic microorganism, and is widely present in the natural environment. Under appropriate conditions, Staphylococcus aureus can produce enterotoxins and cause food poisoning. Enterotoxins are single-chain small molecule proteins with a molecular weight of about 26-29kDa, a relatively low molecular mass, and heat stability. They damage the human intestine and cause symptoms such as vomiting and diarrhea.

[0030] The present invention will be described in detail below.

[0031] Nutritional composition

[0032] A first aspect of the present invention provides a nutritional composition, comprising medium-chain fatty acid triglycerides (MLCT) and nervonic acid (NA), wherein the mass ratio of the medium-chain fatty acid triglycerides to nervonic acid is 3:1 to 1500:1.

[0033] The applicant has found that when a nutritional composition comprises medium- and long-chain fatty acid triglycerides and nervonic acid in a defined mass ratio, there is a synergistic effect between the two in the following aspects: preventing or alleviating diarrhea; and / or inhibiting Staphylococcus aureus in vivo or in vitro; and / or preventing or treating inflammation; for example, there is a synergistic effect in preventing or alleviating diarrhea, and optionally inhibiting Staphylococcus aureus in vivo or in vitro and / or preventing or treating inflammation.

[0034] In an embodiment, the mass ratio of the medium and long chain fatty acid triglycerides and neuraminic acid in the nutritional composition is 3:1 to 1500:1, for example, 3:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, 100:1, 150:1, 200:1, 300:1, 400:1, 500:1, 600:1, 700:1, 800:1, 900:1, 1000:1, 1100:1, 1200:1, 1300:1, 1400:1, 1500:1 or within the range defined by any two of them.

[0035] In some embodiments, the mass ratio of the medium-chain fatty acid triglycerides and neuraminic acid in the nutritional composition is 3:1 to 800:1; within the range, the synergistic effect of medium-chain fatty acid triglycerides and neuraminic acid in preventing or alleviating diarrhea is more significant, and at the same time, a synergistic effect is achieved in preventing or treating inflammation and, optionally, inhibiting Staphylococcus aureus in vivo or in vitro.

[0036] In some preferred embodiments, the mass ratio of the medium-chain fatty acid triglycerides and neuraminic acid in the nutritional composition is 3:1 to 150:1; within the range, the medium-chain fatty acid triglycerides and neuraminic acid achieve a synergistic effect in inhibiting Staphylococcus aureus in vivo or in vitro, and at the same time, the synergistic effect in preventing or treating inflammation is more significant.

[0037] In some preferred embodiments, the mass ratio of the medium-chain fatty acid triglycerides and neuraminic acid in the nutritional composition is 15:1 to 150:1; within the range, the synergistic effect of medium-chain fatty acid triglycerides and neuraminic acid in preventing or alleviating diarrhea is more significant; further preferably, the mass ratio of the two is 15:1 to 100:1, within the range, the synergistic effect of medium-chain fatty acid triglycerides and neuraminic acid in preventing or treating inflammation is more significant; further preferably, the mass ratio of the two is 15:1 to 70:1, within the range, the synergistic effect in preventing or treating inflammation is more significant; further preferably, the mass ratio of the two is 15:1 to 35:1, within the range, the synergistic effect of medium-chain fatty acid triglycerides and neuraminic acid in preventing or alleviating diarrhea is more significant.

[0038] In some preferred embodiments, the mass ratio of the medium-chain fatty acid triglycerides and neuraminic acid in the nutritional composition is 3:1 to 100:1, preferably 3:1 to 70:1, preferably 3:1 to 35:1, and preferably 3:1 to 15:1. Within each of the preferred ranges, the synergistic effect of medium-chain fatty acid triglycerides and neuraminic acid in inhibiting Staphylococcus aureus in vivo or in vitro is more significant.

[0039] In some embodiments, the nutritional composition consists of medium-chain fatty acid triglycerides and nervonic acid. In some embodiments, the diarrhea is bacterial diarrhea.

[0040] In some embodiments, the bacterial diarrhea is diarrhea caused by Staphylococcus aureus.

[0041] In some embodiments, the inflammation is bacterially caused inflammation.

[0042] In some embodiments, the inflammation is intestinal inflammation caused by Staphylococcus aureus.

[0043] food

[0044] A second aspect of the present invention relates to a food product comprising the nutritional composition of the first aspect of the present invention.

[0045] All the above descriptions about the first aspect of the present invention are applicable here and will not be repeated here.

[0046] In some embodiments, the food may be, for example, a functional food, a health food, or a health product.

[0047] In some embodiments, the food is a common food or a special food. As an example, the food can be selected from dairy products, candies, beverages, bread and biscuits, for example, the food can be selected from milk powder or fermented food. In some embodiments, the milk powder is infant formula milk powder.

[0048] In some embodiments, the food is an infant food such as an infant formula.

[0049] In some embodiments, the food product may be in the form of a powder, a tablet, or a liquid.

[0050] In some embodiments, based on the dry basis weight of the food (after deducting water), the content of nervonic acid may be 0.002% to 0.3%. As an example, based on the weight of the product, the content of nervonic acid may be 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.25, 0.30 dry basis weight%, or within the range defined by any two of them. The dry basis weight of medium and long chain fatty acid triglycerides (after deducting water) can be calculated according to the ratio of the two.

[0051] In some embodiments, based on the dry weight of the food (after deducting water), the content of medium and long chain fatty acid triglycerides can be 0.0004% to 30.0%. As an example, based on the weight of the food, the content of medium and long chain fatty acid triglycerides can be 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0. 0, 29.0, 30.0, 31.0, 32.0, 33.0, 34.0, 35.0, 36.0, 37.0, 38.0, 39.0, 40.0, 41.0, 42.0, 43.0, 44.0, 45.0, 46.0, 47.0, 48.0, 49.0, 50.0, 51.0, 52.0, 53.0, 54.0, 55.0, 56.0, 57.0, 58.0, 59.0, 60.0, 61.0, 62.0, 63.0, 64.0, 65.0, 66.0, 67.0, 68.0, 69.0, 70.0, 71.0, 72.0, 73.0

[0052] 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 also includes one or more selected from the following: raw milk, desalted whey powder, concentrated whey protein, lactose, edible vegetable blending oil, oligofructose, oligogalactose, nucleotides, choline, vitamins, minerals, DHA and taurine.

[0053] For example, when the food is milk powder, in addition to the nutritional composition described in the first aspect of the present invention, the milk powder may also include proteins such as α-lactalbumin and milk fat globule membrane protein; carbohydrates such as lactose; lipids; minerals such as calcium, iron, phosphorus, etc.; vitamins; other additives such as whey powder, choline bitartrate, docosahexaenoic acid, arachidonic acid, walnut oil, etc.

[0054] The food can be produced by a preparation method commonly used in the art, which will not be described in detail here.

[0055] application

[0056] The third aspect of the present invention relates to the use of the nutritional composition described in the first aspect of the present invention in the preparation of a product having one or more uses selected from 1) to 3) (e.g., non-therapeutic uses):

[0057] 1) Prevent or relieve diarrhea;

[0058] 2) Inhibit Staphylococcus aureus in vivo or in vitro; 3) Prevent or treat inflammation;

[0059] The nutritional composition comprises medium-chain and long-chain fatty acid triglycerides and nervonic acid.

[0060] The applicant has found that when a nutritional composition contains medium- and long-chain fatty acid triglycerides and nervonic acid, the nutritional composition can achieve one or more of the above uses 1)-3) (for example 1 alone), 2 alone), 3 alone), 1)+2), 1)+3), 2)+3), or 1)+2)+3)); preferably the above uses 1)-3).

[0061] All the above descriptions about the first aspect of the present invention are applicable here and will not be repeated here.

[0062] In some embodiments, the nutritional composition or a product made therefrom is for oral administration to humans.

[0063] In some embodiments, when the composition is orally taken by humans, the dosage of nervonic acid is 2-300 mg / day, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300 mg / day, or a range defined by any two of them. Preferably, the dosage of medium- and long-chain fatty acid triglycerides can be calculated based on the ratio of medium- and long-chain fatty acid triglycerides to nervonic acid described for the nutritional composition of the first aspect of the present invention.

[0064] In some embodiments, when the composition is orally administered to humans, the dosage of medium- and long-chain fatty acid triglycerides is 0.0004-30.0 g / day, for example, 0.0004 g / day, 0.001 g / day, 0.005 g / day, 0.01 g / day, 0.02 g / day, 0.05 g / day, 0.1 g / day, 0.2 g / day, 0.3 g / day, 0.4 g / day, 0.5 g / day, 0.6 g / day, 0.7 g / day, 0.8 g / day, 0.9 g / day, 1 g / day, 2 g / day, 3 g / day, 4 g / day, 5 g / day, 6 g / day, 7 g / day, 8 g / day, 9 g / day, 10 g / day, 11 g / day, 12 g / day, 13 g / day, 14 g / day, 15 g / day, 16 g / day, 17 g / day, g / day, 18 g / day, 19 g / day, 20 g / day, 21 g / day, 22 g / day, 23 g / day, 24 g / day, 25 g / day, 26 g / day, 27 g / day, 28 g / day, 29 g / day, or 30 g / day or any range therebetween.

[0065] In some embodiments, the product is a food, a medicine or a kit. For the food, the above description of the second aspect of the present invention is applicable here and will not be repeated here.

[0066] In some embodiments, the drug can be in any dosage form such as a solid preparation (powder, tablet, etc.) or a liquid preparation.

[0067] In some embodiments, depending on the dosage form, the drug further comprises a pharmaceutically acceptable carrier and / or excipient.

[0068] Example

[0069] The present invention will be more easily understood with reference to the following examples, which are merely used to illustrate some aspects and embodiments of the present invention and are not intended to limit the present invention.

[0070] Unless otherwise stated, the reagents used in this embodiment are all commercially available materials or conventional materials. The medium-chain fatty acid triglycerides in all the following embodiments are derived from medium-chain fatty acid edible oil (purity of 69%), produced by Qingdao Haizhiyuan Life Science Co., Ltd., with a production batch number of Y1505-22120101; the purity of nervonic acid is more than 90%, purchased from Guangzhou Green Cui Biology, with a production batch number of FH20240120.

[0071] Example 1 Evaluation of the efficacy of a single sample in improving diarrhea

[0072] Wild-type AB strain zebrafish at 4 days post-fertilization (4 dpf) were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations are shown in Table 1), and the positive control vancomycin concentration was 1000 μg / mL. At the same time, a normal control group (no treatment) and a model control group (diarrhea model, no medication) were set up, and the volume of each well was 3 mL. After treatment at 28℃ for 8 h, each experimental group was given water-soluble Nile red as a fluorescent indicator of intestinal contents, and fed overnight. The next day, the samples and Nile red were washed off, and the samples were continued to be given water-soluble (concentrations are shown in Table 1). Except for the normal control group, the rest of the experimental groups were given water-soluble Staphylococcus aureus to establish a zebrafish diarrhea model. After treatment at 28℃ for 30 h, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. NIS-Elements D 3.20 advanced image processing software was used to analyze and collect data, and the intensity of zebrafish intestinal fluorescence signals was analyzed. The statistical analysis results of this index were used to evaluate the efficacy of the samples in improving diarrhea induced by Staphylococcus aureus. The statistical analysis results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and P<0.05 indicated that the difference was statistically significant.

[0073] According to the above method, medium-chain fatty acid triglycerides and neuraminic acid were administered to zebrafish at different doses. The two single components improved diarrhea. The higher the intestinal fluorescence intensity, the lower the degree of diarrhea. The dosage and results are shown in Table 1.

[0074] Table 1 Results of the efficacy evaluation experiment on the efficacy of single component in improving diarrhea (intestinal fluorescence intensity)

[0075]

[0076] Note: P value indicates the significant difference in intestinal fluorescence intensity between the medium-chain fatty acid triglyceride group or neuraminic acid group and the model control group.

[0077] From Table 1, Figure 1The results showed that the intestinal fluorescence intensity of 4 dpf wild-type AB strain zebrafish was significantly decreased after induction with Staphylococcus aureus (model control group) compared with the normal control group (P < 0.001), indicating that the diarrhea model induced by Staphylococcus aureus was successfully established. The positive control (vancomycin), monomeric medium-chain fatty acid triglycerides (125 µg / mL, 250 µg / mL, 500 µg / mL, 750 µg / mL) and neuraminic acid (15.6 µg / mL, 62.5 µg / mL, 250 µg / mL) all had a significant effect on improving diarrhea, as shown by the stronger intestinal fluorescence intensity of zebrafish compared with the model control group (P < 0.05). The effect of monomeric medium-chain fatty acid triglycerides on improving diarrhea was the strongest at a concentration of 250 µg / mL. When the concentration was further increased (500 µg / mL, 750 µg / mL), the effect of improving diarrhea became weaker or similar. Therefore, 250 µg / mL was the optimal concentration of medium-chain fatty acid triglycerides for improving diarrhea. The effect of monomeric nervonic acid on improving diarrhea was the strongest at a concentration of 62.5 µg / mL. When the concentration was further increased (250 µg / mL), the effect of zebrafish on improving diarrhea became weaker. Therefore, 62.5 µg / mL was the optimal concentration of nervonic acid for improving diarrhea.

[0078] Example 2 Evaluation of the efficacy of the composition in improving diarrhea

[0079] 4 dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). Samples of MLCT and neuraminic acid combination were given water-soluble (concentrations shown in Table 2), and the positive control vancomycin concentration was 1000 μg / mL. At the same time, a normal control group and a model control group were set up, and the volume of each well was 3 mL. After 8 h of treatment at 28℃, each experimental group was given Nile red as a fluorescent indicator of intestinal contents in water-soluble form, and fed overnight. The next day, the samples and Nile red were washed off, and the samples were continued to be given water-soluble form (concentrations shown in Table 2). Except for the normal control group, the rest of the experimental groups were given Staphylococcus aureus in water-soluble form to establish a zebrafish diarrhea model. After 30 h of treatment at 28℃, 10 zebrafish were randomly selected from each experimental group and placed under a fluorescence microscope for photography. The data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software, and the intensity of zebrafish intestinal fluorescence signals was analyzed. The results of statistical analysis of this index were used to evaluate the efficacy of the samples in improving diarrhea induced by Staphylococcus aureus. The statistical results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and P < 0.05 indicated that the difference was statistically significant.

[0080] According to the above method, medium-chain fatty acid triglycerides and neuraminic acid were administered to zebrafish at different dosage ratios, and the efficacy of the two single components in improving diarrhea (intestinal fluorescence intensity) was shown in Table 2. The administration results are shown in Table 2. Figure 2 , Figure 3 shown.

[0081] Table 2 Results of efficacy evaluation experiment on the efficacy of the composition in improving diarrhea (intestinal fluorescence intensity)

[0082]

[0083] Note: P value indicates the significant difference in intestinal fluorescence intensity between the medium-chain fatty acid triglyceride group or neuraminic acid group and the model control group.

[0084] From Table 2, Figure 2 , Figure 3 The results showed that compared with the model control group, the compositions of Examples 1 to 11 all had the effect of improving diarrhea, which was specifically manifested by the enhancement of intestinal fluorescence intensity (P<0.05). Compared with the effect at the optimal concentration of MLCT and neuraminic acid (μg / mL), the values ​​of zebrafish intestinal fluorescence intensity in Examples 2 to 10 were significantly enhanced, indicating that MLCT and neuraminic acid had a synergistic effect at a ratio of (3-1500):1; among them, the values ​​of zebrafish intestinal fluorescence intensity in Examples 2 to 9 where the ratio of MLCT to neuraminic acid was (3-800):1 were higher, indicating that the synergistic effect of MLCT and neuraminic acid at a ratio of (3-800):1 was more significant; the values ​​of zebrafish intestinal fluorescence intensity in Examples 2 to 7 where the ratio of MLCT to neuraminic acid was (3-150:1) were further improved, indicating that the synergistic effect of MLCT and neuraminic acid at a ratio of (3-150:1) was further enhanced; the values ​​of zebrafish intestinal fluorescence intensity in Examples 3 to 7 where the ratio of MLCT to neuraminic acid was (15-150:1) were even further enhanced, indicating that the synergistic effect of MLCT and neuraminic acid at a ratio of (15-150:1) was further enhanced.

[0085] Example 3 Experiment on the efficacy of inhibiting Staphylococcus aureus

[0086] 4 dpf wild-type AB strain zebrafish were randomly selected in 6-well plates, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations are shown in Table 3), and the positive control vancomycin concentration was 1000 μg / mL. At the same time, a normal control group and a model control group were set up, and the volume of each well was 3 mL. After 24 h of treatment at 28℃. Except for the normal control group, the rest of the experimental groups were given water-soluble Staphylococcus aureus to establish the zebrafish Staphylococcus aureus infection model. After CM-DiI was used to label Staphylococcus aureus for 6 h at 28℃, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-ElementsD 3.20 advanced image processing software, and the fluorescence intensity of Staphylococcus aureus in the zebrafish intestine was analyzed. The antibacterial efficacy of the samples was evaluated by statistical analysis of the index. The statistical analysis results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0087] The dosages and results are shown in Table 3.

[0088] Table 3 Results of the anti-Staphylococcus aureus efficacy evaluation experiment

[0089]

[0090] Note: P value indicates the significant difference in fluorescence intensity of Staphylococcus aureus between the medium-chain fatty acid triglyceride group or neuraminic acid group and the model control group.

[0091] From Table 3, Figure 4 , Figure 5 The results showed that compared with the model control group induced by Staphylococcus aureus, the positive control (vancomycin), monomeric medium-chain fatty acid triglycerides (250 µg / mL), neuraminic acid (62.5 µg / mL) and the combination of formulas 1-11 all had significant anti-Staphylococcus aureus efficacy, which was specifically manifested in that the fluorescence intensity of Staphylococcus aureus in zebrafish was weakened compared with the model control group (P<0.05). Compared with monomeric medium-chain fatty acid triglycerides (250 µg / mL) and neuraminic acid (62.5 µg / mL), formula 2-7 with a ratio of MLCT to NA of (3-150):1 had stronger anti-Staphylococcus aureus efficacy, as reflected in the more obvious weakening of the fluorescence intensity of S. aureus, indicating a synergistic effect; among them, formula 2-6 with a ratio of MLCT to NA of (3-100):1 had stronger anti-Staphylococcus aureus efficacy, indicating a more significant synergistic effect; formula 2-5 with a ratio of (3-66):1 had even further enhanced anti-Staphylococcus aureus efficacy, indicating a further enhanced synergistic effect.

[0092] Example 4 Evaluation of the anti-inflammatory efficacy of the composition

[0093] 4 dpf transgenic neutrophil green fluorescent zebrafish (MPX) were randomly selected in 6-well plates, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble (concentrations shown in Table 4), and the positive control vancomycin concentration was 1000 μg / mL. At the same time, a normal control group and a model control group were set up, and the volume of each well was 3 mL. After treatment at 28℃ for 24 h, Staphylococcus aureus was given water-soluble in all experimental groups except the normal control group to establish a zebrafish inflammation model. After treatment at 28℃ for 6 h, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software, and the number of zebrafish intestinal neutrophils was analyzed. The statistical analysis results of this indicator were used to evaluate the efficacy of the sample in relieving inflammation induced by Staphylococcus aureus. The statistical analysis results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and P < 0.05 indicated that the difference was statistically significant.

[0094] According to the above method, medium-chain fatty acid triglycerides and neuraminic acid were administered to zebrafish at different dosage ratios to detect their anti-inflammatory effects (number of intestinal neutrophils). The dosages and administration results are shown in Table 4.

[0095] Table 4 Results of the anti-inflammatory efficacy evaluation test of the composition

[0096]

[0097] Note: P value is the significant difference between each embodiment group and the model control group

[0098] Figure 7 A typical graph showing the number of neutrophils in the zebrafish intestine after sample treatment.

[0099] Table 4 Figure 6 and Figure 7 The results showed that compared with the model control group, the combination of MLCT and neuraminic acid in the ratio of (0.2-800):1, the combination of 1-9, had anti-inflammatory effects, as shown by a decrease in the number of intestinal neutrophils ( P<0.05). Compared with the optimal concentrations of MLCT (concentration 250 µg / mL) and neuraminic acid (concentration 62.5 µg / mL), the number of zebrafish intestinal neutrophils in combinations 2-9 decreased, indicating that MLCT and neuraminic acid had a synergistic effect at a ratio of (3-800):1; among them, the number of zebrafish intestinal neutrophils in combinations 2-7 was even lower, indicating that the synergistic effect of MLCT and neuraminic acid at a ratio of (3-150):1 was more significant; the number of zebrafish intestinal neutrophils in combinations 3-6 was further reduced, indicating that the synergistic effect of MLCT and neuraminic acid at a ratio of (15-100):1 was further enhanced; the number of zebrafish intestinal neutrophils in combinations 3-5 was even further reduced, indicating that the synergistic effect of MLCT and neuraminic acid at a ratio of (15-66):1 was even further enhanced.

[0100] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A nutritional composition comprising medium-chain fatty acid triglycerides and nervonic acid, wherein the mass ratio of the medium-chain fatty acid triglycerides to nervonic acid is 3:1 to 1500:

1. 2 . The nutritional composition according to claim 1 , wherein the mass ratio of the medium- and long-chain fatty acid triglycerides to nervonic acid is 3:1 to 800:

1. 3 . The nutritional composition according to claim 1 , wherein the mass ratio of the medium- and long-chain fatty acid triglycerides to nervonic acid is 3:1 to 150:

1.

4. Food comprising the nutritional composition according to any one of claims 1 to 3.

5. The food according to claim 4, which is selected from one or more of dairy products, candies, beverages, bread and biscuits. The food according to claim 4 , which is milk powder or fermented food. The food according to claim 4 , which is infant food. The food according to claim 6 , wherein the milk powder is infant formula milk powder.

9. Use of the nutritional composition according to any one of claims 1 to 3 in preparing a product having one or more uses selected from 1) to 3): 1) Prevent or relieve diarrhea; 2) Inhibit Staphylococcus aureus in vivo or in vitro; 3) Prevent or treat inflammation; The nutritional composition comprises medium-chain and long-chain fatty acid triglycerides and nervonic acid.

10. The use according to claim 9, wherein the diarrhea is bacterial diarrhea and / or the inflammation is inflammation caused by bacteria.

11. The use according to claim 10, wherein the bacterial diarrhea is diarrhea caused by Staphylococcus aureus and / or the inflammation is intestinal inflammation caused by Staphylococcus aureus.

12. The use according to claim 9, wherein the product is a medicine or a kit.

13. The use according to claim 12, wherein the dosage form of the drug is selected from one or more of tablets, granular powders, capsules and liquids.

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