Functional nutritional composition for use in assisting brain immune regulation

Abstract

The present invention belongs to the field of functional substance research, and specifically relates to the use of functional nutritional compositions that contribute to brain immunoregulation. The functional nutritional composition of the present invention comprises the following essential ingredients: 3-fucosyllactose and osteopontin, and in the functional nutritional composition, the mass ratio of 3-fucosyllactose to osteopontin is (0.05-36):1. The present invention has found that the combination of 3-fucosyllactose and osteopontin, especially in a certain proportion, has a synergistic effect of contributing to brain immunoregulation, especially regulating the polarization trend of microglia.

Description

Technical Field

[0001] The present invention belongs to the research field of functional substances, and specifically relates to the use of a functional nutritional composition in aiding brain immunoregulation, and more specifically relates to the use of a functional nutritional composition containing 3-fucosyllactose and osteopontin in aiding brain immunoregulation and assisting in improving the health of the central nervous system. Background Art

[0002] Glial cells are widely involved in the physiological functions of the central nervous system (CNS), and play an important role in normal physiological processes such as synaptic transmission, neuroplasticity, regulation of neurons and local microenvironment. Among them, microglia are resident immune cells of the central nervous system, accounting for about 10% of all brain cells. They participate in the removal of pathogens, phagocytize dead or apoptotic cells, and promote or inhibit the occurrence of inflammation to maintain brain health. Under physiological conditions, microglial cell bodies maintain slender protrusions and continuously detect changes in the brain's internal environment. Once stimulated, microglia are instantly activated, the number surges, and the cell body enlarges, and the protrusions decrease and become shorter. When the CNS is adversely stimulated, microglia are activated and differentiated into two polarized phenotypes, which play different functions. One type of microglia is continuously activated and transformed into the M1 phenotype with proinflammatory effects, which may cause neuroinflammation. In severe cases, it can cause neuronal damage, loss of synaptic function, and affect the local microenvironment of the brain. Another type of microglia transforms into the M2 phenotype with anti-inflammatory effects, increasing the secretion of brain-derived neurotrophic factor, which can promote the healing of damage caused by chronic stress and play a neuroprotective role.

[0003] Osteopontin (OPN) is a highly glycosylated and phosphorylated multifunctional acidic protein present in all vertebrates, which contains an arginine-glycine-aspartic acid sequence. Osteopontin is involved in a variety of biological processes, such as activation and regulation of the immune system, biomineralization, growth and development of the intestine and brain, etc. Many studies have also shown that osteopontin plays an important role in organisms, especially in immune activation, bone damage repair, angiogenesis, bone remodeling, and promoting brain development. Osteopontin was originally discovered in bones. It is expressed in many different types of cells and is therefore present in most tissues and physiological secretions. The mass concentration in human milk is relatively high. Osteopontin has the highest concentration in milk, and it is believed to initiate and regulate the development, immunity, and physiological processes of infants who drink milk.

[0004] According to the different structural types, human milk oligosaccharides (HMOs) are mainly divided into neutral fucosylated HMOs (such as 2'-fucosyllactose (2'-FL) and 3-fucosyllactose (3-FL)), neutral non-fucosylated HMOs (such as lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT)) and acidic sialylated HMOs (such as 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL)). Compared with the intensively studied 2'-FL, 3-FL has received less attention in almost all aspects. However, 3-FL is naturally present in breast milk and increases with lactation, playing an irreplaceable role in human milk and providing functional properties to newborns. Studies have shown that 3-FL is believed to reduce the risk of intestinal microbial flora imbalance caused by harmful bacteria and selectively stimulate the growth of beneficial bifidobacteria to support overall intestinal health.

[0005] At present, there have been studies on the physiological functions of osteopontin, human milk oligosaccharides and compositions containing the two, such as:

[0006] Reference 1 found that intranasal administration of OPN can improve neurological dysfunction and brain edema in rats with subarachnoid hemorrhage (SAH), inhibit the expression of CD86, iNOS, IL-1β, and IL-6 in the brain, and promote the expression of CD206, Arg-1, IL-10, and IL-13. OPN reduces the inflammatory response after SAH by inhibiting the polarization of microglia M1.

[0007] Reference 2 investigated the association of HMOs with MRI indices of tissue microstructure and regional cerebral blood flow (rCBF) in infants. In mother-infant pairs (N=20) at 1 month postpartum, 2'-FL, 3-FL, 3'-SL, and 6'-SL concentrations in breast milk were analyzed. The data showed that fucosylated and sialylated HMOs had different associations with tissue microstructure and rCBF indices, suggesting that 2'-FL, 3-FL, and 3'-SL may play a special role in early brain maturation.

[0008] Reference 3 discloses a nutritional composition containing a therapeutically effective amount of 2-fucosyllactose, which is used for treating, preventing or alleviating neuroinflammation, and is also used for treating cognitive dysfunction, preventing cognitive impairment, or reducing cognitive damage, etc.

[0009] Reference 4 discloses a nutritional composition for promoting neurodevelopment, the composition comprising 2'-fucosyllactose and osteopontin. Studies have found that the nutritional composition has a significant synergistic effect in promoting neurodevelopment such as neuronal maturation, synaptogenesis and myelination, especially in promoting the proliferation, maturation and differentiation of oligodendrocyte precursor cells (OPCs) into mature oligodendrocytes (OLs) and / or the myelination characteristics of OLs.

[0010] References 5 and 6 both disclose a nutritional composition comprising 2'-fucosyllactose and osteopontin. Studies have found that such a nutritional composition can improve the brain development and intelligence of infants and young children, especially promote neural development such as promoting neuronal maturation, synapse formation and / or myelination.

[0011] Reference 7 discloses a composition comprising osteopontin and at least one HMO selected from 2'-fucosyllactose, 3-fucosyllactose, difucosyllactose, etc. These compositions can be used to treat and / or prevent viral infections in mammals, such as humans, especially infants, toddlers and preschoolers.

[0012] References:

[0013] Reference 1: Chen Ligang, Zou Zheng, Hao Guangzhi, et al. Osteopontin inhibits the polarization of microglia to M1 in rats with subarachnoid hemorrhage[J]. Journal of Neuroanatomy, 2024, 40(03): 373-377.

[0014] Cited literature 2: Berger PK, Bansal R, Sawardekar S, Yonemitsu C, Furst A, HampsonHE, Schmidt KA, Alderete TL, Bode L, Goran MI, Peterson BS. Associations of HumanMilk Oligosaccharides with Infant Brain Tissue Organization and RegionalBlood Flow at 1Month of Age.Nutrients.2022Sep16;14(18):3820.

[0015] Reference 3: WO2015100091A1;

[0016] Reference 4: CN114586983A;

[0017] Reference 5: CN115413789A;

[0018] Reference 6: CN115316660A;

[0019] Reference 7: CN118632706A. Summary of the invention

[0020] Problem that the invention aims to solve

[0021] At present, the existing technology for the study of fucosyllactose mainly focuses on 2'-fucosyllactose, while there are fewer studies on 3-fucosyllactose. For osteopontin, although the existing technology studies have found that nasal administration of OPN can inhibit the polarization of M1 microglia in SAH rats and thus inhibit the inflammatory response, this does not mean that it can play the same role in combination with any substance or under any conditions.

[0022] During the research process, the present invention unexpectedly discovered that the combination of 3-fucosyllactose and osteopontin has a synergistic effect in helping to regulate brain immunity and assist in improving the health of the central nervous system, especially in regulating the polarization of microglia and improving neuroinflammation. Especially when 3-fucosyllactose and osteopontin are combined in a certain ratio, the effect is better.

[0023] Therefore, the object of the present invention is to provide a functional nutritional composition comprising 3-fucosyllactose and osteopontin in a specific ratio for use in preparing a food that helps brain immunoregulation or assists in improving the health of the central nervous system.

[0024] Solutions for solving problems

[0025] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0026] [1]. Use of a functional nutritional composition in preparing a food that helps brain immune regulation, wherein the functional nutritional composition comprises the following essential ingredients: 3-fucosyllactose and osteopontin, and in the functional nutritional composition, the mass ratio of 3-fucosyllactose to osteopontin is (0.05-36):1.

[0027] [2] The use according to [1], wherein, in the functional nutritional composition, the total mass of 3-fucosyllactose and osteopontin accounts for more than 60% of the total dry weight of the functional nutritional composition.

[0028] [3] The use according to [1] or [2], wherein the contribution to brain immune regulation includes at least one of inhibiting the polarization of microglia to the M1 phenotype, promoting the polarization of microglia to the M2 phenotype, and inducing the transformation of microglia from the M1 phenotype to the M2 phenotype.

[0029] [4] The use according to any one of [1] to [3], wherein the effect of helping brain immune regulation includes at least one of increasing the amount of IL-10 released by microglia, reducing the amount of TNF-α released by microglia, reducing the amount of IL-1β released by microglia, and reducing the amount of NO released by microglia.

[0030] [5]. The use according to any one of [1] to [4], wherein the food is an oral preparation, and the oral preparation includes any one or more of tablets, pills, granules, powders, capsules and oral liquids.

[0031] [6]. The use according to any one of [1] to [4], wherein the food is a dairy product, a candy, a beverage or a baked product.

[0032] [7]. The use according to [6], wherein the dairy product comprises at least one of fermented milk, cheese and milk powder; the candy comprises at least one of hard candy, jelly candy, crisp candy, compressed candy and aerated candy; the beverage comprises at least one of carbonated beverage, tea beverage, coffee beverage, fruit and vegetable juice beverage and lactic acid bacteria beverage; the baked food comprises at least one of bread, cake and biscuits.

[0033] [8]. Use of a functional nutritional composition in the preparation of a food that assists in improving the health of the central nervous system, wherein the functional nutritional composition comprises the following essential ingredients: 3-fucosyllactose and osteopontin, and in the functional nutritional composition, the mass ratio of 3-fucosyllactose to osteopontin is (0.05-36):1.

[0034] [9]. The use according to [8], wherein the auxiliary improvement of central nervous system health includes improving neuroinflammation for non-therapeutic purposes.

[0035]

[10] . The use according to [8] or [9], wherein, in the functional nutritional composition, the total mass of 3-fucosyllactose and osteopontin accounts for more than 60% of the total dry weight of the functional nutritional composition.

[0036] Effects of the Invention

[0037] The present invention has found that the combination of 3-fucosyllactose and osteopontin, especially in a certain proportion, has a synergistic effect on brain immunoregulation and assisting in improving the health of the central nervous system, such as synergistic regulation of microglia polarization and improvement of neuroinflammation. Experimental data show that the trend of BV2 cells to differentiate into M1 phenotype microglia is reduced when 3-fucosyllactose and osteopontin are treated, while the trend of differentiation into M2 phenotype microglia is increased, and the amount of pro-inflammatory factors generated is also significantly reduced. In addition, 3-fucosyllactose and osteopontin have a good synergistic effect, which can play an effective role in helping brain immunoregulation and assisting in improving the health of the central nervous system. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 : Schematic diagram of the experimental process of studying the regulatory effect of functional nutrients on microglia in the present invention.

[0039] Figure 2A : Western blot detection results of the effects of functional nutrients on the TLR4 / NF-κB signaling pathway of BV2 cells stimulated by LPS; among them, group 1 was the LPS model group, group 2 was the blank control group, group 3 was the OPN high-dose group, group 4 was the 3-FL high-dose group, group 5 was the OPN high-dose + 3-FL low-dose group (OPN:3-FL was 1:0.1), and group 6 was the OPN low-dose + 3-FL high-dose group (OPN:3-FL was 1:30).

[0040] Figure 2B : Western blot analysis results of the effects of functional nutrients on TLR4 protein expression in the TLR4 / NF-κB signaling pathway of BV2 cells stimulated by LPS; Group 1 was the LPS model group, Group 2 was the blank control group, Group 3 was the OPN high-dose group, Group 4 was the 3-FL high-dose group, Group 5 was the OPN high-dose + 3-FL low-dose group (OPN:3-FL was 1:0.1), Group 6 was the OPN low-dose + 3-FL high-dose group (OPN:3-FL was 1:30), *P<0.05, ****P<0.0001.

[0041] Figure 2C: Western blot analysis results of the effects of functional nutrients on the expression of NF-κB p65 protein in the TLR4 / NF-κB signaling pathway of BV2 cells stimulated by LPS; Group 1 was the LPS model group, Group 2 was the blank control group, Group 3 was the OPN high-dose group, Group 4 was the 3-FL high-dose group, Group 5 was the OPN high-dose + 3-FL low-dose group (OPN:3-FL was 1:0.1), Group 6 was the OPN low-dose + 3-FL high-dose group (OPN:3-FL was 1:30), *P<0.05, ***P<0.001, ****P<0.0001.

[0042] Figure 2D : Western blot analysis results of the effects of functional nutrients on the expression of p-NF-κB p65 protein in the TLR4 / NF-κB signaling pathway of BV2 cells stimulated by LPS; Group 1 was the LPS model group, Group 2 was the blank control group, Group 3 was the OPN high-dose group, Group 4 was the 3-FL high-dose group, Group 5 was the OPN high-dose + 3-FL low-dose group (OPN:3-FL was 1:0.1), Group 6 was the OPN low-dose + 3-FL high-dose group (OPN:3-FL was 1:30), **P<0.01, ***P<0.001, ****P<0.0001.

[0043] Figure 2E : Western blot analysis results of the effects of functional nutrients on the expression of IKB-α protein in the TLR4 / NF-κB signaling pathway of BV2 cells stimulated by LPS; Group 1 was the LPS model group, Group 2 was the blank control group, Group 3 was the OPN high-dose group, Group 4 was the 3-FL high-dose group, Group 5 was the OPN high-dose + 3-FL low-dose group (OPN:3-FL was 1:0.1), Group 6 was the OPN low-dose + 3-FL high-dose group (OPN:3-FL was 1:30), **P<0.01, ***P<0.001, ****P<0.0001.

[0044] Figure 2F : Western blot analysis results of the effects of functional nutrients on the expression of p-IKB-α protein in the TLR4 / NF-κB signaling pathway of BV2 cells stimulated by LPS; Group 1 was the LPS model group, Group 2 was the blank control group, Group 3 was the OPN high-dose group, Group 4 was the 3-FL high-dose group, Group 5 was the OPN high-dose + 3-FL low-dose group (OPN:3-FL was 1:0.1), Group 6 was the OPN low-dose + 3-FL high-dose group (OPN:3-FL was 1:30), ***P<0.001, ****P<0.0001. DETAILED DESCRIPTION

[0045] The following describes the embodiments of the present invention, but the present invention is not limited thereto. Various changes can be made within the scope of the present invention, and the embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention.

[0046] In the present invention, the term "a" or "an" or "the" may mean "one", and may also mean "one or more", "at least one" and "one or more than one".

[0047] In the present invention, a numerical range expressed using “value A to value B”, “value A - value B”, “value A or more” or “value A or less” means a range including the endpoints A and B.

[0048] In the present invention, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances where the event occurs and instances where it does not occur.

[0049] In the present invention, the word "may" includes both performing a certain process and not performing a certain process.

[0050] In the present invention, the terms "comprising", "having", "including" or "containing" may mean inclusive or open-ended, and do not exclude additional, uncited elements or method steps. At the same time, "comprising", "having", "including" or "containing" may also mean closed, excluding additional, uncited elements or method steps.

[0051] In the present invention, the term "about" is used to define the numerical range and parameters of the present invention are approximate values, and the specific relevant values ​​are presented as accurately as possible. Unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in the present invention are modified by "about". Here, "about" usually means that the actual value is within ±5%, ±3%, ±1% or ±0.5% of a particular value or range.

[0052] Unless otherwise defined, other 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.

[0053] The present invention is mainly completed based on the following insights:

[0054] Lipopolysaccharide (LPS) is the main component of the outer membrane of Gram-negative bacteria. Studies have shown that LPS can enter the brain through the blood-brain barrier to activate microglia and induce neuroinflammatory reactions, induce the release of proinflammatory cytokines, and cause memory impairment, etc. The present invention uses microglia stimulated by LPS as a microglia activation model to study the regulatory effects of different types and concentrations of functional nutrients on microglia, and explore functional nutrients (such as LPS) that are safe (non-cytotoxic) and have obvious effects on brain immune regulation and maintaining the health of the nervous system. Figure 1 The present invention has found through research that the combination of 3-fucosyllactose and osteopontin can be used to intervene in microglia stimulated by LPS, thereby inhibiting the polarization of microglia to M1 phenotype, promoting the polarization of microglia to M2 phenotype, inducing the transformation of microglia from M1 phenotype to M2 phenotype, and increasing the amount of IL-10 released by microglia, and reducing the amount of TNF-α, IL-1β and NO released by microglia.

[0055] I. Functional Nutritional Compositions

[0056] The present invention provides a functional nutritional composition, which comprises the following essential ingredients: 3-fucosyllactose and osteopontin, and in the functional nutritional composition, the mass ratio of 3-fucosyllactose to osteopontin is (0.05-36):1.

[0057] In some embodiments, the 3-fucosyllactose and osteopontin are the main effective ingredients in the functional nutritional composition; that is, the functional nutritional composition mainly relies on the 3-fucosyllactose and osteopontin it contains to exert specific physiologically active functions, such as helping brain immune regulation.

[0058] In some embodiments, in the functional nutritional composition, the total mass of the two substances, 3-fucosyllactose and osteopontin, accounts for more than 60% of the total dry weight of the functional nutritional composition, for example, it can be 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% or 95%, etc., preferably more than 85%, more preferably more than 90%. In some embodiments, in addition to 3-fucosyllactose and osteopontin, the functional nutritional composition can also contain a small amount of other types of carbohydrate substances, fat substances and / or protein substances, such as lactose.

[0059] In some embodiments, the functional nutritional composition consists of the 3-fucosyllactose and osteopontin.

[0060] At present, the production methods of human milk oligosaccharides mainly include chemical synthesis, enzymatic synthesis and bioengineering synthesis. At the same time, it can also be indirectly introduced from various existing milk components.

[0061] It should be noted that the use of human milk oligosaccharides should comply with local laws and regulations. In some cases, human milk oligosaccharides can be directly introduced into food compositions as a separate raw material, where permitted by laws and regulations; in other cases, human milk oligosaccharides can be indirectly introduced into compositions or foods by adding milk raw materials that meet the requirements, where permitted by laws and regulations.

[0062] The present invention does not particularly limit the source of the 3-fucosyllactose, for example, it can be brought in from different animal milk raw materials (such as cow's milk, milk powder, etc.), separated and extracted from natural substances, obtained by chemical synthesis, or obtained by microbial fermentation and the like. In order to facilitate actual production applications, commercial 3-fucosyllactose products can also be used directly. In the present invention, "animal milk" is used to refer to the liquid obtained from the mammary gland of a mammal in the lactation period. The term "animal milk" should be interpreted broadly and covers both raw milk (i.e., liquid obtained directly from the mammary gland) and standardized dairy products.

[0063] The present invention does not specifically limit the source of the osteopontin, for example, it can be extracted from osteopontin-rich cow's milk. In order to facilitate actual production and application, commercial osteopontin products, such as protein powder containing osteopontin and whey powder containing osteopontin, can also be directly used.

[0064] In some embodiments, in the functional nutritional composition, the mass ratio of 3-fucosyllactose to osteopontin is (0.05-36):1; for example, it can be 0.05:1, 0.06:1, 0.07:1, 0.08:1, 0.09:1, 0.1:1, 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1 :1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1 or 36:1, etc.; preferably (0.1-30):1; more preferably (0.1-20):1; further preferably (0.1-10):1; even more preferably (0.5-10):1.

[0065] The present invention finds that when the mass ratio of 3-fucosyllactose to osteopontin is within the above range, the two have a synergistic effect of helping to regulate brain immunity and assist in improving the health of the central nervous system, including regulating the polarization of microglia and improving neuroinflammation.

[0066] II. Use of functional nutritional composition in preparing food

[0067] The present invention provides use of the functional nutritional composition in preparing food.

[0068] In some embodiments, the functional nutritional composition is added to or used in the food so that the mass ratio of 3-fucosyllactose to osteopontin in the food is (0.05-36):1.

[0069] The present invention has no particular restrictions on the absolute content of 3-fucosyllactose and osteopontin in food, as long as the requirements of relevant laws and regulations are met. In some embodiments, the content of 3-fucosyllactose in food is 0.001% to 4%, preferably 0.01% to 2%; the content of osteopontin in food is 0.005% to 0.5%, preferably 0.01% to 0.2%.

[0070] In addition to the functional nutritional composition and the 3-fucosyllactose and osteopontin therein, depending on the type of food and the final needs of the applicable objects, in some embodiments, the food described in the present invention also contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional added ingredients and any acceptable excipients.

[0071] As for the ingredients of plant products, examples include fruits such as figs, pomegranates, kiwis, tangerines, oranges, pineapples, strawberries, apples, bananas, grapes, pears, cherries, blueberries, blackberries, black currants, cranberries, raspberries, melons, emblica chinensis and bilberry or their extracts; vegetables such as onions, cucumbers, tomatoes, cauliflowers, carrots, spinach, kale, Brussels sprouts, garlic, basil, oregano or their extracts; grains such as rice (indica rice, japonica rice, glutinous rice), wheat (wheat, barley, oats, rye), corn, sorghum, millet, sorghum, yellow rice, buckwheat, soybeans, broad beans, peas, mung beans, red beans, kidney beans or their extracts; nuts such as walnuts, pistachios, cashews, hazelnuts, almonds, apricots, pine nuts, peanuts, melon seeds, chestnuts, macadamia nuts, ginkgo nuts or their extracts; coffee or its extracts; and some botanical Chinese medicinal materials or their extracts that are both medicine and food.

[0072] As for animal dairy ingredients, examples include fresh milk from mammals such as cows, sheep, and camels, as well as reprocessed dairy products such as whole milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, and hydrolyzed whey protein powder.

[0073] Examples of animal meat ingredients include meat ingredients from pigs, cattle, sheep, aquatic animals or poultry.

[0074] Functional added ingredients include vitamin supplements, mineral supplements, nucleotide supplements, dietary fiber, functional polyunsaturated fatty acid supplements, and the like.

[0075] As for any acceptable excipients, examples include solvents, antioxidants, antibacterial agents, thickeners, diluents, cosolvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweeteners, edible flavors, edible colorings, etc.

[0076] The present invention has no particular limitation on the types of food, and examples thereof include dairy products, candies, beverages or baked goods. For dairy products, examples include fermented milk, cheese and milk powder. For candies, examples include hard candies, crisp candies, gel candies, compressed candies and aerated candies. For beverages, examples include carbonated beverages, tea beverages, coffee beverages, fruit and vegetable juice beverages and lactic acid bacteria beverages. For baked goods, examples include bread, cakes and biscuits. In addition, the food of the present invention may also be a health food, such as various types of oral preparations, including tablets, pills, granules, powders, capsules and oral liquids.

[0077] The food of the present invention has the function of helping to regulate brain immunity and assisting in improving the health of the central nervous system. The function of helping to regulate brain immunity and assisting in improving the health of the central nervous system of the present invention is not aimed at preventing and treating diseases.

[0078] In some specific embodiments, the helping to brain immune regulation includes inhibiting the polarization of microglia to M1 phenotype, promoting the polarization of microglia to M2 phenotype, and inducing the transformation of microglia from M1 phenotype to M2 phenotype. In some specific embodiments, the helping to brain immune regulation includes inhibiting the polarization of microglia to M1 phenotype, promoting the polarization of microglia to M2 phenotype, and inducing the transformation of microglia from M1 phenotype to M2 phenotype. In some specific embodiments, the helping to brain immune regulation includes increasing the amount of IL-10 released by microglia and reducing the amount of TNF-α, IL-1β and / or NO released by microglia. In some more specific embodiments, the helping to brain immune regulation includes increasing the amount of IL-10 released by microglia and reducing the amount of TNF-α, IL-1β and / or NO released by microglia. In some specific embodiments, the helping to regulate brain immunity comprises increasing the amount of IL-10 released by microglia and reducing the amount of TNF-α, IL-1β and NO released by microglia.

[0079] In some embodiments, the aiding in improving the health of the central nervous system includes improving neuroinflammation, and such neuroinflammation does not reach disease levels, that is, the present invention provides the use of the functional nutritional composition in preparing a food for improving neuroinflammation. In some embodiments, the health of the central nervous system is affected by neuroinflammation.

[0080] In some embodiments, the improvement of neuroinflammation is achieved by affecting the TLR4 / NF-κB signaling pathway in microglia. In some specific embodiments, the effect on the TLR4 / NF-κB signaling pathway in microglia includes inhibiting the expression of TLR4, inhibiting the expression of NF-κB p65, inhibiting the phosphorylation of NF-κB p65, increasing the expression of IKB-α and inhibiting the phosphorylation of IKB-α.

[0081] Example

[0082] The embodiments of the present invention will be described in detail below in conjunction with examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out under normal conditions or conditions recommended by the manufacturer. Unless otherwise specified, the materials or instruments used are all commercially available conventional products.

[0083] 1. Materials and Methods

[0084] 1.1 Raw materials

[0085] Osteopontin (OPN): Arla Foods Ingredients Group P / S, Lacprodan OPN-10;

[0086] 3-Fucosyllactose (3-FL): DSM, GlyCare 3-FL 9001.

[0087] 1.2 Cells

[0088] Mouse microglial cell line BV2 cells were purchased from Shanghai Enzyme Research Biotechnology (CC-Y2022).

[0089] 1.3 Reagents

[0090] Table 1 Main reagents for the experiment

[0091]

[0092] 1.4 Instruments

[0093] Table 2 Main instruments and equipment for the experiment

[0094] serial number Instrument Name factory 1 VD-1320 clean workbench Beijing Donglian Har Instrument Manufacturing Company 2 Carbon dioxide incubator Thermo Fisher Scientific China Co., Ltd. 3 Motic Microscope Harbin Wayne Trading Co., Ltd. 4 Cell Imaging Microplate Reader Meigu Molecular Instruments Co., Ltd. 5 Constant temperature water bath Quanzhou Yiguan Technology Co., Ltd. 6 Multifunctional ELISA reader Dynatech Corporation 7 Centrifuge Thermo Fisher Scientific China Co., Ltd. 8 Fluorescence quantitative PCR instrument Thermo Fisher Scientific China Co., Ltd. 9 Water bath Quanzhou Yiguan Technology Co., Ltd. 10 Nucleic acid protein analyzer Thermo Fisher Scientific China Co., Ltd. 11 Ordinary PCR instrument Harbin Wayne Trading Co., Ltd. 12 Multifunctional imaging system Wuhan Sewell Biotechnology Co., Ltd. 13 Transfer electrophoresis instrument Wuhan Sewell Biotechnology Co., Ltd.

[0095] 1.5 Cell culture

[0096] Mouse microglial cell line (BV2) was cultured in DMEM high-glucose medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37° C. in a 5% CO 2 incubator.

[0097] 1.6 Nutrient cytotoxicity assay

[0098] The CCK-8 kit detects the toxicity of nutrients to cells. The working principle of the CCK-8 kit is to use the dehydrogenase in the cell mitochondria to reduce 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonate benzene)-2H-tetrazole monosodium salt to yellow formazan in the presence of the electron carrier 1-methoxy-5-methylphenazine dimethyl sulfate. Because only mitochondria in living cells can produce dehydrogenase, the production of formazan is directly proportional to the number of living cells, and the OD value is detected at 450nm on the microplate reader.

[0099] BV2 cells were seeded into 96-well plates at 4 × 10 4 Cells, 200 μL / well, 3 replicates per group. Place in an incubator and culture for 12 hours. After the cells are completely attached, add the prepared culture medium of each nutrient to the 96-well plate. After 12 hours of culture, add 20 μL CCK-8 solution (protected from light) to each well and continue incubating in the cell culture incubator for 0.5-1 hour. Use an enzyme marker to measure the absorbance at 450nm, and then use the formula: cell survival rate = [(absorbance of experimental well-absorbance of blank well) / (absorbance of control well-absorbance of blank well)] × 100% (wherein the experimental well contains cells, culture medium, nutrients and CCK-8 solution; the blank well contains culture medium and CCK-8 solution, but does not contain cells and nutrients; the control well contains cells, culture medium and CCK-8 solution, but does not contain nutrients), and finally calculate the cell survival rate. The CCK8 method was used to detect the viability of BV2 microglia.

[0100] 1.7 Establishment of microglial activation model and experimental grouping

[0101] Mouse microglial cell line (BV2) was cultured in a 5% CO2 incubator (37°C, saturated humidity) using high-glucose DMEM medium (containing 10% fetal bovine serum). BV2 cells in logarithmic growth phase were selected, and cells in logarithmic growth phase and reaching 90% confluence were digested with trypsin. The cell concentration was adjusted, and the cells were inoculated in a suitable cell culture dish, and then cultured for another 24 hours before the cells were grouped. 4 The number of cells / mL was inoculated in a 96-well plate, 200 μL / well, and 3 replicates were added to each group. The experiment was divided into 17 groups, namely blank control group, LPS model group, and 15 experimental groups. (1) Control group: conventional cell culture was used; (2) LPS model group: LPS (1 μg / mL) was used to intervene BV2 cells for 24 hours to establish a microglial activation model; (3) Experimental group: LPS stimulated BV2 cells for 24 hours, and each nutrient was added for intervention for 12 hours.

[0102] Table 3 Experimental groups and doses

[0103] Corresponding experiment Dose Group Description LPSμg / mL OPNμg / mL 3-FLμg / mL OPN:3-FL Blank control Blank control group - - - Positive Control LPS model group 1 - - Experiment 1 OPN high dose group 1 500 - Experiment 2 OPN medium dose group 1 250 - Experiment 3 OPN low dose group 1 50 - Experiment 4 3-FL high dose group 1 - 1500 Experiment 5 3-FL medium dose group 1 - 1000 Experiment 6 3-FL medium and low dose group 1 - 250 Experiment 7 3-FL low dose group 1 - 50 Experiment 8 OPN high + 3-FL high 1 500 1500 1:3 Experiment 9 OPN High + 3-FL Medium 1 500 1000 1:2 Experiment 10 OPN high + 3-FL medium low 1 500 250 1:0.5 Experiment 11 OPN high + 3-FL low 1 500 50 1:0.1 Experiment 12 OPN Medium + 3-FL High 1 250 1500 1:6 Experiment 13 OPN+3-FL 1 250 1000 1:4 Experiment 14 OPN low + 3-FL high 1 50 1500 1:30 Experiment 15 OPN low + 3-FL medium 1 50 1000 1:20

[0104] 1.8 NO kit to detect the NO content in the supernatant of BV2 cells

[0105] The BV2 cell experimental grouping and nutrient intervention methods were the same as above. After the culture was completed, the cells were placed in a 4°C temperature environment and centrifuged at 1000 rpm for 5 minutes. The supernatant was collected and the NO content in the cell supernatant was determined using the NO kit according to the operating procedures. The principle of the NO kit to detect NO content is that NO is easily oxidized to nitrite ions (NO2 - ), under acidic conditions, NO2 - It can react with sulfonamide to produce diazo compounds, which can then react with naphthylethylenediamine to produce colored compounds. The concentration of the product of this reaction is similar to that of NO2 - There is a linear relationship between the concentration and the absorption peak at 520nm-560nm. - The NO content in the liquid was calculated using the standard curve.

[0106] 1.9 Detection of TNF-α, IL-1β and IL-10 in the supernatant of BV2 cell culture by enzyme-linked immunosorbent assay (ELISA)

[0107] The BV2 cell experimental grouping and nutrient intervention methods were the same as above. After the culture was completed, the cells were placed in a 4°C temperature environment and centrifuged at 1000 rpm for 5 min. The supernatant was collected and the amounts of TNF-α, IL-1β and IL-10 in the cell supernatant were detected according to the operating steps in the kit instructions.

[0108] 1.10 Detection of iNOS and CD206, markers of M1 and M2 phenotypes of BV2 cells

[0109] qRT-PCR was used to detect the mRNA expression levels of iNOS and CD206-related genes and then determine the changes in the content of M1 cell population and M2 cell population. The third-generation BV2 microglial cell suspension was inoculated in a 24-well culture plate with a cell density of 4×10 5Each well was divided into groups according to the experimental design. After treatment, the cells were washed twice with PBS, using 200 μL per well. Digested with trypsin for 2.5 min, and digested with 400 μL of culture medium. The cells were blown up with a pipette and collected in enzyme-free centrifuge tubes. Total RNA of each group of cells was extracted. Reverse transcription was performed using a reverse transcription kit according to the operating procedures to obtain cDNA, and the amplified cDNA was mixed with primers, fluorescent probes and qPCR Master Mix. The corresponding PCR cycle program and parameters were set according to the requirements of qPCR, and the corresponding mRNA expression level was analyzed in the real-time PCR detection system. The specific primer information is as follows:

[0110] iNOS:

[0111] Upstream: 5'-GGCTTGCCCCTGGAAGTTT-3' (SEQ ID NO. 1),

[0112] Downstream: 5′-TGCAAGTGAAATCCGATGTGG-3′ (SEQ ID NO. 2);

[0113] CD206:

[0114] Upstream: 5'-TTCAGCTATTGGACGCGAGG-3' (SEQ ID NO. 3),

[0115] Downstream: 5'-GAATCTGACACCCAGCGGAA-3' (SEQ ID NO. 4);

[0116] GAPDH:

[0117] Upstream: 5'-GGTTGTCTCCTGCGACTTCA-3' (SEQ ID NO. 5),

[0118] Downstream: 5'-TGGTCCAGGGTTTCTTACTCC-3' (SEQ ID NO. 6).

[0119] 1.11 Western blot detection of relative expression of proteins in BV2 cells

[0120] Western blot was used to detect the expression of TLR4, NF-κB, p-NF-κB, IKB-α and p-IKB-α, and the metabolic pathways of TLR4, NF-κB, p-NF-κB, IKB-α and p-IKB-α were studied. The third generation BV2 cell suspension was inoculated in a 6-well culture plate with a cell density of 1×10 5 / well, and treated in groups according to the experimental design. After treatment, wash the cells three times with 4℃ precooled PBS solution, lyse the cells on ice with RIPA lysis buffer containing protease inhibitors, centrifuge at 15000r / min for 15min at 4℃, take the supernatant, use BCA quantitative kit to detect protein concentration, and quantitatively divide the samples. Add 5 times the concentration of SDS-PAGE protein loading buffer and boil in a 100℃ metal water bath for 5min, add 5 times the concentration of SDS-PAGE protein loading buffer and boil in a 100℃ metal water bath for 5min to fully denature the protein. After equal amounts of each group of proteins were loaded, electrophoresis and transfer were performed, and the membrane was blocked with protein-free rapid blocking solution (1×) for 1h. The PVDF membrane was removed and the primary antibody (1:1000) was added and incubated overnight at 4℃. The primary antibody was recovered, washed 3 times with TBST solution, and the corresponding secondary antibody (dilution ratio 1:5000) was added and incubated at room temperature for 1h. Wash again with TBST solution for 3 times, prepare ECL developer (ratio 1:1), expose and develop with chemiluminescence imaging analysis system, and collect images. Use Image J software to analyze the grayscale of protein bands, and the final result is expressed as the grayscale value of target protein / grayscale value of internal reference protein, and the internal reference protein is GAPDH.

[0121] 1.12 Statistical analysis

[0122] All experiments were repeated three times independently, and SPSS22.0 statistical software was used to analyze the experimental data. The experimental results were analyzed by variance homogeneity test, and the statistical differences between the groups were analyzed by one-way ANOVA, and expressed as mean ± standard deviation (x ± SD). P>0.05 indicated no statistical significance, and P<0.05 indicated statistically significant differences.

[0123] 2. Experimental results

[0124] 2.1 Nutrient cytotoxicity assay

[0125] Table 4 shows the effects of different concentrations of nutrients on the cell viability of BV2 cells. The results show that the nutrients have no significant effect on cell viability.

[0126] Table 4 Effects of different nutrients on cell viability of BV2 cells

[0127] Corresponding experiment Dose Group Description OPNμg / mL 3-FLμg / mL OPN:3-FL Cell survival rate% Blank control Blank control group - - 100.77±0.22 Positive Control LPS model group - - 94.36±0.37 Experiment 1 OPN high dose group 500 - 99.90±0.10 Experiment 2 OPN medium dose group 250 - 99.17±0.21 Experiment 3 OPN low dose group 50 - 96.52±0.94 Experiment 4 3-FL high dose group - 1500 96.28±0.31 Experiment 5 3-FL medium dose group - 1000 98.45±0.10 Experiment 6 3-FL medium and low dose group - 250 96.25±0.49 Experiment 7 3-FL low dose group - 50 96.49±0.21 Experiment 8 OPN high + 3-FL high 500 1500 1:3 100.34±0.06 Experiment 9 OPN High + 3-FL Medium 500 1000 1:2 99.00±0.57 Experiment 10 OPN high + 3-FL medium low 500 250 1:0.5 98.07±0.47 Experiment 11 OPN high + 3-FL low 500 50 1:0.1 97.45±0.26 Experiment 12 OPN Medium + 3-FL High 250 1500 1:6 98.90±0.26 Experiment 13 OPN+3-FL 250 1000 1:4 94.11±0.72 Experiment 14 OPN low + 3-FL high 50 1500 1:30 97.93±0.21 Experiment 15 OPN low + 3-FL medium 50 1000 1:20 96.90±1.14

[0128] 2.2 Effects of nutrients on NO production in LPS-stimulated BV2 cells

[0129] In order to measure the effect of mixed nutrition or single nutrition treatment on LPS-induced BV2 cells, the present invention evaluates the secretion of pro-inflammatory chemical signal NO to estimate the effect of regulating microglial polarization. Table 5 shows the effect of different nutritional composition samples containing OPN and 3-FL on the NO production of LPS-stimulated BV2 cells. The results of the NO study showed that compared with the LPS model group, OPN or 3-FL, when used alone, showed the effect of reducing the production of NO. When the two are used in combination, especially when the stoichiometric ratio of OPN:3-FL is controlled from 1:0.1 to 1:30, a significant effect of reducing the NO production induced by LPS can be obtained.

[0130] Specifically, by comparing Experimental Example 2, Experimental Example 5 and Experimental Example 13, it is shown that relative to the LPS model group, cell experiment Examples 2 and 5 reduced the NO content by 0.69 and 1.9, respectively, while cell experiment Example 13 reduced NO by 3.72, which is greater than the sum of the previous two (0.69+1.9), indicating that there is a synergistic effect between the two components, which can synergistically reduce the NO production of BV2 cells stimulated by LPS.

[0131] Table 5 Effects of different nutrient compositions on NO production in BV2 cells stimulated by LPS

[0132]

[0133] 2.3 Effects of nutrients on the production of pro-inflammatory factors in LPS-stimulated BV2 cells

[0134] In order to measure the effects of mixed nutrition or single nutrition treatment on LPS-induced BV2 cells, we evaluated the secretion of proinflammatory factors TNF-α and IL-1β to estimate the effect of regulating microglial polarization. Table 6 shows the effects of different nutritional composition samples containing OPN and 3-FL on the production of proinflammatory factors in LPS-stimulated BV2 cells. The results showed that compared with the LPS model group, when OPN or 3-FL was used alone, the effect of reducing the production of proinflammatory factors was not obvious. When the two were used in combination, the stoichiometric ratio of OPN:3-FL was 1:0.1 to 1:30, a significant effect of reducing the production of LPS-induced proinflammatory factors could be obtained.

[0135] Specifically, by comparing Experimental Example 2, Experimental Example 5 and Experimental Example 13, it is illustrated that relative to the LPS model group, cell experiment Experimental Examples 2 and 5 reduced the TNF-α content by 4.87 and 5.21, and the IL-1β content by 0.93 and 1.07, respectively, while cell experiment Experimental Example 13 reduced TNF-α by 44.45, which is greater than the sum of the first two (4.87+5.21), and reduced IL-1β by 2.98, which is greater than the sum of the first two (0.93+1.07), indicating that there is a synergistic effect between the two components, which can synergistically reduce the production of proinflammatory factors in BV2 cells stimulated by LPS.

[0136] Table 6 Effects of different nutritional compositions on the production of pro-inflammatory factors in BV2 cells stimulated by LPS

[0137]

[0138] 2.4 Effects of nutrients on anti-inflammatory factors in LPS-stimulated BV2 cells

[0139] In order to measure the effects of mixed nutrition or single nutrition treatment on LPS-induced BV2 cells, we evaluated the secretion of the anti-inflammatory factor IL-10 to estimate the effect of regulating microglial polarization. Table 7 shows the effects of different nutritional composition samples containing OPN and 3-FL on the production of anti-inflammatory factors in BV2 cells stimulated by LPS. The results showed that compared with the LPS model group, when OPN or 3-FL was used alone, both showed the effect of increasing the production of anti-inflammatory factors. At the same time, when the two were used in combination, a significant effect of increasing IL-10 production could be obtained.

[0140] Specifically, by comparing Experimental Example 2 (OPN), Experimental Example 5 (3-FL) and Experimental Example 13 (OPN + 3-FL), it was shown that compared with the single nutritional treatment of Experimental Example 2 and Experimental Example 5, the IL-10 content of the mixed nutritional treatment of the cell experiment Experimental Example 13 was significantly increased (P<0.001), indicating that the two can synergistically increase the production of anti-inflammatory factors.

[0141] Table 7 Effects of different nutritional compositions on the production of anti-inflammatory factors in BV2 cells stimulated by LPS

[0142]

[0143] 2.5 Effects of nutrients on iNOS and CD206, markers of M1 and M2 phenotypes of BV2 cells stimulated by LPS

[0144] After activation, microglia eventually polarize into two cell groups with opposite functions, M1 or M2. M1 phenotype microglia mainly release pro-inflammatory cytokines, such as tumor necrosis factor (TNF-α), interleukin 1β (IL-1β) and chemical signal nitric oxide (NO), which may cause inflammatory response in the CNS, leading to neuronal damage and eventually death; M2 phenotype microglia mainly promote damage repair and exert neuroprotective effects by secreting anti-inflammatory cytokines, such as interleukin 10 (IL-10). The main phenotypic marker of M1 microglia is iNOS, and the main phenotypic marker of M2 microglia is CD206.

[0145] In order to measure the effect of mixed nutrition or single nutrition treatment on LPS-induced BV2 cells, we used qRT-PCR to detect the mRNA expression levels of iNOS and CD206-related genes and then measured the changes in the content of M1 cell populations and M2 cell populations to estimate the effect of regulating microglial polarization. Table 8 shows the effects of different nutrient composition samples containing OPN and 3-FL on the mRNA expression levels of BV2 cell markers stimulated by LPS.

[0146] The results showed that compared with the LPS model group, when OPN or 3-FL was used alone, both showed the effect of increasing the relative expression of CD206 and decreasing the relative expression of iNOS. At the same time, when the two were used in combination, the dosage ratio of OPN:3-FL was 1:0.1 to 1:30, and more obvious effects could be obtained.

[0147] Specifically, by comparing Experimental Example 2, Experimental Example 5 and Experimental Example 13, relative to the LPS model group, cell experiment Experimental Examples 2 and 5 reduced the relative expression of iNOS by 2.09 and 2.94, respectively, and increased the relative expression of CD206 by 1.89 and 2.48, while cell experiment Experimental Example 13 reduced the relative expression of iNOS by 6.24, which was greater than the sum of the first two (2.09+2.94), and increased the relative expression of CD206 by 5.89, which was greater than the sum of the first two (1.89+2.48), indicating that there is a synergistic effect between the two components, which can synergistically reduce the polarization of LPS-stimulated BV2 cells to the M1 phenotype.

[0148] Table 8 Effects of different nutritional compositions on the expression levels of BV2 cell marker mRNAs stimulated by LPS

[0149]

[0150] 2.6 Effects of nutrients on the TLR4 / NF-κB signaling pathway in LPS-stimulated BV2 cells

[0151] LPS induces the activation of Toll-like receptors (TLR4), which in essence activates the NF-κB signaling pathway and accelerates the transcription of inflammatory genes. Under normal conditions, NF-κB is in an inactive form and combines with the inhibitor IKB-α in the cytoplasm to form a dimer. After being stimulated by inflammation, the IKB-α protein is phosphorylated and degraded, releasing NF-κB to transfer to the cell nucleus and participate in the expression of several inflammatory factors such as IL-1β and TNF-α. At the same time, these inflammatory factors are involved in regulating the TLR-4 / NF-κB signaling pathway, thus forming a feedback loop and jointly participating in the process of neuroinflammation.

[0152] In order to explore the pathways that affect mixed nutrients or single nutrient treatment on LPS-induced BV2 cells, we extracted the whole protein of each group of cells and used Western blot to detect the expression of TLR4, NF-κB, p-NF-κB, IKB-α and p-IKB-α. Figures 2A to 2F The effects of OPN, 3-FL and their combination on the expression of TLR4 / NF-κB signaling pathway proteins in LPS-stimulated BV2 cells were demonstrated.

[0153] The results showed that compared with the control group, the protein expressions of TLR4, NF-κB p65 and p-NF-κBp65 (phosphorylated NF-κB p65) in the LPS model group were significantly increased (P < 0.0001, Figures 2B to 2D ); Compared with the LPS model group, the expressions of TLR4, NF-κB p65 and p-NF-κB p65 in the high-dose OPN group, high-dose 3-FL group and combination group were inhibited, and the inhibitory effect of the combination was more significant (P < 0.0001, Figures 2B to 2D At the same time, the phosphorylation degree of IKB-α protein was determined. The results showed that compared with the control group, the expression of IKB-α protein in the LPS model group was significantly reduced, and the expression of p-IKB-α (phosphorylated IKB-α) protein was significantly increased, and the difference was statistically significant (P < 0.0001, Figure 2E , 2F ); Compared with the LPS model group, the high-dose OPN group, high-dose 3-FL group and the combination group inhibited the phosphorylation of IKB-α in BV2 cells, and the inhibitory effect of the combination was more significant (P < 0.0001, Figure 2E , 2F ). It can be seen that OPN, 3-FL alone, and especially the combination of OPN and 3-FL can alleviate neuroinflammatory response, which may be achieved through the TLR4 / NF-κB signaling pathway.

Claims

1. Use of a functional nutritional composition in preparing a food that helps regulate brain immunity, characterized in that: The functional nutritional composition comprises the following essential ingredients: 3-fucosyllactose and osteopontin, and in the functional nutritional composition, the mass ratio of 3-fucosyllactose to osteopontin is (0.05-36):

1.

2. The use according to claim 1, characterized in that In the functional nutritional composition, the total weight of the 3-fucosyllactose and osteopontin accounts for more than 60% of the total dry weight of the functional nutritional composition.

3. The use according to claim 1 or 2, characterized in that The contribution to brain immune regulation includes at least one of inhibiting polarization of microglia to M1 phenotype, promoting polarization of microglia to M2 phenotype, and inducing transformation of microglia from M1 phenotype to M2 phenotype.

4. The use according to any one of claims 1 to 3, characterized in that The method of helping brain immune regulation includes at least one of increasing the amount of IL-10 released by microglia, reducing the amount of TNF-α released by microglia, reducing the amount of IL-1β released by microglia, and reducing the amount of NO released by microglia.

5. The use according to any one of claims 1 to 4, characterized in that: The food is an oral preparation, which includes any one or more of tablets, pills, granules, powders, capsules and oral liquids.

6. The use according to any one of claims 1 to 4, characterized in that The food is dairy products, candy, beverages or baked goods.

7. The use according to claim 6, characterized in that The dairy product includes at least one of fermented milk, cheese and milk powder; the candy includes at least one of hard candy, gel candy, crisp candy, compressed candy and inflated candy; the beverage includes at least one of carbonated beverage, tea beverage, coffee beverage, fruit and vegetable juice beverage and lactic acid bacteria beverage; the baked food includes at least one of bread, cake and biscuits.

8. Use of a functional nutritional composition in preparing a food for assisting in improving the health of the central nervous system, characterized in that: The functional nutritional composition comprises the following essential ingredients: 3-fucosyllactose and osteopontin, and in the functional nutritional composition, the mass ratio of 3-fucosyllactose to osteopontin is (0.05-36):

1.

9. The use according to claim 8, characterized in that The auxiliary improvement of central nervous system health includes the improvement of neuroinflammation for non-therapeutic purposes.

10. The use according to claim 8 or 9, characterized in that In the functional nutritional composition, the total weight of the 3-fucosyllactose and osteopontin accounts for more than 60% of the total dry weight of the functional nutritional composition.

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