Use of nutritional composition to assist in improving memory ability

Abstract

The present invention belongs to the field of food, and specifically relates to the use of a nutritional composition to assist in improving memory ability. The present invention provides a use of a nutritional composition in preparing a food that is ingested by a mother to assist in improving the memory ability of an offspring, the nutritional composition comprising the following essential components: active folic acid substances and N-acetylneuraminic acid; and, in the nutritional composition, the mass ratio of the active folic acid substances to the N-acetylneuraminic acid is (0.0005~0.007): (1~5); the mother includes a pregnant mother and / or a lactating mother, and the offspring includes a fetal offspring and / or an infant offspring and an optional childhood offspring. The present invention has found through a large number of studies that a combination of active folic acid substances and N-acetylneuraminic acid supplemented by a pregnant mother and / or a lactating mother can have a positive effect on the memory ability of the offspring.

Description

Technical Field

[0001] The present invention belongs to the field of food, and specifically relates to the use of a nutritional composition for assisting in improving memory ability, and more specifically relates to the use of a nutritional composition containing active folic acid substances and N-acetylneuraminic acid for non-therapeutic purposes of assisting in improving the memory ability of offspring through ingestion by mothers. Background Art

[0002] Neurodevelopment is a complex process that is influenced by the interaction between genetic factors, environmental factors, and experience. The fetal period and infant period are critical periods for brain formation and development, laying the foundation for cognitive, motor, and social-emotional skills from childhood to adulthood. At the same time, the fetal period and infant period are also critical periods for the formation and stereotyping of brain structure and function. They are very sensitive to environmental factors and require adequate nutritional supply. Abnormal changes may lead to long-term irreversible consequences.

[0003] Studies have shown that nutritional status in early life may affect brain development and have a long-term impact on cognitive function, learning and memory ability, causing lasting changes. Nutrients such as protein, long-chain unsaturated fatty acids, choline, and folic acid are essential for brain development by participating in specific biochemical pathways related to the development and function of neurons and glial cells, regulating neuronal migration and proliferation, axon and dendrite growth, synapse formation, myelination, and neuronal apoptosis. Abnormal nutritional status during pregnancy may lead to neurodevelopmental disorders in the offspring after birth. In addition to affecting the basic structure of the offspring's brain tissue, it may also increase the risk of later neurological diseases through epigenetic mechanisms. Reasonable nutrition during pregnancy plays an important role in the cognitive performance of the offspring in the later stage.

[0004] Folic acid is a B vitamin that the human body cannot synthesize on its own and must be obtained through additional intake. As an important cofactor in one-carbon unit metabolism, folic acid mediates the de novo synthesis of purine and thymidine in the body and regenerates homocysteine ​​(Hcy) into methionine, thereby supporting a wide range of methylation reactions in the body, including DNA, protein, neurotransmitters, etc. Folic acid deficiency can lead to impaired DNA synthesis and affect cell division and proliferation. Low folic acid levels during pregnancy are associated with multiple defects in offspring, such as neonatal respiratory distress syndrome (NRDS), congenital heart defects, growth retardation, and low birth weight.

[0005] In the process of folic acid being utilized in the body, methylenetetrahydrofolate reductase (MTHFR) plays an important role, and the gene mutation that determines the synthesis of this enzyme can lead to the occurrence of corresponding diseases. MTHFR C667T has three genotypes: CC, CT and TT, and the corresponding enzyme activities are 100%, 65% and 30%. The TT genotype will greatly reduce the absorption and utilization rate of supplemented synthetic folic acid. 6S-5-Methyltetrahydrofolate, also known as active folic acid, is the most active form of folic acid metabolized after entering the human body. It can be directly absorbed and utilized by the human body, is not affected by the tetrahydrofolate reductase genotype in the body, and will not mask the adverse reactions caused by vitamin B deficiency. It is a better form of folic acid supplementation.

[0006] In order to avoid adverse pregnancy, existing technologies have been used to research and develop nutritional supplements for pregnant women containing folic acid, for example,

[0007] Reference 1 (CN105106217B) discloses a nutritional supplement for pregnant women, comprising L-methylfolate or L-methylfolate, arginine and vitamin B12; wherein L-methylfolate avoids the problem of inability to absorb and convert folic acid due to genetic defects, and avoids the adverse reactions caused by the accumulation of folic acid in the body when pregnant women cannot absorb and convert folic acid; arginine can promote blood circulation and can effectively meet the needs of pregnant women for more semi-essential amino acids during pregnancy; avoid the occurrence of common diseases in pregnant women such as pregnancy-induced hypertension, eclampsia, and pregnancy-induced depression; at the same time, the combination of L-methylfolate and arginine has good fusion.

[0008] N-acetylneuraminic acid, also known as sialic acid, is an acetylated derivative of acidic sugar neuraminic acid. Studies have shown that N-acetylneuraminic acid in breast milk is of great significance in promoting the development of the infant nervous system and enhancing immunity. N-acetylneuraminic acid is involved in cell-to-cell recognition, regulating neurogenesis, cell proliferation and migration, synaptogenesis, cell attachment and axon guidance, and plays a key role in brain information transmission, nerve impulses and synapse formation. Sialic acid is present in high levels in the brain, milk, blood and neural tissue mucin of mammals, with the highest content in the brain. It is an important structural and functional component of gangliosides. Studies have shown that due to the large differences in the content and structure of sialic acid in cow's milk and human milk, breastfed infants have higher overall scores in cognitive development than infants fed with infant formula.

[0009] Currently, there are studies on N-acetylneuraminic acid in brain and neural development related aspects, such as

[0010] Reference 2 (CN110169454A) discloses a children's formula milk powder that is beneficial for protecting eyes and helping brain intelligence development, which includes docosahexaenoic acid, arachidonic acid, lutein, N-acetylneuraminic acid, phosphatidylserine, (3R,3''R)-dihydroxy-β-carotene, phospholipids, vitamins, folic acid, taurine, choline chloride, ferrous sulfate, and zinc sulfate; among them, N-acetylneuraminic acid can promote the formation of synapses and promote nerve development, improve memory, and has antiviral and resistance-enhancing effects.

[0011] Reference 3 (CN102946728B) discloses methods and compositions for improving fetal and child health and development by using, for example, sialic acid for nutritional supplementation, wherein sialic acid is provided to a woman in the form of N-acetylneuraminic acid before, during and / or after pregnancy, and the sialic acid provides developmental benefits to the nervous system and / or brain of the fetus or child of the woman. Summary of the invention

[0012] Problem that the invention aims to solve

[0013] At present, the research focus of nutritional supplements for pregnant women is mainly on how to avoid adverse pregnancies and abnormal fetal development. However, such research can no longer meet the widespread demand in the market development process for nutritional supplements to provide benefits for early human brain development while ensuring the health of mothers and babies.

[0014] Although there are studies on nutrients that can promote brain development and other related aspects, most of these studies start with nutritional interventions in infants and young children. However, studies have shown that even if nutrients are considered to have specific health benefits when given directly to infants and young children, it does not mean that they can play the same role in the development of their offspring when consumed by the mother.

[0015] Although, for example, the above-mentioned reference 3 has studied the developmental benefits of administering sialic acid to women for the nervous system and / or brain of their fetuses or children, it is only a study of sialic acid as a single substance, and has not studied the effects of sialic acid and other possible nutrients co-administered to women on the brain and neural development of their fetuses or children.

[0016] Therefore, there is still room for development of combinations of nutrients that can be ingested by the mother, especially starting from pregnancy, to regulate the maternal nutritional environment and promote aspects related to brain development of her offspring.

[0017] In this regard, the purpose of the present invention is to provide a nutritional composition for non-therapeutic purposes in assisting the improvement of memory ability to help the brain development of offspring. The nutritional composition can advance the nutritional intervention window, improve the maternal nutritional environment during pregnancy, and create better nutritional conditions for aspects related to brain development of offspring while avoiding adverse pregnancy and delivery.

[0018] Solutions for solving problems

[0019] The present invention provides a use of a nutritional composition in preparing a food that is ingested by a pregnant mother and / or a lactating mother to help improve the memory ability of offspring in the fetal period and / or infant period and optionally the offspring in the childhood period.

[0020] Specifically, the present invention provides the following technical solutions:

[0021] [1]. Use of a nutritional composition in preparing a food for improving the memory ability of offspring by ingestion by a mother, wherein the nutritional composition comprises the following essential components: active folic acid and N-acetylneuraminic acid; and, in the nutritional composition, the mass ratio of the active folic acid to the N-acetylneuraminic acid is (0.0005-0.007):(1-5); the mother includes a pregnant mother and / or a lactating mother, and the offspring includes a fetal offspring and / or an infant offspring and optionally a childhood offspring.

[0022] [2] The use according to [1], wherein the active folic acid substance comprises at least one of 6S-5-methyltetrahydrofolate, 6S-5-methyltetrahydrofolate calcium and 6S-5-methyltetrahydrofolate glucosamine salt.

[0023] [3] The use according to [1] or [2], wherein in the nutritional composition, the mass ratio of the active folic acid substance to the N-acetylneuraminic acid is (0.0005-0.007):1.

[0024] [4] The use according to any one of [1] to [3], wherein the food, after being ingested by the mother, helps improve the memory ability of the offspring by maintaining the health of the offspring's nervous system and / or improving the offspring's intestinal flora.

[0025] [5]. The use according to [4], wherein maintaining the health of the offspring's nervous system includes any one or more of reducing nerve cell damage in the offspring's brain tissue, reducing microglial proliferation in the offspring's brain tissue, and reducing inflammatory response in the offspring's brain tissue.

[0026] [6] The use according to [5], wherein reducing the inflammatory response in the offspring brain tissue comprises reducing the content of IL-6 in the offspring brain tissue.

[0027] [7] The use according to any one of [4] to [6], wherein the improvement of the intestinal flora of the offspring comprises increasing the number of Akkermansia genus ( Akkermansia ) and increase the relative abundance of Lactobacillus HT002 in the offspring intestine and reduce the relative abundance of Escherichia-Shigella ( Escherichia-Shigella )'s relative abundance.

[0028] [8] The use according to any one of [1] to [7], wherein the food is candy, beverage, dairy product, baked food, dietary supplement or food for special dietary uses.

[0029] [9]. The use according to any one of [1] to [8], wherein the food is infant food, children's food, adolescent food or adult food; and the adult food is pregnant woman food, postpartum woman food or pregnant and postpartum woman food.

[0030]

[10] . The use according to any one of [1] to [9], wherein the food contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional additives and any acceptable excipients.

[0031] Effects of the Invention

[0032] Through the implementation of the above technical solution, the present invention achieves the following technical effects:

[0033] The present invention has found through a large number of studies that the combination of active folic acid substances and N-acetylneuraminic acid supplemented by pregnant mothers and / or lactating mothers can help maintain the health of the nervous system of their offspring (for example, it can reduce the damage of nerve cells in the brain tissue of the offspring, reduce the proliferation of microglia in the brain tissue of the offspring, and reduce the inflammatory response in the brain tissue of the offspring, etc.) and / or improve the intestinal flora of their offspring (for example, make the intestinal flora of their offspring develop in a beneficial direction), thereby having a positive effect on the memory ability of their offspring. In particular, when pregnant mothers and / or lactating mothers supplement active folic acid substances and N-acetylneuraminic acid in a certain ratio, the above effects are more significant. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1: Results of IBA-1 immunohistochemical detection in the brain tissue of offspring mice; wherein, CK is the blank control group, AL is Example 1, AH is Example 2, BL is Example 3, BH is Example 4, AH+BL is Example 5, and AH+BH is Example 6.

[0035] Figure 2 :Escherichia-Shigella in rat feces under the intervention of different substances Escherichia-Shigella ) change statistical results; wherein, C is the control group, AL is Example 1, AH is Example 2, BL is Example 3, BH is Example 4, AHBL is Example 5, AHBH is Example 6, 1w, 2w, 3w respectively represent 1 week, 2 weeks, 3 weeks after the birth of the offspring.

[0036] Figure 3 : Statistical results of changes in Lactobacillus HT002 genus in feces of offspring mice under the intervention of different substances; wherein, C is the control group, AL is Example 1, AH is Example 2, BL is Example 3, BH is Example 4, AHBL is Example 5, AHBH is Example 6, 1w, 2w, 3w respectively represent 1 week, 2 weeks, and 3 weeks after the offspring mice are born.

[0037] Figure 4 :The fecal Akkermansia spp. in rats under the intervention of different substances Akkermansia ) change statistical results; wherein, C is the control group, AL is Example 1, AH is Example 2, BL is Example 3, BH is Example 4, AHBL is Example 5, AHBH is Example 6, 1w, 2w, 3w respectively represent 1 week, 2 weeks, 3 weeks after the birth of the offspring. DETAILED DESCRIPTION

[0038] 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.

[0039] I. Definition of Terms

[0040] 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.

[0041] In the present invention, the word "may" includes both performing a certain process and not performing a certain process.

[0042] 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.

[0043] 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”.

[0044] 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.

[0045] 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 explicitly 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.

[0046] In the present invention, "normal temperature" refers to an indoor ambient temperature of 23±2°C.

[0047] 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 (ie, liquid obtained directly from the mammary gland) and standardized milk products.

[0048] In the present invention, "gestation period" and "pregnancy period" are used interchangeably and refer to the period from fertilization to birth.

[0049] In the present invention, the "breastfeeding period" refers to the period from the start of breastfeeding to the cessation of breastfeeding after a mother gives birth.

[0050] In the present invention, "exclusively breastfed" means that most of the nutrients and / or energy consumed by the offspring are derived from breast milk.

[0051] In the present invention, the term "primarily fed with breast milk" means that the nutrients and / or energy intake of the offspring mainly (at least 50%, preferably at least 65%, more preferably at least 75%) comes from breast milk.

[0052] In the present invention, "infants" refers to a human group under 36 months of age.

[0053] In the present invention, "infant" refers to a human group under 12 months of age.

[0054] In the present invention, "infants" refers to a human group aged 13 to 36 months.

[0055] In the present invention, "children" refers to a human group that is older than 3 years old and younger than 12 years old and is in the growth and development stage.

[0056] In the present invention, "teenagers" refers to a human group aged 12 years or older and younger than 18 years old.

[0057] In the present invention, "adult" refers to a human group of people aged 18 years or older.

[0058] 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.

[0059] II. Nutritional composition

[0060] The present invention provides a nutritional composition, which comprises the following essential components: active folic acid substances and N-acetylneuraminic acid; and, in the nutritional composition, the mass ratio of the active folic acid substances to the N-acetylneuraminic acid is (0.0005-0.007): (1-5). The active folic acid substances described in the present invention refer to 6S-5-methyltetrahydrofolate and its salts.

[0061] The present invention has found through a large number of studies that supplementing mothers with a combination of active folic acid substances and N-acetylneuraminic acid during pregnancy and / or lactation can help maintain the health of the offspring's nervous system and / or improve the offspring's intestinal flora, thereby assisting in improving the offspring's learning and memory abilities.

[0062] In some embodiments, the main effective ingredients in the nutritional composition are active folic acid substances and N-acetylneuraminic acid, that is, the nutritional composition mainly relies on the active folic acid substances and N-acetylneuraminic acid contained therein to exert specific physiologically active functions, such as assisting in improving the memory ability of fetal offspring and / or infant offspring and optionally childhood offspring through intake by pregnant mothers and / or lactating mothers.

[0063] In some embodiments, the "mother" and "offspring" described in the present invention include mammals, including but not limited to humans, monkeys, gorillas, cows, sheep, cats, dogs, horses, rabbits, mice, rats or guinea pigs, etc.

[0064] In some embodiments, the active folate described in the present invention includes at least one of 6S-5-methyltetrahydrofolate, 6S-5-methyltetrahydrofolate calcium and 6S-5-methyltetrahydrofolate glucosamine salt.

[0065] The present invention does not particularly limit the source of the above-mentioned active folic acid substances. Typically, they can be synthesized by common chemical synthesis methods in the art, such as using folic acid as a raw material, through reduction, methylation, chiral resolution and optional salt formation steps.

[0066] In some specific embodiments, the active folic acid substance described in the present invention is any one of 6S-5-methyltetrahydrofolate, 6S-5-methyltetrahydrofolate calcium and 6S-5-methyltetrahydrofolate glucosamine salt.

[0067] In some more specific embodiments, considering the convenience of obtaining raw materials and the universality of consumption, the active folic acid substance described in the present invention is 6S-5-methyltetrahydrofolate calcium.

[0068] The present invention does not particularly limit the specific source of N-acetylneuraminic acid. Typically, it can be extracted from natural resources containing sialic acid, such as bird's nests, eggs, milk, etc.; or, it can be synthesized by chemical and enzymatic methods, such as condensing N-acetylglucosamine with potassium salt of di-tert-butyloxysuccinic acid, and decarboxylating it under the catalysis of alkali to generate N-acetylneuraminic acid, or producing N-acetylmannosamine by catalyzing N-acetylneuraminic acid aldolase; or, it can be obtained by fermentation with suitable microorganisms such as Escherichia coli and Bacillus subtilis; or, it can be obtained by whole-cell synthesis.

[0069] In some embodiments, the nutritional composition of the present invention consists of active folic acid substances and N-acetylneuraminic acid; and in the nutritional composition, the mass ratio of the active folic acid substances to the N-acetylneuraminic acid is (0.0005~0.007):(1~5).

[0070] In order to obtain a better effect of assisting in improving the memory ability of offspring through intake by pregnant mothers and / or lactating mothers, in some preferred embodiments, in the nutritional composition provided by the present invention, the mass ratio of the active folic acid substance to the N-acetylneuraminic acid is (0.0005-0.007):1; illustratively, in the nutritional composition, the mass ratio of the active folic acid substance to the N-acetylneuraminic acid can be 0.0005:1, 0.0007:1, 0.0009:1, 0.0011:1, 0.0013:1, 0.0015:1, 0.0017:1, 0.0019:1, 0.0021:1, 0.0023:1, 0.0025:1, 0.0027:1, 0.0029:1, 0.0031:1, 0.0033:1, 0.0035:1, 0.0037:1, 0.0039:1, 0.0041:1, 0.0043:1, 0.0045:1, 0.0047:1, 0.0049:1, 0.0051:1, 0.0053:1, 0.0055:1, 0.0057:1, 0.0059:1, 0.0061:1, 0. 0.0063:1, 0.0065:1, 0.0067:1 or 0.0069:1, etc.; preferably, it can be (0.0005-0.006):1; more preferably, it can be (0.0005-0.004):1; even more preferably, it can be (0.0005-0.002):1; further more preferably, it can be (0.001-0.002):1.

[0071] The composition of the present invention is usually an artificially synthesized or compounded composition, that is, it is not a natural composition such as breast milk.

[0072] III. Uses to assist in improving the memory ability of offspring

[0073] The present invention proposes that after active folic acid substances and N-acetylneuraminic acid are compounded, especially after compounding in a specific ratio, they are ingested by pregnant mothers and / or lactating mothers, which helps to maintain the health of the nervous system of the offspring and / or improve the intestinal flora of the offspring, and then can assist in improving the memory ability of the offspring, and there is a synergistic effect between the two substances of active folic acid substances and N-acetylneuraminic acid, which is better than the single active folic acid substances or N-acetylneuraminic acid. At the same time, the present invention believes that the effect of the nutritional composition ingested by the mother to assist in improving the memory ability of the offspring is not only for the fetus present in the mother and the infant who consumes breast milk, but this effect can also continue to the childhood of the offspring and have a lasting effect on the offspring.

[0074] Furthermore, the present invention provides the use of the above-mentioned nutritional composition in preparing food that is ingested by the mother to help improve the memory ability of the offspring, wherein the mother includes a pregnant mother and / or a lactating mother, and the offspring includes fetal offspring and / or infant offspring and optionally childhood offspring.

[0075] In some embodiments, the present invention provides the use of the above nutritional composition in preparing a food for helping to improve the memory ability of offspring ingested by a mother, wherein the mother is a pregnant mother, and the offspring includes fetal offspring and optionally childhood offspring.

[0076] In some embodiments, the present invention provides the use of the above nutritional composition in preparing a food for helping to improve the memory ability of offspring ingested by a mother, wherein the mother is a pregnant mother, and the offspring includes infants and children.

[0077] In some embodiments, the present invention provides the use of the above-mentioned nutritional composition in preparing a food for ingestion by a mother to help improve the memory ability of the offspring, wherein the mother is a pregnant mother, and the offspring include fetal offspring, infant offspring, and optionally childhood offspring.

[0078] In some embodiments, the present invention provides the use of the above nutritional composition in preparing a food for helping to improve the memory ability of offspring ingested by a mother, wherein the mother is a lactating mother, and the offspring includes infants and children.

[0079] In some embodiments, the present invention provides the use of the above-mentioned nutritional composition in preparing a food for ingestion by a mother to help improve the memory ability of the offspring, including pregnant mothers and lactating mothers, and the offspring including fetal offspring and optionally childhood offspring.

[0080] In some embodiments, the present invention provides the use of the above-mentioned nutritional composition in preparing a food for ingestion by a mother to help improve the memory ability of the offspring, including pregnant mothers and lactating mothers, and the offspring including infants and children.

[0081] In some embodiments, the present invention provides the use of the above-mentioned nutritional composition in preparing a food that is ingested by a mother to help improve the memory ability of the offspring, including pregnant mothers and lactating mothers, and the offspring include fetal offspring, infant offspring, and optionally childhood offspring.

[0082] Furthermore, the assistance in improving the memory ability of offspring (including maintaining the health of the offspring's nervous system and improving the offspring's intestinal flora) described in the present invention is not intended to treat or prevent diseases.

[0083] In some embodiments, the infant and toddler offspring of the present invention may be breastfed or not, preferably breastfed. In some embodiments, breastfeeding of the present invention may be completely breastfed, may be mainly breastfed, or may be mixed feeding with a small amount of breast milk and other foods. In some optional embodiments, the offspring of the present invention includes fetal offspring and / or breastfeeding offspring and optional childhood offspring.

[0084] In some embodiments, the food is taken by the mother to help improve the memory of the offspring by maintaining the health of the offspring's nervous system and / or improving the offspring's intestinal flora. In some embodiments, the food is taken by the mother to help improve the memory of the offspring by simultaneously maintaining the health of the offspring's nervous system and improving the offspring's intestinal flora.

[0085] In some specific embodiments, the maintenance of the offspring's nervous system health includes reducing any one or more of nerve cell damage in the offspring's brain tissue, reducing microglial proliferation in the offspring's brain tissue, and reducing inflammatory response in the offspring's brain tissue. In some specific embodiments, the maintenance of the offspring's nervous system health includes reducing nerve cell damage in the offspring's brain tissue, reducing microglial proliferation in the offspring's brain tissue, and reducing inflammatory response in the offspring's brain tissue.

[0086] In some specific embodiments, said reducing the inflammatory response in the offspring brain tissue comprises reducing the level of IL-6 in the offspring brain tissue.

[0087] In some specific embodiments, improving the intestinal flora of the offspring includes increasing the number of Akkermansia genus ( Akkermansia ) and increase the relative abundance of Lactobacillus HT002 in the offspring intestine and reduce the relative abundance of Escherichia-Shigella ( Escherichia-Shigella ) in the intestinal flora of the offspring. In some specific embodiments, the improvement of the intestinal flora of the offspring also includes increasing the relative abundance of Akkermansia ( Akkermansia ) and increase the relative abundance of Lactobacillus HT002 in the offspring intestine and reduce the relative abundance of Escherichia-Shigella ( Escherichia-Shigella )'s relative abundance.

[0088] Studies have shown that Akkermansia Akkermansia) Microorganisms can regulate the immune system and metabolic system through their metabolites such as short-chain fatty acids (SCFAs), amino acids and amino acid derivatives, thereby playing a comprehensive role in the gut-brain axis (Xu R, Zhang Y, Chen S, Zeng Y, Fu X, Chen T, Luo S, Zhang X. The role of the probiotic Akkermansia muciniphila in brain functions: insightsunderpinning therapeutic potential. Crit Rev Microbiol. 2023 Mar;49(2):151-176. doi: 10.1080 / 1040841X.2022.2044286.). Akkermansia spp. ( Akkermansia ) Microorganisms play an important role in a variety of neuropsychiatric diseases related to memory ability (such as Alzheimer's disease, etc.), and have the potential to be a therapeutic target for various neuropsychiatric diseases. At the same time, studies have found that Escherichia-Shigella ( Escherichia-Shigella ) is increased in fecal samples of patients with Alzheimer's disease than in samples of healthy people (Cryan JF, O'Riordan KJ, Sandhu K, Peterson V, Dinan TG. The gut microbiomein neurological disorders. Lancet Neurol. 2020 Feb;19(2):179-194. doi:10.1016 / S1474-4422(19)30356-4. Epub 2019 Nov 18.). In addition, some studies have found that the relative abundance of Lactobacillus HT002 in the intestines of APP / PS1 double transgenic mice is lower than that of wild-type mice (Hu F, Gao Q, Zheng C, Zhang W, Yang Z, Wang S, Zhang Y, Lu T. Encapsulated lactiplantibacillusplantarum improves Alzheimer's symptoms in APP / PS1 mice. J Nanobiotechnology. 2024 Sep 20;22(1):582. doi: 10.1186 / s12951-024-02862-1.)

[0089] The present invention has found that the mother's intake of the above nutritional composition during pregnancy can increase the number of Akkermansia genus ( Akkermansia) and Lactobacillus HT002 and reduced the relative abundance of Escherichia-Shigella ( Escherichia-Shigella Therefore, the present invention believes that by allowing pregnant mothers and / or lactating mothers to supplement the above nutritional composition, it is not only beneficial to the health of the intestinal flora of their offspring, but also can help improve the memory ability of their offspring through the regulation of the gut-brain axis.

[0090] The present invention does not particularly limit the specific food containing the above nutritional composition or the food that can be prepared with the above nutritional composition.

[0091] In some embodiments, at room temperature, the food of the present invention is in the form of liquid or solid.

[0092] In some embodiments, the food of the present invention is candy, such as hard candy, gel candy, crisp candy, compressed candy and aerated candy, etc. In some embodiments, the food of the present invention is beverage, such as carbonated beverage, tea beverage, coffee beverage, fruit and vegetable juice beverage and lactic acid bacteria beverage, etc. In some embodiments, the food of the present invention is dairy products, such as milk powder, cheese, yogurt, liquid milk, etc. In some embodiments, the food of the present invention is baked food, such as bread, cake and biscuits, etc. In some embodiments, the food of the present invention is dietary supplement, such as hard capsule, soft capsule, tablet, oral liquid, pill, granule and powder, etc. In some embodiments, the food of the present invention is special dietary food, such as infant formula, infant supplementary food, supplementary nutritional supplement and special medical formula, etc.

[0093] In some embodiments, the food of the present invention is infant food, children's food, adolescent food or adult food.

[0094] In some preferred embodiments, the food of the present invention is food for pregnant women, food for postpartum women or food for pregnant and postpartum women. The food for pregnant women is suitable for consumption by pregnant women, and the food for postpartum women is suitable for consumption by postpartum women, especially lactating women; preferably, the food is formula milk powder for pregnant women, formula milk powder for postpartum women or oral liquid for pregnant and postpartum women.

[0095] In some embodiments, the food of the present invention is a nutritional composition set for pregnant women or a nutritional composition set for postpartum women.

[0096] In some embodiments, in the food described in the present invention, the mass ratio of the active folic acid substance to N-acetylneuraminic acid is (0.0005~0.007):(1~5).

[0097] The present invention does not particularly limit the specific absolute content of the active folic acid substance and the N-acetylneuraminic acid in food, as long as the requirements of local food-related laws and regulations are met.

[0098] Taking 6S-5-methyltetrahydrofolate calcium and N-acetylneuraminic acid as examples, in some embodiments, the addition of the nutritional composition makes the content of 6S-5-methyltetrahydrofolate calcium in the food, by mass percentage, be 0.0001% to 0.9%, preferably 0.0002% to 0.8%, and more preferably 0.0004% to 0.75%; the content of N-acetylneuraminic acid is 0.05% to 5%, preferably 0.08% to 3%, and more preferably 0.1% to 1%.

[0099] When the mass content of active folic acid substances and N-acetylneuraminic acid in the food is within the above range and is consumed by the mother (e.g., a human (i.e., a pregnant woman) or an animal (i.e., a pregnant female)), it can significantly and especially synergistically improve the brain development of its offspring, and at the same time can also balance the (other) aspects of nutrition required by the human body or animal body (including the mother and the offspring).

[0100] In addition to the components described above for the nutritional composition, the food may also contain other ingredients, such as proteins / amino acids, carbohydrates, fats, vitamins, minerals, etc., which are often contained in formula foods such as maternal formula foods such as milk powder or beverages, candies, etc.

[0101] In addition, depending on the type of food and the final needs of the applicable objects, in some embodiments, the food 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.

[0102] 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; fruit and vegetable substances such as onions, cucumbers, tomatoes, cauliflower, 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 medicinal and edible.

[0103] As for animal dairy ingredients, examples include fresh milk from cows, sheep, etc., as well as reprocessed dairy products such as whole milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, hydrolyzed whey protein powder, and casein powder.

[0104] Examples of animal meat ingredients include meat ingredients from pigs, cattle, sheep, aquatic animals or poultry.

[0105] Functional added ingredients include vitamin supplements, mineral supplements, nucleotide supplements, dietary fiber, functional polyunsaturated fatty acid supplements, and the like.

[0106] 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. Example

[0107] 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 conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the materials or instruments used are all conventional products that can be obtained commercially.

[0108] Experimental animals and groups used in the experiment

[0109] 70 8-week-old sexually mature SPF female rats and 70 SPF male rats were selected; purchased from Hangzhou Medical College, animal certificate number 20240318Aazz0100000836, production license number: SCXK (Zhejiang) 2024-002. Rearing conditions: Animals were raised in a barrier environment, with an indoor temperature of 25±1℃, humidity of 45±5%, free access to drinking water, and 12h light and dark alternation every day. This experiment passed the animal experiment ethics review of Southeast University, ethics number: 20240316003.

[0110] After 3 days of adaptive feeding, female and male rats were placed in the same cage at a ratio of 1:1. The intervention started when the mother rat showed vaginal plugs, and the mother rats were continuously gavaged until the offspring rats were weaned. The experimental group was divided into groups and given the corresponding dose of sample aqueous solution, and the control group was given the corresponding dose of normal saline, gavaged once a day for 6 consecutive weeks, and each dose group was given maintenance feed. The low and high doses of active folic acid substances were 5.3 and 10.1 (μg / d / rat), and the low and high doses of N-acetylneuraminic acid were 1.5 and 7.5 (mg / d / rat). The specific doses of each group are shown in Table 1 below. During the gavage process, the brain development-related indicators of the offspring were tested.

[0111] Table 1 Oral dose of experimental animals

[0112]

[0113] Instruments, consumables and reagents used in the experiment

[0114] The consumables, reagents, and experimental instruments used in the experiment are shown in Tables 2 and 3.

[0115] Table 2 Main experimental consumables and reagents

[0116]

[0117] Table 3 Main experimental instruments

[0118]

[0119] Experimental Example 1: Nutrients improve the learning and memory abilities of offspring mice

[0120] The Morries water maze experiment is mainly used to test the learning and memory ability of experimental animals for spatial position and orientation (spatial positioning). It is widely used in scientific research fields such as learning and memory, intelligence and aging. It is widely recognized in the world and is a classic experiment in behavioral science, especially learning and memory research. The specific experimental method is as follows.

[0121] 1. Positioning navigation experiment

[0122] After the offspring were separated from the mother and weaned, they were placed in a pool to swim freely for 2 minutes to familiarize themselves with the maze environment. The training was carried out 4 times in a fixed time period every day. At the beginning of the training, the platform was placed in the first quadrant, and the offspring was placed in the pool facing the pool wall from any of the four starting points on the pool wall. The time it took for the offspring to find the platform (escape latency) and the swimming path were recorded. After the offspring found the platform or failed to find the platform within 90 seconds, the experimenter took it to the platform and rested on the platform for 15 seconds before the next experiment. The average value of the offspring's 4 training latencies was used as the offspring's learning performance for the day. The directional navigation experiment lasted for 4 days.

[0123] 2. Space Exploration Experiment

[0124] On the fifth day, the platform was removed at the same time, and the offspring was placed in the water at a fixed point in the fourth quadrant. The video recorded the time the offspring stayed in the quadrant where the original platform was located and the number of times it passed the original platform position within 60 seconds. If the animal stayed in the quadrant where the original platform was located for a longer time and passed the original platform position more times, it means that it has a good long-term memory of the platform position.

[0125] 3. Other requirements during the experiment

[0126] Keep the water temperature and environmental conditions in the maze stable, conduct training at a fixed time every day, keep the light and reference signs around the maze clear and unchanged, and at the end of each day's experiment, wipe off the moisture on the surface of the mice, replace the bedding to keep the breeding environment dry. At the same time, the mice are free to wade in water and eat, and are raised in a barrier environment.

[0127] 4. Water maze experiment data analysis

[0128] Super Maze software was used to process the video files of the water maze test and to record and analyze the swimming trajectories of the water maze test mice.

[0129] 5. Experimental Results

[0130] Table 4 Number of times the rats crossed the platform area (times)

[0131]

[0132] The more times the animal passes through the original platform position, the better its spatial memory of the platform position. The results in Table 4 show that compared with the control group, the number of times the offspring of Examples 1, 2, 3, 4, 5, and 6 cross the platform area increases to varying degrees; among them, the number of times the offspring of Examples 5 and 6 cross the platform area increases to varying degrees compared with Examples 2, 3, and 4.

[0133] The results showed that compared with the blank control group, the number of platform area crossings of the offspring mice in the groups gavage with different doses of active folic acid substances alone and different doses of N-acetylneuraminic acid alone increased, and showed an increasing trend with increasing doses; it was unexpectedly found that the number of platform area crossings of the offspring mice after gavage with the combination of active folic acid substances and N-acetylneuraminic acid was higher than that of the single gavage, and the combination of high doses of active folic acid substances and high doses of N-acetylneuraminic acid was more effective in promoting spatial memory of offspring mice.

[0134] Table 5 Platform latency of offspring mice (s)

[0135]

[0136] The shorter the animal's platform latency is, the better its long-term memory of the platform location is. The results in Table 5 show that, relative to the control group, the platform latency of the offspring mice in Examples 1, 2, 3, 4, 5, and 6 are shortened to varying degrees; among them, the platform latency of the offspring mice in Examples 5 and 6 is shortened to varying degrees compared with that in Examples 2, 3, and 4, and the platform latency of the offspring mice in Example 6 is shorter than that in Example 5.

[0137] The results showed that compared with the blank control group, the platform latency of the offspring mice was shortened in the groups with different doses of active folic acid substances and different doses of N-acetylneuraminic acid given by oral gavage alone, and the N-acetylneuraminic acid dose group showed a trend of increasing effect with increasing dose; it was unexpectedly found that the platform latency of the offspring mice after oral gavage of the combination of active folic acid substances and N-acetylneuraminic acid was lower than that of the single gavage, and the combination of high doses of active folic acid substances and high doses of N-acetylneuraminic acid was more effective in promoting the long-term memory of offspring mice.

[0138] Table 6 The time that the young mice stayed in the platform area (s)

[0139]

[0140] The longer the animal stays in the original platform area, the better its long-term memory of the platform location. The results in Table 6 show that, compared with the control group, the stay time of the offspring mice in Examples 1, 2, 3, 4, 5, and 6 in the platform area increased to varying degrees; among them, the stay time of the offspring mice in Example 6 in the platform area increased to varying degrees compared with Examples 2 and 4.

[0141] The results showed that compared with the blank control group, the time that the offspring mice stayed in the platform area was increased in the groups that were gavaged with different doses of active folic acid substances alone and different doses of N-acetylneuraminic acid alone. It was unexpectedly found that the time that the offspring mice stayed in the platform area after gavage with the combination of high doses of active folic acid substances and high doses of N-acetylneuraminic acid was higher than that of the single gavage, indicating that the combination of high doses of active folic acid substances and high doses of N-acetylneuraminic acid is more effective in promoting the long-term memory of offspring mice.

[0142] Experimental Example 2: Nutrients maintain the health of the nervous system of offspring mice

[0143] After the brain tissue was thawed, it was rinsed in 4°C precooled saline solution to remove blood. After the residual saline solution in the brain tissue was absorbed with filter paper, a sample was accurately weighed, and PBS solution was added at a ratio of 1:10 (0.1 g of tissue was added to 0.9 mL of PBS solution). A 10% homogenate was prepared using a homogenizer, and the homogenate was used to detect the content of neural development-related proteins and inflammatory factors in the brain tissue.

[0144] 1. Neuron-specific enolase (NSE) detection in the brain tissue of offspring mice: The NSE content in the brain tissue of offspring mice was detected using a fully automatic microplate reader with reference to the NSE enzyme-linked immunosorbent Elisa method.

[0145] 2. Detection of ionized calcium binding adaptor molecule-1 (IBA-1) in the brain tissue of offspring mice: Use an automatic microplate reader to refer to the IBA-1 enzyme-linked immunosorbent assay (ELISA) method to detect the IBA-1 content in the brain tissue of offspring mice.

[0146] 3. IBA-1 immunohistochemical detection of brain tissue of offspring mice:

[0147] Table 7 Antibody information and repair conditions

[0148]

[0149] a. Dewax the paraffin sections to make them hydrated: sequentially place the sections in environmentally friendly dewaxing solution I for 10 min, environmentally friendly dewaxing solution II for 10 min, environmentally friendly dewaxing solution III for 10 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, and then wash with distilled water.

[0150] b. Antigen repair: See Table 7 for repair conditions. During the repair process, prevent excessive evaporation of the buffer and do not allow the slides to dry out. After repair, cool naturally. Place the slides in PBS (pH 7.4) and wash on a decolorizing shaker 3 times, 5 minutes each time.

[0151] c. Circle serum blocking: After the slices are slightly dried, use a histochemical pen to draw a circle around the tissue, add BSA (if the primary antibody is from goat, use 10% donkey serum to block, and if the primary antibody is from other sources, use 3% BSA to block), and block for 30 minutes.

[0152] d. Add primary antibody: Add the prepared primary antibody dropwise and place the slices flat in a humidified box and incubate at 4°C overnight.

[0153] e. Add secondary antibody: Place the slide in PBS (pH 7.4) and wash on a decolorizing shaker for 3 times, 5 minutes each time. Add the corresponding secondary antibody and incubate at room temperature for 50 minutes in the dark.

[0154] f. DAPI counterstaining of cell nuclei: Place the slide in PBS (pH 7.4) and wash on a decolorizing shaker for 3 times, 5 minutes each time. Add DAPI staining solution and incubate at room temperature for 10 minutes in the dark.

[0155] g. Quench tissue autofluorescence: Place the slide in PBS (pH 7.4) and wash on a decolorizing shaker 3 times, 5 minutes each time. Add autofluorescence quencher B solution for 5 minutes, and rinse with running water for 10 minutes. (If you need to observe tissue autofluorescence, remove this step.)

[0156] h. Sealing: Sealing with anti-fluorescence fading mounting medium.

[0157] i. Image acquisition: DAPI excitation wavelength 330-380nm, emission wavelength 420nm; 488 excitation wavelength 465-495nm, emission wavelength 515-555nm; CY3 excitation wavelength 510-560nm, emission wavelength 590nm; CY5 excitation wavelength 608-648nm, emission wavelength 672-712nm.

[0158] j. Aipathwell® software was used to automatically identify and locate the nucleus of DAPI blue fluorescence and expand the cytoplasm; the single-color channel fluorescence photos were reversely decolorized (positive signal was black and background was white), the black positive signal was read and the number of positive cells and area, cumulative optical density value IOD (integrated optical density), tissue area and other parameters were calculated.

[0159] 4. Detection of interleukin-6 (IL-6) in the brain tissue of offspring mice: Use a fully automatic microplate reader to refer to the IL-6 enzyme-linked immunosorbent Elisa method to detect the IL-6 content in the brain tissue of offspring mice.

[0160] 5. Experimental Results

[0161] Table 8 NSE detection results of brain tissue of offspring mice (pg / mg·prot)

[0162]

[0163] NSE is an enzyme specific to neurons and neuroendocrine cells, and is a sensitive indicator for evaluating the degree of nerve cell damage and judging the prognosis. The results in Table 8 show that, compared with the control group, the NSE values ​​of the brain tissues of the offspring mice of Examples 2, 3, 5, and 6 were all reduced to varying degrees; the NSE value of the brain tissues of the offspring mice of Example 6 was reduced to varying degrees compared with Examples 2 and 4, and the NSE value of the brain tissues of the offspring mice of Example 5 was reduced to varying degrees compared with Examples 2 and 3.

[0164] The results showed that compared with the blank control group, there was no significant difference in the NSE value of the brain tissue of offspring mice between the groups that were gavaged with different doses of active folic acid substances alone and the groups that were gavaged with different doses of N-acetylneuraminic acid alone. However, it was unexpectedly found that the NSE value of the brain tissue of offspring mice after gavage with the combination of active folic acid substances and N-acetylneuraminic acid was lower than that of the single gavage, indicating that the combination of N-acetylneuraminic acid and active folic acid substances can achieve a better effect in reducing nerve cell damage.

[0165] Table 9 IBA-1 detection results of offspring mouse brain tissue (pg / mg·prot)

[0166]

[0167] IBA-1 is specifically expressed in microglia of the central nervous system, and reducing microglial proliferation can reduce related neuritis and synaptic damage. The results in Table 9 show that, compared with the control group, the IBA-1 values ​​of the brain tissues of the offspring mice of Examples 2, 3, 4, 5, and 6 were all reduced to varying degrees; among them, the IBA-1 values ​​of the brain tissues of the offspring mice of Example 6 were reduced to varying degrees compared with Examples 2 and 4.

[0168] The results showed that compared with the blank control group, the IBA-1 values ​​in the brain tissue of offspring mice in the group gavage with high-dose active folic acid substances alone and different doses of N-acetylneuraminic acid alone were reduced to varying degrees; but it was unexpectedly found that the IBA-1 value in the brain tissue of offspring mice after gavage with the combination of high-dose active folic acid substances and high-dose N-acetylneuraminic acid was lower than the effect of gavage alone, indicating that the combination of high-dose active folic acid substances and high-dose N-acetylneuraminic acid can obtain a better effect of reducing microglial proliferation.

[0169] The results of IBA-1 immunohistochemistry test of offspring rat brain tissue showed that compared with the control group, the number and area of ​​IBA-1 labeled microglia staining in offspring rat brain tissue of Example 2, 3, and 6 groups were lower, among which the number and area of ​​IBA-1 labeled microglia staining in offspring rat brain tissue after intervention with the composition of Example 6 were lower than those in Example 2 and Example 3, indicating that a certain dose of active folic acid substances and N-acetylneuraminic acid gavage to mother rats can reduce the activation of microglia in offspring rat brain tissue. Therefore. It was unexpectedly found that the number and area of ​​IBA-1 labeled microglia staining in offspring rat brain tissue after gavage of a high dose of active folic acid substances plus a high dose of N-acetylneuraminic acid were lower than the effect of gavage alone, indicating that a better effect of reducing microglial proliferation can be obtained by combining a high dose of active folic acid substances and a high dose of N-acetylneuraminic acid.

[0170] Table 10 IL-6 detection results in brain tissue of offspring mice (pg / mg·prot)

[0171]

[0172] IL-6 is synthesized by neurons and glial cells, and has both neuroprotective and neurotoxic effects during brain injury. It protects or promotes nerve repair at low normal physiological concentrations or levels, but can cause neuronal damage when highly expressed. The results in Table 10 show that, relative to the control group, the IL-6 values ​​in the brain tissues of the offspring mice of Examples 1, 2, 4, 5, and 6 were all reduced to varying degrees; among them, the IL-6 values ​​in the brain tissues of the offspring mice of Example 6 were reduced to a certain extent compared to Examples 2 and 4.

[0173] The results showed that compared with the blank control group, the IL-6 values ​​in the brain tissue of offspring mice in the groups that were gavaged with different doses of active folic acid substances alone and the group that were gavaged with a high dose of N-acetylneuraminic acid alone were reduced to varying degrees; but it was unexpectedly found that the IL-6 values ​​in the brain tissue of offspring mice after gavage with the combination of high doses of active folic acid substances and high doses of N-acetylneuraminic acid were significantly lower than the effect of gavage alone, indicating that the combination of high doses of active folic acid substances and high doses of N-acetylneuraminic acid can achieve a better effect in reducing the inflammatory response in brain tissue.

[0174] Experimental Example 3: Nutrients improve the intestinal flora of offspring mice

[0175] High-throughput sequencing of bacteria in fecal samples of mother mice and offspring using 16S rDNA:

[0176] 1) Extraction of total DNA of the microbiome: The total DNA of the microbiome in feces was extracted by the cetyltrimethylammonium bromide method (CTAB method), and the quality of DNA extraction was tested by agarose gel electrophoresis. The DNA was quantified by ultraviolet spectrophotometer.

[0177] 2) PCR amplification: The V3~V4 region of bacterial 16s rDNA was selected for gene amplification and sequencing. The primer sequences are shown in Table 11 below, the PCR reaction system is shown in Table 12 below, and the PCR reaction conditions are shown in Table 13 below.

[0178] Table 11 Primer sequences

[0179]

[0180] Table 12 Reaction system

[0181]

[0182] Table 13 Reaction conditions

[0183]

[0184] 3) PCR products were purified by AMPure XT beads (Beckman Coulter Genomice, MA, USA) and quantified by Qubit (Invitrogen, USA).

[0185] 4) The purified PCR products were evaluated using the Agilent 2100 bioanalyzer (Agilent, USA) and the Illumina (Kapa Biosciences, Wobum, MA, USA) library quantification kit. The qualified library concentration was above 2 nM. The qualified libraries for sequencing were gradient diluted, mixed according to the required sequencing amount, and denatured to single strands by NaOH for sequencing. The NovaSeq 6000 sequencer was used for 2×250 bp double-end sequencing, and the corresponding reagent was the NovaSeq 6000 SP Reagent Kit (500 cycles).

[0186] 5) For the paired-end data obtained by sequencing, the sample data is first split according to the Barcode information, and the adapter and Barcode sequence are removed, and then the sequence is spliced ​​to obtain the optimized sequence. After removing the chimera sequence, OUT cluster analysis is performed, and taxonomic analysis is performed on the representative sequence of OUT. Based on the results of OUT cluster analysis, multiple diversity index analyses and sequence depth detection are performed on OUT; based on taxonomic information, colony structure statistical analysis is performed at each taxonomic level.

[0187] The levels of intestinal flora in the feces of female mice were analyzed. The levels of Lactobacillus genus ( Ligilactobacillus ) had no significant changes, but Akkermansia spp. ( Akkermansia ) relative content decreased. After intervention with active folic acid and N-acetylneuraminic acid, the intestinal flora of the mother mice showed significant changes, specifically, the lactic acid bacteria genus ( Ligilactobacillus ) showed a decreasing trend in most intervention groups; while in most intervention groups, the relative content of Akkermansia ( Akkermansia ) increased in relative abundance, and the relative contents were higher than those in the control group, among which the high and low dose groups of active folic acid substances and the high and low dose groups of N-acetylneuraminic acid showed an obvious upward trend. At the end of the experiment, the control group had Akkermansia genus ( Akkermansia ) relative abundance was significantly reduced, and the intestinal Akkermansia genus ( Akkermansia The results showed that the intake of a certain dose of active folic acid and N-acetylneuraminic acid during pregnancy can promote the increase of the relative content of beneficial bacteria in the intestines of maternal mice.

[0188] Escherichia-Shigella in the intestines of offspring mice Escherichia-Shigella ) level as Figure 2 As shown, 7 days after birth, the intestinal flora of mice in each group contained Escherichia-Shigella ( Escherichia-Shigella ) content was high, belonging to the dominant flora. After continuous breastfeeding for 3 weeks, except for Example 1, the Escherichia-Shigella genus ( Escherichia-Shigella ) were lower in relative abundance than those in the control group.

[0189] The level of Lactobacillus HT002 in the intestine of offspring mice Figure 3 As shown, 7 days after the birth of the offspring, the relative abundance of Lactobacillus HT002 in the intestinal flora of the offspring of the mice in each group was different. Except for Example 1, the relative abundance of Lactobacillus HT002 in other groups was lower than that in the control group. After continuous breastfeeding for 3 weeks, the relative abundance of Lactobacillus HT002 in the offspring of the mice in each group increased, and the relative abundance of Lactobacillus HT002 in the offspring of the mice in Examples 2, 3, and 6 was higher than that in the control group.

[0190] Akkermansia spp. in the intestines of offspring mice Akkermansia ) level as Figure 4 As shown, 7 days after the birth of the offspring, the intestinal flora of Akkermansia genus ( Akkermansia ) had little difference in relative abundance; after continuous breastfeeding for 3 weeks, compared with the control group, the number of Akkermansia genus ( Akkermansia ) have increased in relative abundance to varying degrees.

[0191] The above results show that the intake of active folic acid and N-acetylneuraminic acid by mother mice starting from pregnancy can promote the content of beneficial bacteria in the intestines of offspring mice, which in turn can improve the memory ability of offspring mice through regulating the gut-brain axis.

Claims

1. Use of a nutritional composition in preparing a food for ingestion by a mother to assist in improving the memory ability of offspring, characterized in that: The nutritional composition comprises the following essential components: active folic acid substances and N-acetylneuraminic acid; and, in the nutritional composition, the mass ratio of the active folic acid substances to the N-acetylneuraminic acid is (0.0005-0.007):(1-5); the mother comprises a pregnant mother and / or a lactating mother, and the offspring comprises a fetal offspring and / or an infant offspring; After being ingested by the mother, the food helps improve the memory ability of the offspring by maintaining the health of the offspring's nervous system and improving the offspring's intestinal flora; The maintaining of the nervous system health of the offspring includes any one or more of reducing the damage of nerve cells in the brain tissue of the offspring, reducing the proliferation of microglia in the brain tissue of the offspring, and reducing the inflammatory response in the brain tissue of the offspring; The improving of the intestinal flora of the offspring includes increasing the number of Akkermansia genus ( Akkermansia ) and increase the relative abundance of Lactobacillus HT002 in the offspring intestine and reduce the relative abundance of Escherichia-Shigella ( Escherichia-Shigella )'s relative abundance; The active folic acid substance includes at least one of 6S-5-methyltetrahydrofolate, 6S-5-methyltetrahydrofolate calcium and 6S-5-methyltetrahydrofolate glucosamine salt.

2. The use according to claim 1, characterized in that In the nutritional composition, the mass ratio of the active folic acid substance to the N-acetylneuraminic acid is (0.0005-0.007):

1.

3. The use according to claim 1, characterized in that The reducing the inflammatory response in the brain tissue of the offspring includes reducing the content of IL-6 in the brain tissue of the offspring.

4. The use according to claim 1 or 2, characterized in that: The food is candy, beverage, dairy product, bakery product, dietary supplement or food for special dietary use.

5. The use according to claim 1 or 2, characterized in that: The food is food for pregnant women or food for postpartum women.

6. The use according to claim 1 or 2, characterized in that: The food is food for pregnant women.

7. The use according to claim 1 or 2, characterized in that: The food 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 auxiliary materials.

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