Breast milk oligosaccharide-containing composition for promoting brain metabolism and relieving diarrhea and application of breast milk oligosaccharide-containing composition

Abstract

The invention belongs to the technical field of nutritional compositions, and discloses a breast milk oligosaccharide-containing composition for promoting brain metabolism and relieving diarrhea and application of the breast milk oligosaccharide-containing composition, and the breast milk oligosaccharide-containing composition contains breast milk oligosaccharide consisting of 2 '-fucosyllactose, 3'-sialic acid lactose and 6 '-sialic acid lactose in a specific weight ratio. The composition disclosed by the invention is used for preparing products with the functions of promoting brain development or metabolism and relieving diarrhea. The composition prepared by the invention can be used for preparing formula milk powder, yoghourt and various metabiotics preparations with corresponding effects.

Description

Technical Field

[0001] The present invention belongs to the technical field of nutritional compositions, in particular to a composition containing human milk oligosaccharides for promoting brain metabolism and relieving diarrhea and an application thereof. Background Art

[0002] Human milk oligosaccharides (HMOs) are a class of complex oligosaccharides composed of monosaccharides and their derivatives, sialic acid, and other structural units linked by glycosidic bonds. These include 2'-fucosyllactose (2'-FL), 3'-sialyllactose (3'-SL), and 6'-sialyllactose (6'-SL). Currently, over 150 HMOs have been identified in human milk. HMOs are the third most abundant solid component in human breast milk, after lactose and fat, and have important physiological functions. Because most oligosaccharides in breast milk are indigestible to infants, HMOs can serve as prebiotics in the form of indigestible carbohydrates. They are selectively fermented by specific intestinal microbiota and are crucial for the healthy development of infants in the first year of life.

[0003] Research on the functions of HMOs has made significant progress in recent years, but many unknown areas remain. On the one hand, HMOs can act on multiple aspects such as the immune system, intestinal microbiota, and neurodevelopment, but the specific mechanisms of action and potential effects of oligosaccharides still need further exploration. On the other hand, the demand for different types of HMOs varies at different stages of infant development. Excessive intake of the same type of HMOs at different times will not only lead to poor absorption, but may also place unnecessary burden on the gastrointestinal tract of infants and young children. Therefore, exploring the interaction principles of HMOs is one of the keys to making the compound design of HMOs more targeted.

[0004] The infant and toddler stage is a critical period for brain development and cognitive function formation. Brain development directly affects intelligence, emotions, social skills and future learning ability. Good brain metabolism is crucial for the brain development of infants and toddlers. However, at present, the mechanism of action of fucosyllactose and sialyllactose on brain development and specific metabolic pathways, as well as the effect of further compounding, are still unclear.

[0005] During the growth of infants and young children, key nutrients required for brain development are absorbed through the gastrointestinal tract, enter the bloodstream, and ultimately reach the brain. Therefore, the health and function of the gastrointestinal tract are directly related to the efficiency of nutrient absorption. Infant diarrhea is one of the most common manifestations of gastrointestinal dysfunction. Diarrhea leads to significant water and electrolyte loss, and in severe cases, can lead to dehydration. Beyond these direct consequences, frequent diarrhea can also lead to intestinal dysfunction, impairing nutrient absorption and, in turn, affecting brain development.

[0006] At present, functional nutritional products with HMOs as the main ingredient not only need to solve the problem of single functionality, but also face problems such as how to select specific functional human milk oligosaccharides from nearly 200 HMOs and how to enhance their specific efficacy through compound design. Summary of the Invention

[0007] In order to address the above deficiencies in the prior art, the present invention aims to provide a human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea, and its application in the preparation of dairy products and other foods, so as to achieve the purpose of promoting brain metabolism and relieving diarrhea.

[0008] To achieve the above object, the technical solutions adopted by the present invention are as follows:

[0009] A human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea contains complex human milk oligosaccharides composed of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose in a weight ratio of 0-100:1-100:1-100.

[0010] Furthermore, the weight ratio of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose is 7::0.9:2.1.

[0011] As a limitation of the present invention, it further comprises lactoferrin, and the weight ratio of the lactoferrin to the complex human milk oligosaccharide is 1:0.5-10.

[0012] The present invention also provides the use of two human milk oligosaccharide compositions for promoting brain metabolism and relieving diarrhea in the preparation of products for promoting brain development or metabolism.

[0013] As one of the applications, it is used to increase the content of the brain metabolite lactate and total N-acetylaspartate consisting of N-acetylaspartate and N-acetylaspartylglutamate.

[0014] Another application of the lactoferrin-containing composition is to increase the content of the brain metabolite glutathione and total choline, which consists of phosphocholine and glycerophosphocholine.

[0015] The present invention also provides a use of a human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea in the preparation of a product for relieving diarrhea.

[0016] The present invention also provides a dairy product, the raw materials of which include the above-mentioned human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea.

[0017] The present invention also provides a postbiotic product, which comprises the above-mentioned human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea.

[0018] Due to the adoption of the above technical solution, the present invention has the following beneficial effects compared with the prior art:

[0019] The present invention's human milk oligosaccharide composition, which promotes brain metabolism and alleviates diarrhea, contains a specific ratio of complex human milk oligosaccharides composed of 2'-fucosyllactose, 3'-sialyllactose, and 6'-sialyllactose. These three human milk oligosaccharides synergize to promote brain development, improve cognitive function, and alleviate diarrhea. By combining it with lactoferrin, it provides a more comprehensive nutritional component and achieves synergistic synergy between human milk oligosaccharides and lactoferrin, further enhancing its effects on brain development and gastrointestinal health.

[0020] The human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea of ​​the present invention is used as a raw material to prepare products that promote brain development, local brain metabolism, promote neural development or improve cognitive function, wherein the composite human milk oligosaccharide promotes brain development by regulating special metabolic pathways.

[0021] The human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea of ​​the present invention is used as a raw material for preparing products for promoting brain development, brain metabolism, neural development or improving cognitive function.

[0022] The dairy product with diarrhea relief function of the present invention can promote early digestive tract health and affect overall intestinal comfort. The complex human milk oligosaccharides in its raw materials act on the gastrointestinal tract, promote intestinal development, regulate specific metabolic pathways, and change the intestinal flora environment to reduce the incidence of diarrhea, reduce the frequency of diarrhea or shorten the duration of diarrhea, thereby alleviating diarrhea problems caused by factors such as weaning, and improving the consumer's ability to digest and absorb other nutrients such as protein and lipids in the dairy product raw materials, thereby improving the efficacy of each component and promoting brain development and the healthy development of the body.

[0023] The present invention is suitable for the industrial production of functional foods or health products such as dairy products and postbiotic products containing human milk oligosaccharide compositions. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a graph showing the absolute concentrations of brain metabolites at TE 30 for the Control group, nHMOs group, sHMOs group, and cHMOs group in Example 7 of the present invention;

[0025] Figure 2 This is a graph showing the absolute concentrations of brain metabolites at TE 30 for the Control group, cHMOs group, and cHMOs+Lf group in Example 7 of the present invention;

[0026] Figure 3 This is a graph showing the relative concentrations of brain metabolites in the Control group, cHMOs group, and cHMOs+Lf group in Example 7 of the present invention;

[0027] Figure 4 Graph showing the diarrhea severity test results for the control group, cHMOs group, nHMOs group, and sHMOs group in Example 8 of the present invention, wherein the blue curve represents the control group, the yellow curve represents the cHMOs group, the red curve represents the nHMOs group, and the green curve represents the sHMOs group;

[0028] Figure 5 Graph showing the diarrhea severity test results for the control group, cHMOs group, and cHMOs+Lf group in Example 8 of the present invention, wherein the blue curve represents the control group, the yellow curve represents the cHMOs group, and the purple curve represents the cHMOs+Lf group. DETAILED DESCRIPTION

[0029] The present invention is further described in detail below through specific examples. It should be understood that the described examples are only used to illustrate the present invention and are not intended to limit the present invention.

[0030] Unless otherwise specified, the experimental methods used in the following examples are conventional methods in the art.

[0031] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0032] In the context of the present invention, the term "composition" refers to any composition that provides nutritional benefits to an individual and is safe for human or animal consumption. The composition can be in solid, semi-solid, or liquid form and can contain one or more macronutrients, micronutrients, food additives, water, and the like. For example, the composition can contain the following macronutrients: a protein source, a lipid source, a carbohydrate source, and any combination thereof. In addition, the composition can contain the following micronutrients: vitamins, minerals, fiber, phytochemicals, antioxidants, prebiotics, probiotics, and any combination thereof. The composition can also contain food additives, such as stabilizers or emulsifiers.

[0033] Examples 1 to 6 A human milk oligosaccharide composition for promoting brain metabolism and alleviating diarrhea

[0034] Example 1 is a human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea. It is a complex human milk oligosaccharide composed of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose. The raw materials are: 100 kg of 2'-fucosyllactose, 100 kg of 3'-sialyllactose and 100 kg of 6'-sialyllactose.

[0035] Example 2 is a human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea. It is a complex human milk oligosaccharide composed of 3'-sialyllactose and 6'-sialyllactose. The raw materials are: 10 kg of 3'-sialyllactose and 10 kg of 6'-sialyllactose.

[0036] Example 3 is a human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea. It is a complex human milk oligosaccharide composed of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose. Its raw materials are: 55 kg of 2'-fucosyllactose, 18 kg of 3'-sialyllactose and 35 kg of 6'-sialyllactose.

[0037] Example 4 is a human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea. The composition is composed of a complex human milk oligosaccharide composed of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose and lactoferrin. The raw materials are: 1 kg of lactoferrin, 5 kg of 2'-fucosyllactose, 3 kg of 3'-sialyllactose and 2 kg of 6'-sialyllactose.

[0038] Example 5 is a human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea. It is composed of complex human milk oligosaccharides composed of 3'-sialyllactose and 6'-sialyllactose and lactoferrin. Its raw materials are: 10 kg of lactoferrin, 2.5 kg of 3'-sialyllactose and 2.5 kg of 6'-sialyllactose.

[0039] Example 6 is a human milk oligosaccharide composition that promotes brain metabolism and relieves diarrhea. The composition is composed of a complex human milk oligosaccharide composed of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose, and lactoferrin. The raw materials are: 1 kg of lactoferrin, 3 kg of 2'-fucosyllactose, 1 kg of 3'-sialyllactose and 3 kg of 6'-sialyllactose.

[0040] Example 7: A composition containing human milk oligosaccharides for promoting brain metabolism and alleviating diarrhea

[0041] 1. Experimental principle:

[0042] Proton Magnetic Resonance Spectroscopy (1H-MRS): Proton magnetic resonance spectroscopy is a non-invasive imaging technique used to identify and quantify cellular metabolites and biomarkers in the brain. By analyzing the spectral signal emitted by proton nuclei (1H), MRS analyzes the chemical composition of tissue in specific regions of interest. Brain metabolites detected by MRS include N-acetylaspartate (NAA), creatine (Cr), choline (Cho), inositol (mI), and glutamine / glutamate (Glx). This provides insight into how nutritional intervention with different HMOs affects the concentrations of important neurotransmitters and brain metabolites in piglets, an ideal animal model for human infants.

[0043] 2. Experimental Grouping:

[0044] Healthy, three-day-old, round-headed male piglets (n = 80) from five different strains, weighing 1.3-1.6 kg at birth, were collected from the Grong Grong farm of Pig Improvement Corporation (PIC) in New South Wales (NSW), Australia. The piglets were randomly assigned to five groups (n = 16 each) and fed milk supplemented with formula from 3 to 38 days of age. Some piglets died during the feeding period due to severe diarrhea and fever. The final available data included 14 piglets in the control group, 16 in the cHMOs group, 14 in the cHMOs + Lf group, 15 in the nHMOs group, and 16 in the sHMOs group.

[0045] The formula milk powder for each group is as follows:

[0046] Group 1 (sHMOs): fed with formula containing HMOs of 3′-SL + 6′-SL at a weight ratio of 1:2.5, similar to that in breast milk;

[0047] Group 2 (nHMOs): fed with formula containing only 2′-FL of HMOs;

[0048] Group 3 (cHMOs): fed with formula milk containing HMOs of 2'-FL+3'-SL+6'-SL, with a weight ratio of 7:0.9:2.1;

[0049] Group 4 (cHMOs+Lf): fed with formula milk containing 2'-FL+3'-SL+6'-SL+Lf HMOs (the same dose of HMOs as in Group 3);

[0050] Group 5 (Control): fed the same piglet milk replacer with methylcellulose alone as a control.

[0051] The total dose of HMOs was the same in each group. Based on the safe dose of infant formula (US FDA) and lactoferrin 0.5g / L, the HMOs used in each piglet's milk corresponded to the dose of 1.8g / L in human mature milk.

[0052] Piglets were fed 285 mL / kg / day of milk for the first two weeks of the study. For the remaining weeks, they were fed 230 mL / kg / day of milk at 8:00, 1:00, 18:00, and 22:30, with an additional 50 mL of milk at the last feeding. Piglets were housed in an air-conditioned room at 27°C under a heat lamp. Body weight, milk intake, and health status were recorded daily.

[0053] 3. Specific test contents include:

[0054] Brain development and metabolism exploration experiment: On the 37th and 38th days, the piglet brains were examined using MRS scanning, and tissue samples and blood samples from different regions of the brain were collected from each group of experimental animals that were immediately killed by injection at 38 days of age (equivalent to approximately 10-12 months old human infants) and stored at -80°C for further analysis of brain development, neural development, and local brain metabolism. A 3T magnetic resonance spectrometer was used to acquire brain spectra at an echo time (TE) of 30ms to visualize the differences in 33 metabolite components. Absolute and relative metabolite concentrations (ratios compared to total creatine, choline, and N-acetylaspartate) were determined using TARQUIN software. Using MRS (local brain metabolism), changes in brain metabolism in experimental animals were observed to discover the mechanism of action of the human milk oligosaccharide composition that promotes brain metabolism and alleviates diarrhea in neuroprotection.

[0055] Detection of the mean (±SE) absolute concentrations of 33 brain metabolites: At TE30ms, the mean (±SE) absolute concentrations of 33 brain metabolites in the control group, nHMOs group, sHMOs group, cHMOs group and (cHMOs+Lf) group were measured respectively. The specific metabolites were: alanine (Ala), aspartic acid (Asp), creatine (Cr), γ-aminobutyric acid (GABA), glycerylphosphorylcholine (GPC), d-glucose (Glc), glutamine (Gln), glutathione (Glth), glutamate ( Glu), inositol (Ins), lactate (Lac), lipids (Lip) (09, 13a, 13b, 20), macromolecules (MM) (09, 12, 14, 17, 20), N-acetylaspartic acid (NAA), N-acetylaspartylglutamate (NAAG), phosphocholine (PCh), phosphocreatine (PCr), scyllo-inositol (SI), taurine (Tau), total NAA (TNAA), total choline (TCh), total creatine (TCr), glutamate and glutamine (Glx), total lipids and macromolecules (TLM).

[0056] Among them, the values ​​of lipids and macromolecules refer to the chemical shift values ​​expressed in parts per million (ppm) in the magnetic resonance spectrum of the substance. This reflects the environment of the hydrogen atom (^1H). Lip09 represents the methyl group in the lipid; Lip13a / b represents the methylene group in the fatty acid; Lip20 represents the allyl proton; MM09-20 represents the amino acid side chains in macromolecules (proteins, peptides) with different shifts. TLM usually represents the sum of the signal intensities of various lipid (Lip) and macromolecule (MM) peaks. TLM09 and TLM20 refer to the combined signals of two specific ppm values: 09ppm (mainly related to the terminal methyl group of the lipid); 20ppm (usually related to the allyl-CH2- group in unsaturated lipids and macromolecules).

[0057] 4. Specific test results:

[0058] (1) Absolute concentrations of brain metabolites at TE30 in the four groups:

[0059] The mean (±SE) absolute concentrations of 33 brain metabolites in the control group, nHMOs group, sHMOs group, and cHMOs group were compared. Figure 1 As shown, Figure 1 *P<0.05, general linear model (univariate).

[0060] Among them, lactate is transferred from astrocytes to neurons to match the energy needs of neurons and provide signals to regulate neuronal functions, including excitability, plasticity and memory consolidation. Lactate ensures adequate energy supply, regulates the level of neuronal excitability and regulates adaptive functions to set the "homeostatic tone" of the nervous system (Magistretti and Allaman 2018).

[0061] Depend on Figure 1 The mean absolute concentration of lactate (Lac), a brain metabolite, showed significant overall differences among the four groups. The cHMOs group had the highest absolute concentration of lactate (Lac) (20.72±2.59mM) (P<0.05), followed by the nHMOs group (17.63±1.40mM), the sHMOs group (16.14±1.75mM), and the control group (13.30±0.98mM). The analysis found that only the comparison between the cHMOs group and the control group was statistically significant (P<0.05).

[0062] Furthermore, the cHMOs group had the highest concentrations of most metabolites, including Asp, GABA, Lip, PCh, PCr, SI, Tau, TNAA, TCh, TCr, and TLM. Compared with the cHMOs group, TNAA decreased by 1% in the control group, 5% in the sHMOs group, and 10% in the nHMOs group. GABA increased by 32-40% in the cHMOs group compared with the control, sHMOs, and nHMOs-treated groups.

[0063] The above results show that the cHMOs group using the ratio of the present invention can significantly increase the level of lactic acid (Lac), promote the accumulation of neuroprotective and neuroactive metabolites such as Asp and GABA, regulate brain metabolism, and thus promote neural development and brain development.

[0064] (2) Absolute concentrations of brain metabolites at TE30 in the three groups:

[0065] The mean (±SE) absolute concentrations of 33 brain metabolites in the control group, cHMOs group, and cHMOs+Lf group were compared. Figure 2 , *P<0.05, general linear model (univariate).

[0066] The absolute concentrations of the brain metabolites TNAA and TLM showed significant differences among the three groups (P < 0.05). The mean absolute concentration of TNAA was highest in the cHMOs group (122.32 ± 3.98 mM), followed by the control group (120.98 ± 3.90 mM) and the cHMOs + Lf group (105.54 ± 5.75 mM). Only the cHMOs group and the cHMOs + Lf group showed statistical significance (P < 0.05). TNAA is a combination of NAA and NAAG, a key MRS marker for neuronal cell integrity, neuronal health, viability, and number (Moffett et al., 2007). It promotes energy metabolism in neuronal mitochondria and serves as a source of acetate for fatty acid and steroid synthesis in oligodendrocytes of the nervous system (Ariyannur et al., 2008). NAAG regulates glutamate release and plays a role in neuroprotection and synaptic plasticity (Bubenikova-Valesova et al., 2006).

[0067] On the other hand, the mean absolute concentration of TLM was highest in the cHMOs + Lf group (173.25 ± 29.78 mM), followed by cHMO (146.88 ± 9.93 mM) and the control group (132.52 ± 10.01 mM). Among the groups, only the cHMO + Lf group was statistically significant compared with the control group (P < 0.05). Elevated TLM levels are associated with neurodevelopmental processes and can indicate inflammation, gliosis, or neurodegeneration under pathological conditions (Kreis, 2004).

[0068] The cHMOs group showed the highest concentrations of most metabolites, including Asp, GABA, Glc, Lac, NAAG, PCh, PCr, SI, Tau, TNAA, TCh, and TCr. GABA in the cHMOs group was 18% and 33% higher than in the cHMOs+Lf and control groups, respectively. TNAA decreased by 25% in the control group and 118% in the cHMOs+Lf group, respectively, compared to the cHMOs group. This suggests that the cHMOs group was superior in promoting the metabolism of these substances.

[0069] In the cHMOs+Lf group, the metabolite Ala increased by 78% compared to the control group and by 211% compared to the cHMOs group. The metabolite Cr increased by 6% compared to the control group and by 16% compared to the cHMOs group. The metabolite Glth increased by 10% compared to the control group and by 16% compared to the cHMOs group. The metabolite Lip increased by 46% compared to the control group and by 24% compared to the cHMOs group. The metabolite MM increased by 23% compared to the control group and by 14% compared to the cHMOs group. The metabolite TLM increased by 31% compared to the control group and by 18% compared to the cHMOs group. In other words, the cHMOs+Lf group was superior in promoting the metabolism of these substances.

[0070] The above results show that the compound combination of 2'-FL+3'-SL+6'-SL of the present invention can significantly promote the metabolism of brain metabolites including TNAA and GABA, and increase the concentration of metabolites such as Asp; the compound combination of 2'-FL+3'-SL+6'-SL+Lf can increase the concentration of brain metabolites such as Ala, thereby regulating brain metabolism, and further promoting neural development and brain development.

[0071] (3) Relative concentrations of brain metabolites at TE30 in the three groups:

[0072] The relative concentrations of 15 brain metabolites in the control group, cHMOs group, and cHMOs+Lf group were compared by comparing the absolute concentrations of brain metabolites to total N-acetylaspartate (TNAA) at TE30. The results are shown in Figure 3, *P<0.05, general linear model (univariate).

[0073] Figure 3 At TE 30ms, Glth, TCh, TCr, and TLM20 concentrations were significantly higher than those in the control group (P<0.05). Glth concentration was highest in the cHMOs+Lf group (0.23±0.02mM), followed by the control group (0.18±0.01mM), and finally the cHMOs group (0.17±0.02mM). The analysis revealed that Glth concentration in the cHMOs+Lf group was significantly higher than that in the cHMOs group. The mean relative concentrations of TCh, TCr, and TLM20 were highest in the cHMOs+Lf group (0.29±0.01mM, 0.99±0.04mM, and 2.39±0.22mM, respectively), followed by the cHMOs group (0.25±0.01mM, 0.87±0.03mM, and 1.72±0.12mM, respectively) and the control group (0.23±0.01mM, 0.84±0.03mM, and 1.53±0.11mM, respectively).

[0074] Among them, glutathione (Glth) is a classic antioxidant, playing a role in preventing damage to vital cellular components by reactive oxygen species such as free radicals, peroxides, lipid peroxides, and heavy metals (Pompella et al., 2003). Total choline (TCh) is a combination of phosphocholine (PCh) and glycerophosphocholine (GPC). Choline (Cho) is an essential nutrient and a precursor molecule of the neurotransmitter acetylcholine, involved in many functions, including memory and muscle control (Zeisel, 2006). Choline is also a component of sphingomyelin and phosphatidylcholine, which are components of the myelin sheath (Wang et al., 2019). Total creatine (TCr) is a combination of creatine (Cr) and phosphocreatine (PCr). Creatine (Cr) promotes the recycling of adenosine triphosphate (ATP), the cellular energy currency, primarily found in muscle and brain tissue (Braissant et al., 2011; Brosnan and Brosnan 2016). Increased levels of TLM are associated with neurodevelopmental processes and may indicate inflammation, gliosis, or neurodegeneration under pathological conditions (Kreis, 2004).

[0075] The cHMOs+Lf group showed higher concentrations in most metabolites, including Asp, Ins, PCr, GPC, Glth, TCh, TCr, NAA, and TLM, with prominent neurodevelopmental advantages, while the cHMOs group exhibited higher concentrations in SI.

[0076] The above results show that the combination of 2'-FL+3'-SL+6'-SL and lactoferrin of the present invention can significantly increase the concentration of glutathione, total choline composed of phosphocholine and glycerophosphocholine, thereby regulating brain metabolism and promoting neural development and brain development.

[0077] Example 8: Human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea

[0078] 1. Experimental grouping: Same as Example 7.

[0079] 2. Experimental methods:

[0080] From day 3 to day 35, the fecal scores and diarrhea conditions of the pigs were recorded four times a day, and the average value of the daily data (mean ± SEM) was calculated, and the fecal consistency score was performed to explore the effect of the human milk oligosaccharide composition of the present invention for promoting brain metabolism and relieving diarrhea in improving fecal scores and alleviating the severity of diarrhea.

[0081] The fecal concentration of each piglet was monitored daily using the standard Bristol fecal scoring method, which is suitable for piglets to assess gastrointestinal health. The feces were scored based on visual and physical characteristics as follows:

[0082] 1 point: Sheep dung balls: hard balls (difficult to pass through)

[0083] 2 points: Twist-like: Twist-like, but with uneven surface

[0084] 3 points: Sausage-shaped: Sausage-shaped, but with cracks on the surface

[0085] 4 points: Banana-shaped: like a sausage or snake, with a smooth surface

[0086] 5 points: Marshmallow-like: soft chunks with smooth edges (easy to pass through)

[0087] 6 points: soft and thick: fluffy lumps with rough edges, mushy stool

[0088] 7 points: Liquid: watery, no solid blocks (completely liquid)

[0089] The scores for each piglet were recorded daily by the relevant feeding staff. Each observer was trained and calibrated to ensure consistency in scoring and adherence to hygiene protocols to minimize external influences on fecal quality. Higher scores were considered more severe diarrhea.

[0090] Among them, the statistical results were analyzed using two-way analysis of variance.

[0091] 3. Experimental Results

[0092] (1) Severity of diarrhea (comparison of 4 groups):

[0093] Comparing the results of the control group represented by the blue curve, the cHMOs group represented by the yellow curve, the nHMOs group represented by the red curve, and the sHMOs group represented by the green curve, as shown in Figure 4 In terms of diarrhea severity, the cHMOs and sHMOs groups had significantly lower diarrhea severity than the control group. The control group vs. sHMOs group (P = 0.0401) and the control group vs. cHMOs group (P = 0.0343) showed that supplementing with the HMOs combination significantly improved diarrhea in piglets.

[0094] (2) Severity of diarrhea (comparison among three groups):

[0095] Comparing the results of the control group represented by the blue curve, the cHMOs group represented by the yellow curve, and the cHMOs+Lf group represented by the purple curve, Figure 5 In terms of diarrhea severity, the cHMOs group had significantly lower diarrhea severity than the control group. The cHMOs+Lf group also had significantly lower diarrhea severity than the control group, with the difference being even more significant. The control group vs. the cHMOs+Lf group had a P=0.0003; the control group vs. the cHMOs group had a P=0.0098.

[0096] The above results show that supplementing the HMOs-containing composition of the present invention can significantly improve diarrhea in piglets and has a synergistic effect with lactoferrin.

[0097] Example 9 Application of a human milk oligosaccharide composition for promoting brain metabolism and alleviating diarrhea

[0098] The human milk oligosaccharide compositions of Examples 1 to 6 that promote brain metabolism and relieve diarrhea are respectively used to prepare dairy products that promote brain development, local brain metabolism or promote neural development, and are fed to infants to achieve the application in the preparation of products that promote brain development, neural development or local brain metabolism.

[0099] The human milk oligosaccharide compositions of Examples 1 to 6, which promote brain metabolism and alleviate diarrhea, were used to prepare postbiotic products for promoting intestinal development or alleviating diarrhea, and then fed to infants to achieve their use in the preparation of products for promoting intestinal development or alleviating diarrhea. Animal experiments have confirmed that consumption of the postbiotic products of the present invention can promote intestinal development, reduce the incidence of diarrhea, reduce the frequency of diarrhea, or shorten the duration of diarrhea.

[0100] It should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art may still modify the technical solutions described in the above embodiments or replace some of the technical features therein with equivalents. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the scope of protection of the claims of the present invention.

Claims

1. A human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea, characterized in that: The invention contains composite human milk oligosaccharide, which is composed of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose in a weight ratio of 0-100:1-100:1-100.

2. The human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea according to claim 1, characterized in that: The weight ratio of 2'-fucosyllactose, 3'-sialyllactose and 6'-sialyllactose is 7:0.9:2.

1.

3. A human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea according to claim 1 or 2, characterized in that: It also contains lactoferrin, and the weight ratio of the lactoferrin to the complex human milk oligosaccharide is 1:0.5-10.

4. Use of the human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea according to claim 2 in the preparation of a product for promoting brain development or metabolism.

5. Use of the human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea as claimed in claim 3 in the preparation of products for promoting brain development or metabolism.

6. The use according to claim 4, characterized in that Used to increase the content of the brain metabolite lactate and total N-acetylaspartate, which consists of N-acetylaspartate and N-acetylaspartylglutamate.

7. The use according to claim 5, characterized in that Used to increase the content of the brain metabolite glutathione and total choline, which is composed of phosphocholine and glycerophosphocholine.

8. Use of the human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea according to claim 3 in the preparation of a product for relieving diarrhea.

9. A dairy product, characterized in that The raw material of the dairy product comprises the human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea according to any one of claims 1 to 3.

10. A postbiotic product, characterized in that The postbiotic product comprises a human milk oligosaccharide composition for promoting brain metabolism and relieving diarrhea according to any one of claims 1 to 3.

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