Nutritional compositions, products comprising same and uses thereof

Abstract

The present application relates to a nutritional composition, a product comprising the same and use thereof in the preparation of a product for alleviating synaptic plasticity impairment of a body. The nutritional composition comprises: docosahexaenoic acid (DHA) and / or a metabolic precursor thereof, sodium hyaluronate (SH), and lactoferrin (LF). When docosahexaenoic acid (DHA), sodium hyaluronate (SH) and lactoferrin (LF) are used in combination, they can synergistically alleviate synaptic plasticity impairment of a body, in particular, increase nerve growth factor (NGF) content and / or brain-derived neurotrophic factor (BDNF) content in PC12 cells of the body, and / or increase expression of synaptic-related proteins in PC12 cells of the body.

Description

Technical Field

[0001] This invention generally relates to the food industry. Specifically, this invention relates to nutritional compositions, products comprising the same, and their use in the preparation of products for mitigating synaptic plasticity damage in the body. Background Technology

[0002] Neurons, as the basic functional units of the brain, consist of dendrites, axons, and cell bodies, working together to receive, transmit, and process information. Neurons connect with each other through synapses at the axon terminals and adjacent dendrites, constructing complex neural networks whose signal transmission relies on the precise regulation of electrochemical pulses. Synaptic plasticity refers to the dynamic adjustment ability of synapses in terms of morphology, structure, and function, including changes in the release of presynaptic neurotransmitters and the regulation of postsynaptic receptor density and potential. This mechanism is the core foundation for the nervous system's adaptation to the environment and its ability to perform learning and memory functions.

[0003] However, synaptic plasticity is easily damaged under the influence of factors such as ischemia, neurotoxins, oxidative stress, or inflammation, manifesting as a reduction in the number of synapses, structural damage, abnormal neurotransmitter release, receptor dysfunction, and abnormal expression of synapse-related proteins (such as PSD-95 and GAP-43). This type of damage is closely related to decreased learning and memory abilities and mood disorders (such as depression), and is also an important link in the development of various neurodegenerative diseases. Studies have shown that brain-derived neurotrophic factor (BDNF) and its signaling pathway play a key role in the regulation of hippocampal synaptic plasticity, suggesting that intervention in the BDNF pathway may help improve memory function.

[0004] In modern society, the fast pace of life and increased stress can lead to a decline in neurological function across different age groups, including headaches and memory loss, which can severely impact daily learning and cognitive activities. Simultaneously, with the aging population, the incidence of neurodegenerative diseases such as Alzheimer's and Parkinson's is rising significantly, with oxidative stress considered a key contributing mechanism. Memory decline is often directly related to hippocampal damage, decreased neurotransmitter levels, and loss of synaptic plasticity. In aging or disease states, synaptic plasticity generally decreases, and the decline in plasticity in brain regions such as the hippocampus, which are associated with short-term memory, can lead to a range of changes from mild cognitive impairment to Alzheimer's disease.

[0005] Current research on stress relief and memory improvement has made some progress, but drug treatments often have side effects, limiting their long-term application. Therefore, developing bioactive nutrients that improve memory function through daily dietary intake has become a research hotspot both domestically and internationally. Especially for the large population requiring long-term care, slowing down synaptic plasticity damage through safe and convenient foods or nutritional combinations is of significant practical importance for maintaining cognitive health and preventing neurological decline.

[0006] Therefore, there is a need to provide nutritional compositions that can effectively slow down synaptic plasticity damage through daily intake. This not only aligns with the development trend of functional foods but also provides a new and feasible approach to supporting brain health and improving quality of life. Summary of the Invention

[0007] This invention was made in view of the aforementioned problems existing in the prior art.

[0008] In a first aspect, the present invention provides a nutritional composition that can effectively slow down synaptic plasticity damage in the body.

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

[0010] In a third aspect, the present invention provides the use of a nutritional composition according to the first aspect of the invention in the preparation of a product for mitigating synaptic plasticity damage in the body.

[0011] Specifically, the present invention is achieved by the following:

[0012] 1. A nutritional composition comprising:

[0013] Docosahexaenoic acid (DHA) and / or its metabolic precursors

[0014] Sodium hyaluronate SH, and

[0015] Lactoferrin LF.

[0016] 2. The nutritional composition as described in Project 1, wherein the total mass of docosahexaenoic acid and its metabolic precursors converted to docosahexaenoic acid is in the ratio of DHA / SH to sodium hyaluronate (1-5):1.

[0017] 3. The nutritional composition as described in any one of items 1-2, wherein the mass ratio of lactoferrin to sodium hyaluronate LF / SH is (2-8):1.

[0018] 4. The nutritional composition as described in any one of items 1-3, wherein the ratio of the total mass of lactoferrin to docosahexaenoic acid (DHA) converted from its metabolic precursors is LF / DHA = (0.4-8):1.

[0019] 5. The nutritional composition as described in any one of items 1-4, wherein the total mass of docosahexaenoic acid and its metabolic precursor converted to docosahexaenoic acid, plus the mass of lactoferrin, is expressed in the mass ratio of sodium hyaluronate (DHA+LF) / SH = (3-13):1.

[0020] 6. The nutritional composition as described in any one of items 1-5, wherein DHA / SH=(1-5):1, LF / SH=(2-8):1, LF / DHA=(0.4-8):1, and (DHA+LF) / SH=(4-13):1.

[0021] 7. The nutritional composition as described in any one of items 1-6, wherein DHA / SH=(2-5):1, LF / SH=(2-8):1, LF / DHA=(1-4):1, and (DHA+LF) / SH=(4-13):1.

[0022] 8. The nutritional composition as described in any one of items 1-7, wherein the weight-average molecular weight of sodium hyaluronate is 2,000-500,000, preferably 5,000-100,000.

[0023] 9. The nutritional composition of any one of items 1-8, wherein the nutritional composition comprises the following:

[0024] Docosahexaenoic acid (DHA) and / or its metabolic precursors

[0025] Sodium hyaluronate SH, and

[0026] Lactoferrin LF.

[0027] 10. A product comprising a nutritional composition as described in any one of items 1-9.

[0028] 11. The product described in item 10 is a food product, such as a general food product, a functional food product, a health food product, and / or a health supplement product.

[0029] 12. The use as described in item 11, wherein the food is a powdered reconstituteable food, a baked food, a beverage, milk, dairy products, or pasta products.

[0030] 13. The use as described in Item 11, wherein the food is infant formula (e.g., baby formula, follow-up formula, toddler formula), children's formula, maternal and infant food, adult food, and / or food for the middle-aged and elderly.

[0031] 14. Use of the nutritional composition described in any one of items 1-9 in the preparation of products for mitigating synaptic plasticity damage in the body.

[0032] 15. The use as described in item 14, wherein mitigating synaptic plasticity damage in the body includes increasing the levels of nerve growth factor (NGF) and / or brain-derived neurotrophic factor (BDNF) in PC12 cells of the body.

[0033] 16. The use as described in any one of items 14-15, wherein mitigating synaptic plasticity damage in the body comprises increasing the expression of synaptic-associated proteins in PC12 cells of the body, such as synaptophysin SYN and / or growth-associated protein 43, i.e., GAP-43.

[0034] 17. The use as described in any one of items 14-16, wherein when taken orally in humans, docosahexaenoic acid is used at a concentration of 0.01-2.5 mg / ml, for example 0.02-1 mg / ml, sodium hyaluronate is used at a concentration of 0.01-2 mg / ml, for example 0.02-1 mg / ml, and lactoferrin is used at a concentration of 0.005-1 mg / ml, for example 0.01-0.5 mg / ml.

[0035] 18. The use as described in any one of items 14-17, wherein the target of the reduction of synaptic plasticity damage includes infants, older infants, toddlers, infants, children, adolescents, adults, young adults, teenagers, middle-aged people, and / or the elderly.

[0036] The inventors discovered that when docosahexaenoic acid (DHA) and / or its metabolic precursors, sodium hyaluronate (SH) and lactoferrin (LF) are used in combination, they can synergistically slow down synaptic plasticity damage in the body, particularly by increasing the levels of nerve growth factor (NGF) and / or brain-derived neurotrophic factor (BDNF) in PC12 cells (adrenal pheochromocytoma), and / or increasing the expression of synapse-related proteins such as synaptophysin SYN and / or growth-associated protein 43 (GAP-43) in PC12 cells. Detailed Implementation

[0037] Unless otherwise specified, the technical terms in this specification have the same meaning as those generally understood by those skilled in the art; however, in case of any conflict, the definitions in this specification shall prevail.

[0038] As used herein, the following terms have the following meanings.

[0039] The term "infant" refers to a person aged 0 to 6 months.

[0040] The term "older baby" refers to people aged 6 to 12 months.

[0041] The term "infant" refers to people aged 12 to 36 months.

[0042] The term "infant" refers to people aged 0-36 months.

[0043] The term "child" refers to people aged 3 to 6.

[0044] The term "teenager" refers to people aged 7-17.

[0045] The term "adult" refers to a person who is 18 years of age or older.

[0046] The term "young people" refers to people aged 18-40.

[0047] The term "teenager" refers to people aged 7 to 40.

[0048] The term "middle-aged" refers to people aged 41-65.

[0049] The term “elderly” or “senior citizen” refers to people who are 65 years of age or older.

[0050] The term "infant formula" as used in this article encompasses infant formula, follow-up formula, and toddler formula. Generally, infant formula is used as a breast milk substitute from birth, follow-up formula is used from 6-12 months after birth, and toddler formula is used from 12-36 months after birth.

[0051] The term "infant formula" refers to liquid or powdered products made primarily from milk and milk protein products or soy and soy protein products, with the addition of appropriate amounts of vitamins, minerals, and / or other ingredients, produced and processed solely using physical methods. It is suitable for consumption by healthy infants, and its energy and nutrient content can meet the normal nutritional needs of infants aged 0-6 months.

[0052] The term "follow-up formula" refers to liquid or powdered products made primarily from milk and milk protein products or soy and soy protein products, with the addition of appropriate amounts of vitamins, minerals, and / or other ingredients, produced and processed solely using physical methods. These products are suitable for older infants, and their energy and nutrient content can meet some of the nutritional needs of normal older infants aged 6-12 months.

[0053] The term "infant formula" refers to liquid or powdered products made primarily from milk and milk protein products or soybeans and soy protein products, with the addition of appropriate amounts of vitamins, minerals, and / or other ingredients, produced and processed using only physical methods. These products are suitable for infants and their energy and nutrient content can meet some of the nutritional needs of normal infants aged 12-36 months.

[0054] The term "breast milk" should be understood as the mother's breast milk or colostrum.

[0055] The term "infant or toddler exclusively breastfed" has the common meaning of referring to an infant whose nutrients and / or energy are derived primarily from human breast milk.

[0056] The term "infants / followers / toddlers primarily fed with infant formula" has a general meaning, referring to infants or toddlers whose nutrients and / or energy are primarily derived from infant formula, follow-up milk, or growth milk produced by physical methods. The term "primarily" means at least 50%, for example, at least 75%, of those nutrients and / or energy.

[0057] In this application, “non-therapeutic use” and “non-medical therapeutic use” are synonymous and can be used interchangeably, referring to use for nutrition and / or health care through daily consumption.

[0058] Furthermore, in the context of this invention, the terms "comprising" or "including" do not exclude other possible elements. The compositions of this invention (including the various embodiments described herein) may comprise, consist of, or consist substantially of the following elements: the essential elements and necessary limitations of the invention as described herein, and any other or optional ingredients, components, or limitations as described herein or as otherwise desired.

[0059] The individuals described in this invention are applicable to normal humans and may be infants and / or older infants, and / or toddlers, and / or children, and / or young adults, and / or middle-aged adults, and / or elderly people.

[0060] Unless otherwise specified, in this application, when referring to the amount or ratio of a substance or component, it is in terms of mass.

[0061] It should be noted that the various aspects, features, implementation methods, experimental examples, and advantages described in this application are compatible and / or can be combined together.

[0062] This invention relates to nutritional compositions, products comprising the same, and their uses.

[0063] The present invention will now be described in detail.

[0064] Nutritional composition

[0065] In a first aspect, the present invention relates to a nutritional composition comprising:

[0066] Docosahexaenoic acid (DHA) and / or its metabolic precursors

[0067] Sodium hyaluronate SH, and

[0068] Lactoferrin LF.

[0069] Docosahexaenoic acid (DHA) refers to cis-4,7,10,13,16,19-docosahexaenoic acid, a straight-chain fatty acid containing 22 carbon atoms and 6 carbon-carbon double bonds, with the molecular formula C2. 22 H 32 O2. In this article, the terms "docosahexaenoic acid" and "DHA" are used interchangeably.

[0070] In this application, "metabolic precursor" has a well-known meaning in the art; when referring to a substance as a "metabolic precursor," it means a compound that can be converted into that substance in the biochemical reactions of an organism, such as the human body. It is the "starting point" or "intermediate raw material" in a metabolic pathway. Therefore, a metabolic precursor of DHA refers to a compound that can be converted into DHA in the biochemical reactions of an organism, such as the human body.

[0071] DHA can be used in its free acid form and / or its metabolic precursor form. There are no particular restrictions on the metabolic precursors of DHA, and it can be any human-acceptable / edible ester approved by regulations, such as one or more of the following forms: triglycerides (e.g., fish oil triglycerides, concentrated triglycerides (rTG), or algal oil triglycerides), ethyl esters, phospholipids, monoacylglycerides, etc.

[0072] In this application, for convenience, "docosahexaenoic acid and / or its metabolic precursors" are sometimes referred to as "docosahexaenoic acid" or "DHA".

[0073] Sodium hyaluronate (SH) is a high-molecular-weight linear mucopolysaccharide produced by the polymerization of glucuronic acid and acetylaminohexose into a disaccharide. Its chemical formula is (C...). 14 H 20 NO 11 Na) n .

[0074] Lactoferrin (LF) is a multifunctional natural iron-binding glycoprotein that is widely found in mammalian whey and various biological fluids. Its molecular weight is approximately 70-80 kDa and it is composed of about 700 amino acids.

[0075] The inventors discovered that when docosahexaenoic acid (DHA) and / or its metabolic precursors, sodium hyaluronate (SH), and lactoferrin (LF) are used in combination, they can synergistically slow down synaptic plasticity damage in the body, particularly by increasing the levels of nerve growth factor (NGF) and / or brain-derived neurotrophic factor (BDNF) in PC12 cells (pheochromocytoma), and / or increasing the expression of synapse-related proteins such as synaptophysin SYN and / or growth-associated protein 43 (GAP-43) in PC12 cells. Furthermore, the combination of docosahexaenoic acid (DHA), sodium hyaluronate (SH), and lactoferrin (LF) can also enhance the viability of PC12 cells.

[0076] In this embodiment, the weight-average molecular weight of sodium hyaluronate is not particularly limited and may be, for example, 2,000-500,000, preferably 5,000-100,000, such as 2,000, 5,000, 10,000, 20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000, 100,000, 150,000, 200,000, 250,000, 300,000, 350,000, 400,000, 450,000, 500,000, or within the range defined by any two thereof. The weight-average molecular weight can be measured by gel permeation chromatography.

[0077] In some embodiments, lactoferrin may be provided as a lactoferrin (LF)-rich nutrient, such as lactoferrin-rich whey protein concentrate and / or lactoferrin powder, preferably lactoferrin powder.

[0078] In some embodiments, the ratio of the total mass of docosahexaenoic acid (DHA) and its metabolic precursors converted to docosahexaenoic acid to the mass of sodium hyaluronate is (1-5):1; for example, the total mass of docosahexaenoic acid and its metabolic precursors converted to docosahexaenoic acid relative to 1 part by mass of sodium hyaluronate may be 1, 2, 3, 4, or 5 parts by mass, or within the range defined by any two of them.

[0079] In some embodiments, the mass ratio of lactoferrin to sodium hyaluronate is (2-8):1; for example, the amount of lactoferrin may be 2, 3, 4, 5, 6, 7, or 8 parts by mass relative to 1 part by mass of sodium hyaluronate, or within the range defined by any two of these.

[0080] In some embodiments, the mass ratio of lactoferrin to the total mass of docosahexaenoic acid (DHA) converted from its metabolic precursors is LF / DHA = (0.4-8):1; for example, relative to 1 part by mass of the total mass of DHA converted from its metabolic precursors, lactoferrin may be 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0 parts by mass, or within the range defined by any two thereof.

[0081] In some embodiments, the mass ratio of the sum of the total mass of docosahexaenoic acid and its metabolic precursors converted to docosahexaenoic acid and the mass of lactoferrin to sodium hyaluronate (DHA+LF) / SH = (3-13):1; for example, relative to 1 part by mass of sodium hyaluronate, the sum of the total mass of docosahexaenoic acid and its metabolic precursors converted to docosahexaenoic acid and the mass of lactoferrin may be 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 parts by mass, or within the range defined by any two of them.

[0082] In some preferred embodiments, DHA / SH = (1-2):1, LF / SH = (2-8):1, LF / DHA = (1-8):1, and (DHA+LF) / SH = (3-9):1; within these ranges, the levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in PC12 cells can be synergistically increased, as well as the expression of synapse-related proteins such as synaptophysin SYN and growth-associated protein 43 (GAP-43) in PC12 cells can be synergistically increased.

[0083] In some preferred embodiments, DHA / SH = (1-5):1, LF / SH = (2-8):1, LF / DHA = (0.4-8):1, and (DHA+LF) / SH = (4-13):1; within these ranges, the synergistic effect in mitigating synaptic plasticity damage is more significant, and the levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in PC12 cells are more significantly increased, as are the expression of synaptic-related proteins such as synaptophysin SYN and growth-associated protein 43 (GAP-43) in PC12 cells.

[0084] In some further preferred embodiments, DHA / SH = (2-5):1, LF / SH = (2-8):1, LF / DHA = (1-4):1, and (DHA+LF) / SH = (4-13):1; within these ranges, the synergistic effect in mitigating synaptic plasticity damage is more significant, and the levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in PC12 cells are more significantly increased, as are the expression of synaptic-related proteins such as synaptophysin SYN and growth-associated protein 43 (GAP-43) in PC12 cells.

[0085] In some embodiments, the nutritional composition may consist of the following:

[0086] Docosahexaenoic acid (DHA) and / or its metabolic precursors

[0087] Sodium hyaluronate SH, and

[0088] Lactoferrin LF.

[0089] The nutritional compositions of the present invention can be prepared by any suitable method known in the art. For example, the nutritional compositions can be prepared by mixing the components together. The mixing can be carried out by any suitable equipment and methods known in the art.

[0090] product

[0091] In a second aspect, the present invention relates to products comprising a nutritional composition according to the first aspect of the present invention.

[0092] All descriptions above regarding the nutritional composition of the first aspect of the present invention are applicable here and will not be repeated here.

[0093] The product may be in the form of a solid (e.g., powder), liquid, or semi-solid (i.e., a solid-liquid mixture).

[0094] The product may be food or medicine.

[0095] In some embodiments, the product is a food product, such as a regular food product, a functional food product, a health food product, and / or a health supplement.

[0096] In some embodiments, the food may be a powdered reconstituted food, baked goods, beverages, milk, dairy products, or pasta products.

[0097] In some embodiments, the food product may be milk powder.

[0098] In some embodiments, the food may be infant formula (e.g., baby formula, follow-up formula, toddler formula, such as infant formula milk powder), children's formula, maternal and infant food (e.g., formula milk powder for pregnant women), adult food, and / or food for the middle-aged and elderly.

[0099] In some embodiments, the product is a pharmaceutical product. The pharmaceutical product may be any dosage form, such as a solid dosage form (powder, tablet, etc.) or a liquid dosage form. In some embodiments, depending on the dosage form, the pharmaceutical product may also contain a pharmaceutically acceptable carrier and / or excipients.

[0100] There is no particular limitation on the amount of the nutritional composition added to the product (e.g., food), and it can be appropriately selected according to the intended purpose (e.g., the target user group). For example, in some embodiments, the nutritional composition may account for 0.1-100% by mass of the product, such as 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100% by mass, or a range defined by any two thereof. In some embodiments, the nutritional composition may account for, for example, 0.1-30% by mass of the product; when the content of the nutritional composition in the food is within the above-mentioned range, it can effectively slow down synaptic plasticity damage in the body while also balancing other nutrients required by the human body.

[0101] In addition to the components described above for the nutritional composition, the product (e.g., food) may also contain other ingredients such as proteins, carbohydrates, fats, vitamins, minerals, etc., which are often found in formulated foods such as infant formula, such as milk powder. Examples include milk fat globule membrane protein, cow and / or goat milk (e.g., raw cow and / or goat milk), whole milk powder, skim milk powder, demineralized whey powder, demineralized whey liquid, whey protein powder, whey protein concentrate, milk fat globule membrane protein powder, osteopontin (OPN), α-lactalbumin, hydrolyzed whey protein, soy protein, and lactose. Soybean oil, coconut oil, flaxseed oil, walnut oil, sunflower seed oil, 1,3-dioleoyl-2-palmitoylglycerol triglyceride, linoleic acid, alpha-linolenic acid, d-alpha-tocopherol and / or d-alpha-tocopherol derivatives, vitamin A, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, biotin, folic acid, inositol, niacin, pantothenic acid, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, taurine, L-carnitine, lutein, nucleotides, 1,3-dioleoyl-2-palmitoylglycerol triglyceride, etc.

[0102] use

[0103] In a third aspect, the present invention relates to the use of a nutritional composition according to the first aspect of the invention in the preparation of a product for mitigating synaptic plasticity damage in the body, or for non-therapeutic purposes (nutritional and / or health care use) for mitigating synaptic plasticity damage in the body.

[0104] All descriptions above relating to the first and second aspects of the present invention are applicable here and will not be repeated here.

[0105] In some embodiments, mitigating synaptic plasticity damage in the body includes increasing the viability of PC12 cells, such as cell survival rate.

[0106] In some embodiments, the mitigation of synaptic plasticity damage includes increasing the levels of nerve growth factor (NGF) and / or brain-derived neurotrophic factor (BDNF) in PC12 cells, for example, increasing the levels of NGF and BDNF in PC12 cells.

[0107] In some embodiments, mitigating synaptic plasticity damage includes increasing the expression of synapse-related proteins in PC12 cells. In some embodiments, the synapse-related proteins are, for example, synaptophysin SYN and / or growth-associated protein 43, i.e., GAP-43.

[0108] In some implementations, when taken orally by the human body,

[0109] Dodecenoic acid can be used, for example, at concentrations of 0.01-2.5 mg / ml, such as 0.02-1 mg / ml, such as 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, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5 mg / ml, or a range defined by any two thereof;

[0110] Sodium hyaluronate can be used, for example, at concentrations of 0.01-2 mg / ml, such as 0.02-1 mg / ml, such as 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, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 mg / ml, or a range defined by any two thereof;

[0111] Lactoferrin can be used, for example, at concentrations of 0.005-1 mg / ml, such as 0.01-0.5 mg / ml, such as 0.005, 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 mg / ml, or a range defined by any two of these concentrations.

[0112] In some implementations, the target population for mitigating synaptic plasticity damage includes, but is not limited to, the following groups: infants, older infants, toddlers, infants, children, adolescents, adults, young adults, teenagers, middle-aged adults, and / or the elderly.

[0113] Experimental Example

[0114] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to experimental examples. It should be understood that the specific experimental examples described herein are merely illustrative and not intended to limit the invention. Unless otherwise specified, the reagents, methods, and equipment used in this invention are conventional in the art.

[0115] 1. Experiment

[0116] 1.1 Experimental materials and reagents

[0117] Docosahexaenoic acid (DHA): Sigma-Aldrich, DHA content ≥98% by weight

[0118] Sodium Hyaluronate (SH): Bloomage Biotechnology Co., Ltd., Molecular Weight (Mw): 106 kDa, Sodium Hyaluronate Content 96.3% by Mass

[0119] Lactoferrin (LF): Hilmar, Hilmar 1000 lactoferrin, lactoferrin content 97.2% by weight

[0120] PC12 cells: Cell Bank of Chinese Academy of Sciences

[0121] DMEM medium: Gibco, USA

[0122] Fetal bovine serum: Gibco, USA

[0123] Heat-inactivated horse serum: Gibco, USA

[0124] Penicillin-Streptomycin: Beijing Femob Biotechnology Co., Ltd.

[0125] Cell Counting Kit-8 (CCK-8): Beyotime Biotechnology Co., Ltd., Product Code C0038

[0126] NGF Detection Kit: Shanghai Jianglai Biotechnology Co., Ltd., JL12881

[0127] BDNF Detection Kit: Shanghai Jianglai Biotechnology Co., Ltd., Brand No. JL12910

[0128] SYN Detection Kit: Shanghai Jianglai Biotechnology Co., Ltd., Brand No. JL25161

[0129] GAP-43 Detection Kit: Shanghai Jianglai Biotechnology Co., Ltd., Product Code JL13777

[0130] 1.2 Experimental Methods

[0131] PC12 cells are tumor cells of the sympathetic nervous system, possessing general characteristics of neuroendocrine cells. They exhibit a neuronal phenotypic response to nerve growth factor (NGF), accompanied by physiological and biochemical changes. They share many structural and functional similarities with dopaminergic neurons and are characterized by rapid growth and proliferation, short culture cycles, and easily controllable culture conditions. Currently, they are frequently used internationally to study the physiology, pathology, and pharmacology of neurons. The neuronal oxidative damage model established based on PC12 cells is an important pharmacological model, often used to screen for active ingredients in vitro. Oxidative stress is one of the common mechanisms leading to neuronal damage. Treatment of PC12 cells with oxidants (such as H2O2) can induce synaptic damage, manifested as decreased expression of synaptic-related proteins, altered cell morphology, and neurotransmitter imbalance.

[0132] Therefore, this study used H2O2 as an inducer of oxidative damage to construct a PC12 cell synaptic injury model. The experimental group was supplemented with DHA, SH, and LF to observe whether they had a protective effect in mitigating synaptic plasticity damage in PC12 cells.

[0133] 1.3 Cell Culture and Grouping:

[0134] PC12 cell resuscitation and culture: PC12 cells were removed from the liquid nitrogen container and quickly placed in a 37°C water bath to thaw completely within 1-1.5 minutes. The thawed cryopreservation solution was pipetted into centrifuge tubes, and complete culture medium was added to a final volume of 10 mL (complete culture medium was prepared from DMEM medium, fetal bovine serum, heat-inactivated horse serum, and penicillin-streptomycin in a volume ratio of 84:5:10:1). After thorough mixing, the cells were centrifuged at 1000 rpm for 5 minutes. The supernatant was carefully removed, and 3 mL of complete culture medium was added to resuspend the cells. The volume was then increased to 10 mL of complete culture medium, and the cells were incubated at 37°C in a 5% CO2 incubator. Cells were divided into 14 groups as shown in Table 1 and treated differently: ① blank control group; ② positive control group (hydrogen peroxide stimulation group), in which hydrogen peroxide was added to the culture medium to a final concentration of 100 μmol / L and treated for 6 h; ③ experimental group (divided into comparative and example groups), in which 100 μL of lactoferrin, DHA, SH and a combination of two or three were added to each well of each group, and the specific concentrations are shown in Table 1. After each group was treated for 24 h, hydrogen peroxide was added to a final concentration of 100 μmol / L and treated for 6 h.

[0135] Table 1 Experimental Groups

[0136]

[0137] 1.4 Detection of the effect of nutrients on cell viability:

[0138] To investigate the effects of lactoferrin, DHA, SH, and their combined use on PC12 cell viability, the cell counting kit-8 (CCK-8) method was used to determine PC12 cell viability. Following the instructions, PC12 cells were seeded into 96-well plates at 1 × 10⁶ cells per well. 4 Cells were grouped and treated in 96-well plates. 10 μl of CCK-8 kit solution was added to each well, and the plates were incubated at 37°C for 1 h. The absorbance at 450 nm was measured using a BioTek SynergyHTX microplate reader, and cell viability was calculated using the following formula:

[0139] Cell viability (%) = [Absorbance value of positive control group or experimental group - Absorbance value of control group (containing culture medium only, no cells)] / [Absorbance value of blank control group - Absorbance value of control group (containing culture medium only, no cells)] × 100%

[0140] 1.5 Measurement of neurotrophic factor-related indicators:

[0141] Neurotrophic factors are a class of proteins that play important roles in the survival, development, and functional maintenance of neurons. Among them, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are the two most important neurotrophic factors, playing key roles in neuronal survival, differentiation, synaptic plasticity, and functional regulation.

[0142] NGF primarily supports the development, survival, and maintenance of neurons, providing a foundation for synapse formation and plasticity. After nerve injury, it can also promote the regeneration and repair of damaged neurons, helping to restore nerve function and synaptic connections.

[0143] BDNF is a key factor involved in most forms of synaptic plasticity. It promotes neurotransmitter release and enhances receptor function, thereby improving synaptic transmission efficiency. BDNF is also a key molecule inducing long-term potentiation (LTP)—a form of synaptic plasticity closely related to learning and memory—primarily by activating downstream signaling pathways through its receptor TrkB, promoting the formation and maintenance of LTP.

[0144] In synaptic plasticity, NGF and BDNF have complementary and synergistic effects: NGF provides basic support for synaptic plasticity, while BDNF directly regulates synaptic transmission and drives the plasticity process. Together, they maintain the normal function of the nervous system and play an important role in physiological processes such as learning, memory, and nerve damage repair.

[0145] Therefore, the effects of nutrients on the levels of NGF and BDNF in PC12 cells were specifically studied in the experiment.

[0146] Specifically, after processing as described in "1.3 Cell Culture and Grouping" above, cells were collected in centrifuge tubes and stored at -80℃ for later use. The NGF and BDNF levels in PC12 cells were measured according to the kit instructions.

[0147] 1.6 Determination of synaptic plasticity-related protein content:

[0148] Synaptic plasticity-related proteins play a crucial role in the function and adaptation of the nervous system, influencing processes such as nerve signal transmission, learning, and memory by regulating synaptic structure and function. Among them, synaptophysin (SYN) and growth-associated protein 43 (GAP-43) are two important synaptic-related proteins.

[0149] SYN is a phosphoprotein widely distributed on the presynaptic vesicle membrane of nerve terminals, primarily involved in synaptic vesicle docking and fusion, as well as Ca²⁺-dependent neurotransmitter release and vesicle circulation. Its expression level reflects synaptic activity and quantity, serving as an important marker of synaptic function and plasticity. Furthermore, SYN also participates in axonal growth, playing a role in synapse formation and maintenance.

[0150] GAP-43 is a protein closely related to axonal growth and nerve ending plasticity. It is highly expressed during neuronal development and is mainly found in axons and presynaptic membranes. It supports neurite extension and regeneration by promoting pseudopodia growth and actin accumulation on the cell membrane during axonal growth, branching, and guidance; its upregulation can further promote axonal budding and synapse formation. Therefore, GAP-43 is a key factor in synaptic plasticity, neural development, and damage repair, and is closely related to learning and memory functions in regions such as the hippocampus and association cortex of the adult brain.

[0151] Therefore, the effects of nutrients on the levels of SYN and GAP-43 in PC12 cells were specifically studied in the experiment.

[0152] Specifically, after processing as described in "1.3 Cell Culture and Grouping" above, cells were collected in centrifuge tubes and stored at -80℃ for later use. Following the kit instructions, the levels of SYN and GAP-43 in PC12 cells were measured.

[0153] 1.7 Statistical Analysis:

[0154] Statistical analysis was performed on the experimental data. One-way ANOVA (Duncan's test) was used to analyze statistical differences between groups, and the results were expressed as mean ± standard deviation (x ± SD). P > 0.05 indicated no significant difference, and P < 0.05 indicated a significant difference.

[0155] 2. Experimental Results

[0156] 2.1 Effects of nutrients on nerve cell survival:

[0157] Table 2 shows the effect of nutrients on nerve cell survival.

[0158] Table 2 Cell viability

[0159]

[0160] As shown in Table 2, the survival rate of PC12 cells treated with H2O2 in the positive control group was approximately 50%. Compared with the positive control group, all nutrient treatment groups significantly improved the survival rate of H2O2-induced PC12 cells.

[0161] 2.2 Effects of nutrients on neurotrophic factor-related indicators:

[0162] Table 3 shows the effects of nutrients on the secretion of neurotrophic factors NGF and BDNF by PC12 cells.

[0163] Table 3 Effect of the composition on the content of neurotrophic factors

[0164]

[0165] Note: Different letters indicate significant differences between treatments (P<0.05).

[0166] As shown in Table 3, compared with the blank control group, the positive control group treated with H2O2 significantly reduced the intracellular NGF and BDNF levels (P < 0.05). Compared with the positive control group, the NGF and BDNF levels in the groups treated with DHA, SH, or LF alone or in combination were significantly increased (P < 0.05). Compared with Comparative Example 1, the neurotrophic factor levels in Comparative Example 2 were increased (P < 0.05), suggesting that adding SH to DHA can further increase NGF and BDNF. Comparative Examples 3 and 4 added LF to Comparative Example 1, and the results showed that the NGF and BDNF levels in Comparative Examples 3 and 4 were significantly increased compared with Comparative Example 1 (P < 0.05), suggesting that the combination of DHA and LF has a significant effect on promoting the secretion of neurotrophic factors. Comparative Examples 5 and 6 further increased the DHA content to Comparative Example 2, and the results showed that the NGF content was further increased, but excessively high DHA content did not further increase BDNF.

[0167] Based on the comparative examples, DHA, SH, and LF were added simultaneously in Experiments 1 to 6, and the results showed that the composition could further increase the content of NGF and BDNF (P < 0.05). Furthermore, there was a synergistic effect among the components DHA, SH, and LF in Examples 1-6.

[0168] Specifically, taking Example 1 as an example, it is equivalent to subtracting Comparative Example 1 from the combination of Comparative Examples 2 and 3. Compared with the positive control group, Comparative Examples 1, 2, and 3 increased NGF by 12.22, 20.84, and 21.01, respectively, while Example 1 increased NGF content by 53.46, which is greater than the value obtained by Comparative Example 2 + Comparative Example 3 - Comparative Example 1 (29.63); Comparative Examples 1, 2, and 3 increased BDNF by 7.63, 13.69, and 15.97, respectively, while Example 1 increased BDNF content by 40.36, which is greater than the value obtained by Comparative Example 2 + Comparative Example 3 - Comparative Example 1 (22.03); the results indicate that there is a synergistic effect between DHA, SH, and LF, which jointly promote the secretion of NGF and BDNF in cells, promote synaptic plasticity, and play a role in mitigating synaptic plasticity damage.

[0169] Similarly, the same conclusions can be drawn for Examples 2-6.

[0170] 2.3 Effects of nutrients on synaptic plasticity-related proteins:

[0171] Table 4 shows the effects of nutrients on the expression of synaptic plasticity-related proteins SYN and GAP-43 in PC12 cells.

[0172] Table 4. Effect of the composition on the content of synaptic plasticity-related proteins

[0173]

[0174] Note: Different letters indicate significant differences between treatments (P<0.05).

[0175] As shown in Table 4, compared with the blank control group, the expression of SYN and GAP-43 in the H2O2-treated positive control group was significantly decreased (P < 0.05). Compared with the positive control group, the expression of SYN and GAP-43 in the DHA, SH, and LF treatment groups, alone or in combination, was significantly increased (P < 0.05). Compared with Comparative Example 1, the content of synaptic plasticity-related proteins in Comparative Example 2 was increased (P < 0.05), suggesting that adding SH to DHA can further enhance the expression of SYN and GAP-43. Comparative Examples 3 and 4 added LF to Comparative Example 1, and the results showed that the expression of SYN and GAP-43 in Comparative Examples 3 and 4 was significantly increased compared with Comparative Example 1 (P < 0.05), suggesting that the combination of DHA and LF has a significant effect on promoting the expression of synaptic plasticity-related proteins. Comparative Examples 5 and 6 further increased the content of DHA to Comparative Example 2, and the results showed that the expression of SYN and GAP-43 was further increased (P < 0.05).

[0176] Based on the comparative examples, DHA, SH, and LF were added simultaneously in Experiments 1 to 6. The results showed that the composition could further enhance the expression of SYN and GAP-43 (P < 0.05). Furthermore, there was a synergistic effect between the components DHA, SH, and LF in Examples 1-6 in terms of NGF and BDNF.

[0177] Specifically, taking Example 1 as an example, it is equivalent to subtracting Comparative Example 1 from the combination of Comparative Examples 2 and 3. Compared with the positive control group, Comparative Examples 1, 2, and 3 increased SYN by 2.14, 3.3, and 4.34, respectively, while Example 1 increased SYN content by 11.27, which is greater than the value obtained by Comparative Example 2 + Comparative Example 3 - Comparative Example 1 (5.5); Comparative Examples 1, 2, and 3 increased GAP-43 by 4.08, 8.04, and 8.55, respectively, while Example 1 increased GAP-43 content by 26.37, which is greater than the value obtained by Comparative Example 2 + Comparative Example 3 - Comparative Example 1 (12.51); the results indicate that there is a synergistic effect between DHA, SH, and LF, and their combined use is more effective in upregulating the expression of SYN and GAP-43 than treatment with any one or two components alone, and has a stronger function in maintaining synaptic plasticity.

[0178] Similarly, the same conclusions can be drawn for Examples 2-6.

[0179] The above description is merely an exemplary embodiment of the present invention. It should be noted that those skilled in the art can make improvements to the present invention without departing from the inventive concept, and all such improvements fall within the scope of protection of the present invention.

Claims

1. A nutritional composition comprising: Docosahexaenoic acid (DHA) and / or its metabolic precursors Sodium hyaluronate SH, and Lactoferrin LF.

2. The nutritional composition of claim 1, wherein the total mass of docosahexaenoic acid and its metabolic precursors converted to docosahexaenoic acid is in the mass ratio of sodium hyaluronate (DHA / SH) of (1-5):

1.

3. The nutritional composition according to any one of claims 1-2, wherein the mass ratio of lactoferrin to sodium hyaluronate LF / SH is (2-8):

1.

4. The nutritional composition according to any one of claims 1-3, wherein the mass ratio of lactoferrin to the total mass of docosahexaenoic acid (DHA) converted from its metabolic precursors is LF / DHA = (0.4-8):

1.

5. The nutritional composition according to any one of claims 1-4, wherein the total mass of docosahexaenoic acid and its metabolic precursor converted to docosahexaenoic acid, plus the mass of lactoferrin, is expressed in the mass ratio of sodium hyaluronate (DHA+LF) / SH = (3-13):

1.

6. The nutritional composition according to any one of claims 1-5, wherein DHA / SH=(1-5):1, LF / SH=(2-8):1, LF / DHA=(0.4-8):1, and (DHA+LF) / SH=(4-13):

1.

7. The nutritional composition according to any one of claims 1-6, wherein DHA / SH=(2-5):1, LF / SH=(2-8):1, LF / DHA=(1-4):1, and (DHA+LF) / SH=(4-13):

1.

8. The nutritional composition according to any one of claims 1-7, wherein the weight-average molecular weight of sodium hyaluronate is 2,000-500,000, preferably 5,000-100,000.

9. The nutritional composition according to any one of claims 1-8, wherein the nutritional composition comprises: Docosahexaenoic acid (DHA) and / or its metabolic precursors Sodium hyaluronate SH, and Lactoferrin LF.

10. A product comprising the nutritional composition as described in any one of claims 1-9.

11. The product as described in claim 10 is a food product, such as a regular food product, a functional food product, a health food product, and / or a health supplement product.

12. The use as described in claim 11, wherein the food is a powdered reconstituteable food, a baked food, a beverage, milk, dairy products, or pasta products.

13. The use as described in claim 11, wherein the food is infant formula (e.g., baby formula, follow-up formula, toddler formula), children's formula, maternal and infant food, adult food, and / or food for the middle-aged and elderly.

14. Use of the nutritional composition according to any one of claims 1-9 in the preparation of products for mitigating synaptic plasticity damage in the body.

15. The use as described in claim 14, wherein mitigating synaptic plasticity damage in the body comprises increasing the levels of nerve growth factor (NGF) and / or brain-derived neurotrophic factor (BDNF) in PC12 cells of the body.

16. The use as described in any one of claims 14-15, wherein mitigating synaptic plasticity damage in the body comprises increasing the expression of synaptic-associated proteins in PC12 cells of the body, such as synaptophysin SYN and / or growth-associated protein 43, i.e., GAP-43.

17. The use as described in any one of claims 14-16, wherein when taken orally in humans, docosahexaenoic acid is used at a concentration of 0.01-2.5 mg / ml, for example 0.02-1 mg / ml, sodium hyaluronate is used at a concentration of 0.01-2 mg / ml, for example 0.02-1 mg / ml, and lactoferrin is used at a concentration of 0.005-1 mg / ml, for example 0.01-0.5 mg / ml.

18. The use as described in any one of claims 14-17, wherein the target of the reduction of synaptic plasticity damage includes infants, older infants, toddlers, young children, children, adolescents, adults, young adults, teenagers, middle-aged people, and / or the elderly.

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