Nutritional composition, product containing the same, and use of the nutritional composition

Abstract

The present invention relates to a nutritional composition. The nutritional composition comprises medium- and long-chain triglycerides and nervonic acid in a mass ratio of 3:1 to 1500:1, which have a synergistic effect in preventing or alleviating diarrhea, inhibiting Staphylococcus aureus in vivo or in vitro, and / or preventing or treating inflammation. The present invention also relates to a product comprising the nutritional composition. The present invention also relates to the use of the nutritional composition in the preparation of a product having one or more uses selected from 1) to 3): 1) preventing or alleviating diarrhea; 2) inhibiting Staphylococcus aureus in vivo or in vitro; 3) preventing or treating inflammation.

Description

Technical Field

[0001] The present disclosure relates to nutritional compositions, products containing the same, and applications of the nutritional compositions. Background Art

[0002] Staphylococcus aureus is a common foodborne pathogenic microorganism that can cause various 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 a variety of virulence factors produced by itself, including structural components of the pathogen (such as clumping factor, teichoic acid, etc.), enzyme substances, and toxins (such as leukocidin, hemolysin, exfoliative toxin, etc.). These virulence factors contribute to bacterial adhesion to host cells, nutrient acquisition, and the induction of abnormal immune responses, thereby causing infection and toxic reactions (Wu Yunpu et al., Laboratory Animal Science, 2024; 41(03): 85 - 89). The chronic infections and antibiotic resistance induced by Staphylococcus aureus pose a severe challenge to the public health field. In addition, Staphylococcus aureus is also an important pathogen causing infectious diarrhea. It has important value to combat Staphylococcus aureus infection and the resulting health problems through nutritional functional substances.

[0003] Medium and long-chain triacylglycerols (MLCT), as a star product among novel structured lipids, have received extensive attention. The main structural feature of MLCT is that a glycerol backbone is simultaneously combined with medium-chain fatty acids and long-chain fatty acids. The fatty acid composition of MLCT is closer to breast milk, which is beneficial to fat digestion and absorption, improves the absorption of lipid nutrients, inhibits the accumulation of body fat, and can provide energy quickly and stably (Cheng X et al., Annual Review of Food Science and Technology, 2024; 15(1): 381 - 408). Nervonic acid (NA) is a core natural component of brain nerve fibers and nerve cells and is a nutrient necessary for brain development and maintaining function. It has biological functions such as repairing the activity of nerve endings and enhancing immunity. Currently, the biological functions of NA have received extensive attention from researchers.

[0004] The relationship between medium and long-chain triacylglycerols (MLCT), nervonic acid (NA) and combating Staphylococcus aureus infection and the diarrhea or other infections induced by it has not been reported.

[0005] Therefore, there is still a need in the art for methods that can prevent or relieve diarrhea, especially diarrhea caused by Staphylococcus aureus. Summary of the Invention

[0006] To solve the above technical problems, the present invention studied the effect of the composition of MLCT and NA against Staphylococcus aureus infection.

[0007] In a first aspect, the present invention provides a nutritional composition comprising medium and long chain triglycerides and nervonic acid, wherein the mass ratio of the medium and long chain triglycerides to nervonic acid is from 3:1 to 1500:1.

[0008] The applicant has found that when the nutritional composition comprises medium and long chain triglycerides and nervonic acid in the 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.

[0009] In a second aspect, the present invention provides a product comprising the nutritional composition according to the first aspect of the present invention.

[0010] In a third aspect, the present invention provides the use of the nutritional composition according to the first aspect of the present invention in the preparation of a product having one or more of the uses selected from 1) to 3):

[0011] 1) Preventing or alleviating diarrhea;

[0012] 2) Inhibiting Staphylococcus aureus in vivo or in vitro;

[0013] 3) Preventing or treating inflammation;

[0014] wherein the nutritional composition comprises medium and long chain triglycerides and nervonic acid.

[0015] The applicant has found that when the nutritional composition comprises medium and long chain 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. Description of the Drawings

[0016] Figure 1 Typical diagram showing the fluorescence intensity of the zebrafish intestine after treatment with single-component samples.

[0017] Figure 2 Showing the statistical analysis results of the fluorescence intensity of the zebrafish intestine after treatment with single-component and composition samples, compared with the model control group. ** indicates P<0.01, *** indicates P<0.001.

[0018] Figure 3 Typical diagram showing the fluorescence intensity of the zebrafish intestine after treatment with single-component and composition samples.

[0019] Figure 4Show the statistical analysis results of the fluorescence intensity of Staphylococcus aureus in zebrafish after treatment with single-component and composition samples, compared with the model control group. ** indicates P < 0.01, and *** indicates P < 0.001.

[0020] Figure 5 Show the typical graphs of the fluorescence intensity of Staphylococcus aureus in zebrafish after treatment with single-component and composition samples.

[0021] Figure 6 Show the statistical analysis results of the number of neutrophils in the zebrafish intestine after treatment with single-component and composition samples, compared with the model control group. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0022] Figure 7 Show the typical graphs of the number of neutrophils in the zebrafish intestine after treatment with single-component and composition samples. Detailed implementation manners

[0023] 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 meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Preferred materials and methods are described herein, but any methods and materials similar to or equivalent to those described herein can be used in the practice of testing the present invention. It should also be understood that the terms used herein are for the purpose of describing specific embodiments only and are not intended to be limiting.

[0024] Term definitions

[0025] Unless otherwise indicated or defined, all terms used have their ordinary meanings in the art, which will be understood by those skilled in the art. In addition, unless otherwise stated, all methods, steps, techniques, and operations not specifically detailed can and have been carried out in a manner known per se, which will be understood by those skilled in the art.

[0026] As used herein, 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, lauric 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. 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. For example, the medium and long-chain fatty acid triglycerides used in the present invention may contain C6-C 12 fatty acid residues and C 14 -C 30 fatty acid residues. The C6-C 12 fatty acid residues may be derived from one or more of the following: caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, etc. C 14 -C 30 fatty acid residues may be derived from one or more of the following: myristic acid, palmitic acid, margaric 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. In some embodiments, the C6-C 12 fatty acid residues are derived from one or more of the following: caproic acid, caprylic acid, capric acid, and lauric acid; and / or the C 14 -C 30 fatty acid residues are 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. In some embodiments, the weight ratio of the C6-C 12 fatty acid residues to the C 14 -C 30 fatty acid residues is from 0.124 to 2.000. In some embodiments, the weight ratio of the C6-C 12 fatty acid residues to the C 14 -C 30The 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. In some embodiments, the "medium-chain and long-chain fatty acid triglyceride" can be prepared by referring to the method of Patent CN115369132A.

[0027] As used herein, the term "nervonic acid" or "NA", the English name is Nervonic acid, and the chemical name is cis-15-tetracosenoic acid, which is a monounsaturated fatty acid of the ω-9 type. Nervonic acid is relatively abundant in nerve tissues and the brain, is an important component of biological membranes, and is a marker of medulla in cerebrosides. It is also an essential "advanced nutrient" for the growth, redevelopment, and maintenance of nerve cells (especially brain cells, optic nerve cells, and peripheral nerves).

[0028] As used herein, "Staphylococcus aureus" is also called "S. aureus", belongs to the genus Staphylococcus, is a representative of Gram-positive bacteria, and is a common foodborne pathogenic microorganism that widely exists in the natural environment. Under appropriate conditions, Staphylococcus aureus can produce enterotoxin, causing food poisoning. Enterotoxin is a single-chain small molecule protein with a molecular weight of about 26-29 kDa, a relatively low relative molecular mass, heat stability, and can damage the human intestine, resulting in symptoms such as vomiting and diarrhea.

[0029] The present invention will be specifically described below.

[0030] Nutritional composition

[0031] In a first aspect of the present invention, there is provided a nutritional composition comprising medium-chain and long-chain fatty acid triglyceride (MLCT) and nervonic acid (NA), wherein the mass ratio of the medium-chain and long-chain fatty acid triglyceride to the nervonic acid is from 3:1 to 1500:1.

[0032] The applicant has found that when the nutritional composition contains medium-chain and long-chain fatty acid triglyceride and nervonic acid in the 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 in inhibiting Staphylococcus aureus in vivo or in vitro and / or preventing or treating inflammation.

[0033] In an embodiment, the mass ratio of the medium and long-chain fatty acid triglyceride to nervonic acid in the nutritional composition is from 3:1 to 1500:1, such as 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.

[0034] In some embodiments, the mass ratio of the medium and long-chain fatty acid triglyceride to nervonic acid in the nutritional composition is from 3:1 to 800:1; within this range, the synergistic effect of the medium and long-chain fatty acid triglyceride and nervonic 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 also in inhibiting Staphylococcus aureus in vivo or in vitro.

[0035] In some preferred embodiments, the mass ratio of the medium and long-chain fatty acid triglyceride to nervonic acid in the nutritional composition is from 3:1 to 150:1; within this range, a synergistic effect is achieved in inhibiting Staphylococcus aureus in vivo or in vitro by the medium and long-chain fatty acid triglyceride and nervonic acid, and at the same time, the synergistic effect in preventing or treating inflammation is more significant.

[0036] In some preferred embodiments, the mass ratio of the medium and long-chain fatty acid triglyceride to nervonic acid in the nutritional composition is from 15:1 to 150:1; within this range, the synergistic effect of the medium and long-chain fatty acid triglyceride and nervonic acid in preventing or alleviating diarrhea is more significant; further preferably, the mass ratio of the two is from 15:1 to 100:1, within which the synergistic effect of the medium and long-chain fatty acid triglyceride and nervonic acid in preventing or treating inflammation is more significant; further preferably, the mass ratio of the two is from 15:1 to 70:1, within which the synergistic effect in preventing or treating inflammation is more significant; further preferably, the mass ratio of the two is from 15:1 to 35:1, within which the synergistic effect of the medium and long-chain fatty acid triglyceride and nervonic acid in preventing or alleviating diarrhea is more significant.

[0037] In some preferred embodiments, the mass ratio of the medium- and long-chain fatty acid triglyceride to nervonic acid in the nutritional composition is from 3:1 to 100:1, preferably from 3:1 to 70:1, preferably from 3:1 to 35:1, preferably from 3:1 to 15:1. Within each of the preferred ranges, the synergistic effect of the medium- and long-chain fatty acid triglyceride and nervonic acid in inhibiting Staphylococcus aureus in vivo or in vitro is more significant.

[0038] In some embodiments, the nutritional composition consists of a medium- and long-chain fatty acid triglyceride and nervonic acid. In some embodiments, the diarrhea is bacterial diarrhea.

[0039] In some embodiments, the bacterial diarrhea is diarrhea caused by Staphylococcus aureus.

[0040] In some embodiments, the inflammation is inflammation caused by bacteria.

[0041] In some embodiments, the inflammation is intestinal inflammation caused by Staphylococcus aureus.

[0042] Product

[0043] The second aspect of the present invention relates to a product comprising the nutritional composition of the first aspect of the present invention.

[0044] All of the above descriptions regarding the first aspect of the present invention apply herein and will not be repeated here.

[0045] In some embodiments, the product can be in the form of a powder, tablet or liquid.

[0046] In some embodiments, based on the dry weight (after deducting moisture) of the product, the content of nervonic acid can be from 0.002% to 0.3%. By way of example, based on the weight of the product, the content of nervonic acid can 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 weight %, or within the range defined by any two of them. The dry weight (after deducting moisture) of the medium- and long-chain fatty acid triglyceride can be calculated according to the ratio of the two.

[0047] In some embodiments, based on the dry basis weight of the product (after deducting moisture), the content of medium and long chain fatty acid triglycerides can be from 0.0004% to 30.0%. As an example, based on the weight of the product, 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, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, 12.0, 13.0, 14.0, 15.0, 16.0, 17.0, 18.0, 19.0, 20.0, 21.0, 22.0, 23.0, 24.0, 25.0, 26.0, 27.0, 28.0, 29.0, 30.0 dry basis weight %, or within the range defined by any two of them.

[0048] 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 product further includes one or more selected from the following: raw cow milk, demineralized whey powder, concentrated whey protein, lactose, edible vegetable blended oil, fructooligosaccharide, galactooligosaccharide, nucleotide, choline, vitamin, mineral, DHA and taurine.

[0049] The product can be produced by the preparation methods commonly used in the art, which will not be elaborated here.

[0050] Application

[0051] The third aspect of the present invention relates to the application 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) (for example, non-therapeutic applications):

[0052] 1) Preventing or alleviating diarrhea;

[0053] 2) Inhibiting Staphylococcus aureus in vivo or in vitro; 3) Preventing or treating inflammation;

[0054] wherein the nutritional composition contains medium and long chain fatty acid triglycerides and nervonic acid.

[0055] The applicant has found that when the 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).

[0056] All of the above descriptions regarding the first aspect of the present invention apply here and will not be repeated herein.

[0057] In some embodiments, the nutritional composition or the product made therefrom is for oral use in humans.

[0058] In some embodiments, when the composition is orally administered to a human, the dosage of nervonic acid is 2-300 mg / day, such as 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 the range defined by any two of them. Preferably, the dosage of medium and long chain fatty acid triglycerides can be calculated according to 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.

[0059] In some embodiments, when the composition is orally administered to a human, the dosage of medium and long chain fatty acid triglycerides is 0.0004-30.0 g / day, such as 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, 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.

[0060] In some embodiments, the product is a drug. For the product, all of the above descriptions regarding the second aspect of the present invention apply here and will not be repeated herein.

[0061] In some embodiments, the drug can be in any dosage form such as solid preparations (powders, tablets, etc.) or liquid preparations.

[0062] In some embodiments, according to the dosage form, the drug further comprises a pharmaceutically acceptable carrier and / or excipient.

[0063] Examples

[0064] The present invention will be more easily understood with reference to the following examples, which are only used to illustrate some aspects and embodiments of the present invention and are not intended to limit the present invention.

[0065] Unless otherwise stated, the reagents used in this example are all commercially available materials or conventional materials. The medium and long-chain fatty acid triglycerides in all the following examples are derived from medium and long-chain fatty acid edible oil (purity 69%), produced by Qingdao Haizhiyuan Life Technology Co., Ltd., production batch number Y1505 - 22120101; the purity of nervonic acid is more than 90%, purchased from Guangzhou Green Extract Biotechnology, production batch number FH20240120.

[0066] Example 1 Evaluation of the Efficacy of a Single Sample in Improving Diarrhea

[0067] Wild-type AB strain zebrafish at 4 days post-fertilization (4 dpf) were randomly selected and placed in a 6-well plate, with 30 zebrafish in each well (experimental group). The samples were administered by dissolving in water (concentrations are shown in Table 1), the positive control vancomycin at a concentration of 1000 μg / mL, and a normal control group (without any treatment) and a model control group (diarrhea model, without drug administration) were set up simultaneously, with a volume of 3 mL in each well. After treatment at 28°C for 8 h, Nile red was administered by dissolving in water as a fluorescent indicator for intestinal contents in each experimental group, and the zebrafish were fed overnight. The next day, the samples and Nile red were washed off, and the samples were continued to be administered by dissolving in water (concentrations are shown in Table 1). Except for the normal control group, a diarrhea model of zebrafish was established by administering Staphylococcus aureus dissolved in water in the remaining experimental groups. After treatment at 28°C for 30 h, 10 zebrafish were randomly selected from each experimental group and placed under a fluorescence microscope for photographing, and data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The fluorescence signal intensity of the zebrafish intestine was analyzed, and the efficacy of the samples in improving diarrhea induced by Staphylococcus aureus was evaluated based on the statistical analysis results of this index. The statistical processing results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software, and P < 0.05 indicates that the difference is statistically significant.

[0068] According to the above method, medium and long-chain fatty acid triglycerides and nervonic acid were administered to zebrafish at different doses respectively. The higher the intestinal fluorescence intensity, the lower the degree of diarrhea, indicating the efficacy of the two single components in improving diarrhea. The administration doses and results are shown in Table 1.

[0069] Table 1 Experimental Results of the Efficacy Evaluation of a Single Component in Improving Diarrhea Efficacy (Intestinal Fluorescence Intensity)

[0070]

[0071] Note: The P value is the significance of the difference in intestinal fluorescence intensity between the medium and long-chain fatty acid triglyceride group or nervonic acid group, etc. and the model control group.

[0072] From Table 1, Figure 1 the results show that after 4dpf wild-type AB strain zebrafish were induced by Staphylococcus aureus (model control group), compared with the normal control group, the intestinal fluorescence intensity decreased significantly (P < 0.001), indicating that the diarrhea model induced by Staphylococcus aureus was successfully established. The positive control (vancomycin), monomeric medium and long-chain fatty acid triglycerides (125 μg / mL, 250 μg / mL, 500 μg / mL, 750 μg / mL) and nervonic acid (15.6 μg / mL, 62.5 μg / mL, 250 μg / mL) all had a significant effect on improving diarrhea. Specifically, compared with the model control group, the intestinal fluorescence intensity of zebrafish became stronger (P < 0.05). The monomeric medium and long-chain fatty acid triglyceride had the strongest effect on improving diarrhea at a concentration of 250 μg / mL. When its concentration was further increased (500 μg / mL, 750 μg / mL), its effect on improving diarrhea became weaker or similar. Therefore, the concentration of 250 μg / mL was the optimal concentration for the monomeric medium and long-chain fatty acid triglyceride to improve diarrhea efficacy. The monomeric nervonic acid had the strongest effect on improving diarrhea at a concentration of 62.5 μg / mL. When its concentration was further increased (250 μg / mL), the effect of zebrafish on improving diarrhea became weaker instead. Therefore, 62.5 μg / mL was the optimal concentration for nervonic acid to improve diarrhea efficacy.

[0073] Example 2 Efficacy Evaluation of the Composition in Improving Diarrhea

[0074] Wild-type AB strain zebrafish at 4 dpf were randomly selected and placed in a 6-well plate, with 30 zebrafish in each well (experimental group). Samples of the MLCT and nervonic acid composition were administered in water (concentrations are shown in Table 2), the positive control was vancomycin at a concentration of 1000 μg / mL. At the same time, a normal control group and a model control group were set up, and the volume per well was 3 mL. After treatment at 28 °C for 8 h, Nile red was administered in water as a fluorescent indicator for intestinal contents to each experimental group, and the fish were fed overnight. The next day, the samples and Nile red were washed off, and the samples were continued to be administered in water (concentrations are shown in Table 2). Except for the normal control group, Staphylococcus aureus was administered in water to the remaining experimental groups to establish a zebrafish diarrhea model. After treatment at 28 °C for 30 h, 10 zebrafish were randomly selected from each experimental group and placed under a fluorescence microscope for photographing. The NIS-Elements D 3.20 advanced image processing software was used to analyze and collect data, and the fluorescence signal intensity of the zebrafish intestine was analyzed. The efficacy of the samples in improving diarrhea induced by Staphylococcus aureus was evaluated based on the statistical analysis results of this index. The statistical treatment 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.

[0075] According to the above method, medium-chain and long-chain triglycerides and nervonic acid were administered to zebrafish at different dosage ratios, and the efficacy of the two single components in improving diarrhea (intestinal fluorescence intensity) was observed. The administration dosages are shown in Table 2. The administration results are shown in Table 2, Figure 2 、 Figure 3 as shown.

[0076] Table 2 Experimental results of the efficacy evaluation of the composition in improving diarrhea (intestinal fluorescence intensity)

[0077]

[0078] Note: The P value is the significance of the difference in intestinal fluorescence intensity between the medium-chain and long-chain triglyceride group or the nervonic acid group and the model control group.

[0079] From Table 2, Figure 2 、 Figure 3The results showed that, compared with the model control group, the compositions of Examples 1-11 all had the effect of improving diarrhea, specifically manifested as enhanced intestinal fluorescence intensity (P<0.05). Compared with the effects of individual MLCT (concentration 250 μg / mL) and individual nervonic acid (concentration 62.5 μg / mL) at their optimal concentrations, the intestinal fluorescence intensity values of zebrafish in Examples 2-10 were all significantly enhanced, indicating a synergistic effect between MLCT and nervonic acid at a ratio of (3-1500):1; among them, the intestinal fluorescence intensity values of zebrafish in Examples 2-9 with a ratio of MLCT to nervonic acid of (3-800):1 were higher, indicating a more significant synergistic effect between MLCT and nervonic acid at a ratio of (3-800):1; the intestinal fluorescence intensity values of zebrafish in Examples 2-7 with a ratio of MLCT to nervonic acid of (3-150):1 were further increased, indicating a further enhanced synergistic effect between MLCT and nervonic acid at a ratio of (3-150):1; the intestinal fluorescence intensity values of zebrafish in Examples 3-7 with a ratio of MLCT to nervonic acid of (15-150):1 were even further enhanced, indicating a further enhanced synergistic effect between MLCT and nervonic acid at a ratio of (15-150):1.

[0080] Efficacy experiment of Example 3 in inhibiting Staphylococcus aureus

[0081] Wild-type AB strain zebrafish at 4 dpf were randomly selected and placed in 6-well plates, with 30 zebrafish in each well (experimental group). The samples were administered in water (concentrations are shown in Table 3), the positive control vancomycin was at a concentration of 1000 μg / mL, and a normal control group and a model control group were set up simultaneously, with a volume of 3 mL per well. After treatment at 28°C for 24 h. Except for the normal control group, the remaining experimental groups were all administered Staphylococcus aureus in water to establish a zebrafish Staphylococcus aureus infection model. After labeling Staphylococcus aureus with CM-DiI at 28°C for 6 h, 10 zebrafish were randomly selected from each experimental group and placed under a fluorescence microscope for photographing, and data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software to analyze the fluorescence intensity of Staphylococcus aureus in the zebrafish intestine, and the antibacterial efficacy of the samples was evaluated based on the statistical analysis results of this index. The statistical processing results were expressed as mean±SE. Statistical analysis was performed using SPSS 26.0 software, and p<0.05 indicated that the difference was statistically significant.

[0082] The administration doses and results are shown in Table 3.

[0083] Table 3 Results of the efficacy evaluation experiment against Staphylococcus aureus

[0084]

[0085] Note: The P value represents the significance of the difference in the fluorescence intensity of Staphylococcus aureus between the medium and long-chain fatty acid triglyceride group or nervonic acid group, etc., and the model control group.

[0086] As can be seen from Table 3, Figure 4 , Figure 5 the results show that compared with the model control group induced by Staphylococcus aureus, the positive control (vancomycin), monomer medium and long-chain fatty acid triglyceride (250 μg / mL), nervonic acid (62.5 μg / mL), and the compositions of Formulas 1-11 all have significant anti-Staphylococcus aureus effects. Specifically, compared with the model control group, the fluorescence intensity of Staphylococcus aureus in zebrafish becomes weaker (P<0.05). Compared with the monomer medium and long-chain fatty acid triglyceride (250 μg / mL) and nervonic acid (62.5 μg / mL), the anti-Staphylococcus aureus effects of Formulas 2-7 with an MLCT to NA ratio of (3-150):1 are stronger, specifically reflected in the more significant weakening of the fluorescence intensity of Staphylococcus aureus, indicating a synergistic effect; among them, the anti-Staphylococcus aureus effect of Formula 2-6 with an MLCT to NA ratio of (3-100):1 is stronger, indicating a more significant synergistic effect; the anti-Staphylococcus aureus effect of Formula 2-5 with a ratio of (3-66):1 is even further enhanced, indicating a further enhanced synergistic effect.

[0087] Evaluation of the anti-inflammatory effect of the composition in Example 4

[0088] Randomly select 4dpf transgenic neutrophil green fluorescent zebrafish (MPX) in a 6-well plate, with 30 zebrafish in each well (experimental group). Administer the samples in water solution (concentrations are shown in Table 4), the positive control vancomycin at a concentration of 1000 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 24 h, except for the normal control group, the remaining experimental groups are all given Staphylococcus aureus in water solution to establish a zebrafish inflammation model. After treatment at 28°C for 6 h, randomly select 10 zebrafish from each experimental group and place them under a fluorescence microscope for photographing. Use NIS-Elements D 3.20 advanced image processing software to analyze and collect data, and analyze the number of neutrophils in the zebrafish intestine. Evaluate the effect of the samples on the resolution of inflammation induced by Staphylococcus aureus based on the statistical analysis results of this index. The statistical processing results are expressed as mean±SE. Use SPSS26.0 software for statistical analysis, and P<0.05 indicates that the difference is statistically significant.

[0089] According to the above method, set different dosage ratios of medium and long-chain fatty acid triglyceride and nervonic acid to administer to zebrafish to detect their anti-inflammatory effects (the number of neutrophils in the intestine). The dosage and administration results are shown in Table 4.

[0090] Table 4 Experimental results of the anti-inflammatory effect evaluation of the composition

[0091]

[0092] Note: The P value represents the significance of the difference between each experimental group and the model control group.

[0093] Figure 7 Typical graph showing the number of neutrophils in the zebrafish intestine after sample treatment.

[0094] Table 4 Figure 6 and Figure 7 The results show that, compared with the model control group, the compositions of combinations 1 - 9 with the MLCT and nervonic acid ratio of (0.2 - 800):1 have anti-inflammatory effects, manifested as a decrease in the number of intestinal neutrophils (P < 0.05). Compared with the effects of individual MLCT (concentration 250 μg / mL) and individual nervonic acid (concentration 62.5 μg / mL) at their optimal concentrations, the number of neutrophils in the zebrafish intestine in combinations 2 - 9 decreases, indicating a synergistic effect between MLCT and nervonic acid at a ratio of (3 - 800):1; among them, the number of neutrophils in the zebrafish intestine in combinations 2 - 7 is even lower, indicating a more significant synergistic effect between MLCT and nervonic acid at a ratio of (3 - 150):1; the number of neutrophils in the zebrafish intestine in combinations 3 - 6 further decreases, indicating a further enhanced synergistic effect between MLCT and nervonic acid at a ratio of (15 - 100):1; the number of neutrophils in the zebrafish intestine in combinations 3 - 5 even further decreases, indicating an even further enhanced synergistic effect between MLCT and nervonic acid at a ratio of (15 - 66):1.

[0095] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A nutritional composition comprising medium and long chain fatty acid triglycerides and nervonic acid, wherein the mass ratio of the medium and long chain fatty acid triglycerides to the nervonic acid is from 3:1 to 800:

1.

2. The nutritional composition according to claim 1, wherein the mass ratio of the medium and long chain fatty acid triglycerides to the nervonic acid is from 3:1 to 300:

1.

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

1.

4. A product comprising the nutritional composition according to any one of claims 1-3.

5. Use of the nutritional composition according to claim 1 in the preparation of a product having the following use: preventing or alleviating diarrhea, which is diarrhea caused by Staphylococcus aureus.

6. Use of the nutritional composition according to claim 3 in the preparation of a product having the following use: inhibiting Staphylococcus aureus in vivo or in vitro.

7. Use of the nutritional composition according to claim 2 in the preparation of a product having the following use: preventing or treating inflammation, which is intestinal inflammation caused by Staphylococcus aureus.

8. The use according to any one of claims 5-7, wherein the product is a medicine.

9. The use according to claim 8, wherein the dosage form of the medicine is selected from one or more of tablets, granule powders, capsules and liquids.

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