Application of sialic acid in preparation of products for promoting growth

Abstract

The invention provides application of sialic acid in preparation of a product for promoting growth, and relates to the technical field of prebiotics. The time period of oral sialic acid, the main body of oral sialic acid, the dosage of oral sialic acid and the specific influence of oral sialic acid on intestinal-brain co-development are systematically studied. The invention discloses the effect of early-stage supplement of sialic acid on promotion of early-stage tibia growth and intestine-brain co-development for the first time.

Description

Technical Field

[0001] The present application relates to the technical field of prebiotics, and in particular to the application of sialic acid in the preparation of growth-promoting products. Background Art

[0002] The early years of life are also a crucial stage for the co-development of the gut and brain. The brain possesses cognitive abilities, enabling complex working memory, self-awareness, social interaction, tool-making, and cultural development. The development of the fine structure of the adult brain is the result of embryonic stem cells undergoing precise gene expression regulation and cell signaling, which then differentiate into more sophisticated cells and migrate to their sites of action. Rapid structural changes occur in the early years of life, including an increase in size, the formation and differentiation of synapses, myelination, and a rapid improvement in cognitive functions such as hearing, vision, perception, and memory. Simultaneously, the development of the gut and brain occurs with a certain degree of synchronization. Increases in the abundance and diversity of intestinal microbes, maturation of intestinal tissue (such as changes in crypt and villus structure), and maturation of the enteric nervous system (such as changes in nerve density and neuronal types) also occur rapidly during the first three years of life. The gut is also the body's largest immune organ, accounting for 70% of the body's total immune system. Over 95% of infectious diseases are associated with the digestive tract. The intestinal microbiota of infants and young children is unstable and easily affected by environmental factors and dietary habits, leading to unstable intestinal function. An unhealthy gut can lead to a variety of illnesses, such as colds, fevers, and indigestion. In severe cases, it can even affect the growth and development of infants and young children. The development of the gut and brain is mediated by a complex bidirectional communication channel called the gut-brain axis (GBA), which is jointly constructed by the nervous system, endocrine system, gut microbiota, immune system, and metabolic mediators. Through this channel, gut and brain development mutually influence each other, jointly promoting the healthy growth of infants and young children. The early colonization of the gut microbiota is influenced by a variety of prenatal and postnatal factors. A growing body of research has found that the development of the gut microbiota is closely linked to early nervous system development and may have a profound impact on this development. Metabolites such as short-chain fatty acids produced by the gut microbiota can enter the brain through the bloodstream, affecting neuronal growth, differentiation, and synaptic formation, thereby influencing brain development. The gut microbiota can also influence brain function and development by producing signaling molecules such as neurotransmitters and neuropeptides. Furthermore, specific bacterial genera influence brain development by influencing the state of the immune system. The gut microbiota also participates in regulating the function of the blood-brain barrier, thereby protecting the brain from harmful substances. An imbalance in the intestinal flora can disrupt the integrity of the blood-brain barrier, allowing harmful substances to enter the brain and affect the development of the nervous system. Therefore, protecting the gut-brain development of infants and young children in the early stages of life is of great significance.

[0003] Sialic acid (NANA), also known as N-acetylneuraminic acid, acetylenurosyl acid, and N-acetylmannosaminepyruvic acid, CAS number: 131-48-6, is a naturally occurring carbohydrate. It was originally isolated from submandibular gland mucin, hence its name. Sialic acid typically exists as oligosaccharides, glycolipids, or glycoproteins. Sialic acid is present in key tissues and plays important roles. The brain contains more sialic acid than any other tissue in the body, 75% of which is bound to gangliosides, with some existing in free form. Sialic acid is synthesized in the cytoplasm and transported to the nucleus, where it is activated to form the active nucleotide donor CMP-Sia. CMP-Sia returns to the cytoplasm and is then transported to the Golgi apparatus, where it is catalyzed for transfer to endogenous receptors. Therefore, sialic acid participates in the movement and release of neurotransmitters in the brain, alters synaptic structure, and promotes neuronal differentiation, development, and regeneration.

[0004] Sialic acid in the body is mainly synthesized in the liver. Infants' early self-synthesis of sialic acid is insufficient to meet the demand for sialic acid for brain development, so exogenous intake is required. For infants and young children, breast milk is an important source of sialic acid, but due to practical factors, many infants and young children cannot be fully breastfed after birth. Therefore, it is of great significance to explore various ways to supplement sialic acid exogenously. Current technologies are mostly developed around sialic acid nutritional supplements for infants and young children, but the form is relatively simple and lacks attention to the coordinated construction of the gut and brain during critical periods. Summary of the Invention

[0005] To address the above issues, the present invention aims to address current technical challenges regarding the use of sialic acid in early life. One objective is to identify an appropriate oral dosage. A second objective is to develop early intervention methods suitable for both mother and infant. A third objective is to demonstrate the potential of sialic acid to promote both early tibial growth and gut-brain development.

[0006] In one aspect, the present application provides the use of sialic acid in preparing a product for promoting height growth.

[0007] Furthermore, the promoting of height growth includes promoting an increase in bone length; preferably, the promoting of height growth includes promoting the growth of the tibia.

[0008] Furthermore, the sialic acid includes one or more of N-acetylneuraminic acid (N-acetylneuraminic acid, Neu5Ac), N-glycoulylneuraminic acid (Neu5Gc), and deaminoneuraminic acid (Deaminoneuraminic acid, KDN);

[0009] Furthermore, the sialic acid is N-acetylneuraminic acid (N-acetylneuraminic acid, Neu5Ac).

[0010] This application first discovered that sialic acid can promote tibial growth, thereby promoting height growth and weight growth.

[0011] Furthermore, the application at least includes regulating the intestinal microenvironment; preferably, regulating the intestinal microenvironment includes reducing the abundance of harmful microorganisms and / or increasing the abundance of beneficial microorganisms; more preferably, the beneficial microorganisms include Bacteroides strains.

[0012] First, some metabolites of beneficial intestinal flora participate in the regulation of the endocrine axis, and promote individual growth by regulating the levels of multiple hormones. Second, height growth depends on adequate nutrition (such as protein, calcium, vitamin D, zinc, etc.), and intestinal flora directly affects the digestion and absorption of nutrients. It can participate in the synthesis of B vitamins and K2, thereby activating osteocalcin; the metabolites of intestinal flora (such as propionic acid, butyric acid, etc.) can promote the absorption of minerals. In addition, intestinal flora can also affect the hypothalamus and liver through the vagus nerve system, regulating the secretion of growth hormone GH and insulin-like growth factor IGF-1. Third, intestinal microbial flora is also particularly important for the normal development of the intestinal barrier in young children. Long-term exposure to intestinal pathogens in early life can lead to the destruction of intestinal structure and function, which in turn causes problems such as environmental intestinal dysfunction, and thus affects individual development.

[0013] Furthermore, the application at least includes promoting the growth of intestinal cells; preferably, at least includes promoting the proliferation of colon goblet cells.

[0014] The growth of intestinal cells determines an individual's absorption and metabolism of nutrients. Therefore, the proliferation and health of intestinal cells are closely related to an individual's height growth. The growth and functional status of intestinal cells have an important impact on height growth, mainly through mechanisms such as maintaining the integrity of the intestinal barrier, regulating immune balance, and affecting nutrient absorption. Among them, goblet cells are specialized cells of the intestinal epithelium that can secrete mucus to reduce intestinal inflammation and leakage, ensuring nutrient absorption; the mucus layer also provides a habitat for probiotics and inhibits the colonization of harmful bacteria. Goblet cells also affect local immune responses by secreting mucins (such as MUC2), and chronic low-grade inflammation inhibits the activity of the growth hormone (GH)-IGF-1 axis, which is not conducive to growth and development.

[0015] Furthermore, the use at least includes improving neuronal plasticity; preferably, improving neuronal plasticity includes regulating the level of neural cell adhesion molecule (NCAM1); preferably, improving neuronal plasticity includes increasing the level of neural cell adhesion molecule (NCAM1).

[0016] Neuronal plasticity does not directly regulate growth-related bone cell division, but indirectly affects bone development by regulating neuroendocrine signals. Neuronal plasticity is the ability of the nervous system to adapt to environmental changes through mechanisms such as synaptic plasticity, neurogenesis, and neural network reorganization. Neuronal plasticity can participate in regulating the activity of the hypothalamus-pituitary-growth axis (GH-IGF-1 axis) and affect the secretion of growth hormone; it can also perform neural integration of movement and mechanical stimulation. The plasticity of the cerebellum and motor cortex optimizes motor skills (such as jumping and stretching), increases mechanical stress on bones, and stimulates osteoblast activity; neuronal plasticity is also the neural basis of sleep quality, and deep sleep is highly synchronized with the pulsed secretion of GH.

[0017] The product of the present application may further contain excipients, which may be appropriate solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, etc. Preferably, the product is an oral preparation.

[0018] Oral preparations containing sialic acid can be foods, medicines, health products, etc., specifically in the form of solid beverages, oral liquids, soft candies, etc. Those skilled in the art can select excipients and conventional preparation methods according to the dosage form of the preparation, and this application does not impose specific restrictions.

[0019] In a preferred embodiment, a sialic acid solid beverage comprises: 0.2-0.3 parts of sialic acid, 1-10 parts of blueberry powder, 10-20 parts of oligofructose, 10-20 parts of maltodextrin, and 0.1-0.5 parts of zinc gluconate. The preparation method of the solid beverage comprises: preparing the components into powders and mixing them uniformly.

[0020] In a preferred embodiment, a sialic acid oral liquid comprises: 0.2-0.3 parts sialic acid, 10-20 parts potassium sorbate, 10-20 parts isomalto-oligosaccharide, 1-10 parts citric acid, and 50-200 parts purified water. The preparation method comprises: dissolving the components in water, filtering, sterilizing, and canning.

[0021] In a preferred embodiment, a sialic acid soft candy comprises: 10-50 parts concentrated fruit juice, 0.2-0.3 parts sialic acid, 1-5 parts oligofructose, 1-5 parts isomalto-oligosaccharide, 0.1-0.5 parts citric acid, 10-50 parts white sugar, 1-5 parts carrageenan, and 1-5 parts pectin. The preparation method comprises: preparing a mixed gel solution according to a conventional soft candy making method, casting and molding, and packaging to obtain the sialic acid soft candy.

[0022] Furthermore, the concentration of sialic acid is at least 3 mg / kg or above; preferably, 5 mg / kg-500 mg / kg.

[0023] The concentration of sialic acid may be any one of 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 200 mg / kg, 300 mg / kg, 400 mg / kg, and 500 mg / kg.

[0024] In another aspect, the present application provides a method for promoting height growth and / or promoting brain-gut co-development, comprising administering sialic acid to an individual.

[0025] Furthermore, the concentration of sialic acid is at least 3 mg / kg or above; preferably, 5 mg / kg-500 mg / kg.

[0026] The concentration of sialic acid may be any one of 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 200 mg / kg, 300 mg / kg, 400 mg / kg, and 500 mg / kg.

[0027] It will be understood by those skilled in the art that the specific dosage can be adjusted according to individual circumstances, and therefore is not specifically limited here.

[0028] Preferably, the individual is a mammal; more preferably, the mammal is a mouse and / or a human.

[0029] Furthermore, the method of administration includes oral administration.

[0030] Those skilled in the art can prepare sialic acid into common oral preparations for use.

[0031] Furthermore, the method of administration at least comprises taking a product containing sialic acid during pregnancy and / or lactation.

[0032] The cycle of use during pregnancy can be 1-40 weeks.

[0033] During pregnancy, the cycle can be 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, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 weeks.

[0034] The duration of use during breastfeeding can be 1-104 weeks.

[0035] The dosage cycle during breastfeeding can be any number of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 51, 52, 53, 60, 70, 80, 90, 100, 101, 102, 103, or 104 weeks.

[0036] Furthermore, the method of administration at least comprises administering a product containing sialic acid to an individual after birth.

[0037] Preferably, the individual consumes the sialic acid-containing product for 1-104 weeks after birth.

[0038] More preferably, the individual consumes the product containing sialic acid for 3-6 weeks after birth.

[0039] The dosage cycle can be any number of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 51, 52, 53, 60, 70, 80, 90, 100, 101, 102, 103, or 104 weeks.

[0040] In a preferred embodiment, a method for promoting height growth and / or promoting brain-gut co-development comprises any one or more of the following A1)-A3):

[0041] A1) Oral administration of sialic acid during pregnancy at a concentration of at least 3 mg / kg for 1-40 weeks;

[0042] A2) Oral administration of sialic acid during lactation, with a sialic acid concentration of at least 3 mg / kg or higher, for 1-104 weeks;

[0043] A3) The individual is orally administered sialic acid after birth, with the sialic acid concentration being at least 3 mg / kg or higher, for 1-104 weeks.

[0044] The promoting of height growth includes promoting an increase in bone length; preferably, the promoting of height growth includes promoting the growth of the tibia.

[0045] The promoting of brain-intestine co-development includes any one or more of the following B1)-B4):

[0046] B1) regulating the intestinal microenvironment; preferably, regulating the intestinal microenvironment includes reducing the abundance of harmful microorganisms and / or increasing the abundance of beneficial microorganisms; more preferably, the beneficial microorganisms include Bacteroides strains;

[0047] B2) promoting intestinal cell growth; preferably, at least further comprising promoting the proliferation of colon goblet cells;

[0048] B3) Improve cognitive and / or memory abilities;

[0049] B4) Improving neuronal plasticity; preferably, said improving neuronal plasticity comprises regulating the level of neural cell adhesion molecules.

[0050] The present invention has the following beneficial effects:

[0051] The present invention systematically studies the time period for oral sialic acid administration, the subject for oral sialic acid administration, the dosage of oral sialic acid administration, and the specific effects of oral sialic acid administration on gut-brain co-development.

[0052] The present invention discloses for the first time the effect of early supplementation of sialic acid on promoting early tibial growth and gut-brain co-development. Specifically, during the growth and development stage, sialic acid has a phased regulatory effect on height (especially tibia length) and weight. In behavioral experiments, sialic acid can significantly increase the time that offspring move in the central area of ​​the open field experiment, improve the discrimination and recognition index of the new object recognition experiment, and promote the occurrence of correct behavior in the Y maze experiment. In neurophysiology, sialic acid can increase the content of the nerve cell adhesion factor NCAM1 and improve neuronal plasticity. In terms of intestinal physiology, sialic acid can increase the number of colonic goblet cells, promote the proliferation of beneficial bacteria Bacteroides (Bacteroides), and increase the relative abundance of beneficial bacteria flora. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0054] Figure 1 This is a statistical diagram of the regulation of sialic acid on the early growth and development of young mice (A, body weight; B, bone length);

[0055] Figure 2 Statistical diagram of the effect of sialic acid on behavioral phenotypes (A, open field test, novel object recognition test at 3 weeks of age; B, Y maze test, novel object recognition test at 6 weeks of age);

[0056] Figure 3 This is a statistical diagram of the regulation of sialic acid on the level of neural cell adhesion molecule NCAM1;

[0057] Figure 4 This is a statistical diagram of the effect of sialic acid on intestinal flora;

[0058] Figure 5 Statistical graph of colon PAS staining results. DETAILED DESCRIPTION

[0059] Technical terms:

[0060] Neural cell adhesion molecule (NCAM) is a glycoprotein that mediates interactions between cells and between cells and the extracellular matrix. It plays a role in cell recognition and metastasis, tumor infiltration and growth, nerve regeneration, transmembrane signaling, learning and memory.

[0061] Neuronal plasticity: Neuronal plasticity refers to the ability of the nervous system to adapt to changes in its internal and external environment by altering its structure and function. This ability is reflected not only in the growth and connectivity of neurons, but also in changes in synaptic structure and function. These adaptive changes include structural plasticity and functional plasticity. For example, NCAM1 participates in synapse formation and neurite growth by regulating adhesion and connectivity between neurons. Neuronal plasticity plays a vital role in multiple aspects of the nervous system: through synaptic plasticity, neurons can form new connections or strengthen existing connections, thereby storing information and forming memories; after brain injury or disease, neuronal plasticity helps the nervous system reorganize and compensate for damaged functions, such as in recovery after stroke. Through neuronal plasticity, the nervous system continuously adapts to changes in the external environment, for example, in language acquisition and memory, skill learning, improved cognitive function and creativity, accelerated recovery from brain injury, enhanced emotional regulation and mental health, and delayed brain aging.

[0062] Bacteroides: Bacteroides are predominant bacteria in the human intestine and are believed to play a vital role in maintaining a healthy intestinal ecosystem. They are a candidate for next-generation probiotics. They help regulate the balance of the intestinal immune system and reduce inflammatory responses. They can enhance intestinal barrier function through interactions with host cells. Bacteroides homeostasis is crucial for intestinal health, and their metabolites (such as short-chain fatty acids) are beneficial to the host.

[0063] Goblet cells are mucus-secreting cells located between the columnar epithelial cells of the colonic mucosa. They are named for their large top-to-bottom shape, resembling a goblet. Goblet cells are single-cell glands whose primary function is to synthesize and secrete mucin, forming a mucosal barrier that protects epithelial cells.

[0064] In order to more clearly illustrate the overall concept of the present application, the following is a detailed description of the embodiments in conjunction with the accompanying drawings. In the following description, a large number of specific details are provided to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0065] Before further describing the specific embodiments of the present invention, it should be understood that the scope of the present invention is not limited to the specific embodiments described below. It should also be understood that the terminology used in the examples is intended to describe specific embodiments and is not intended to limit the scope of the present invention. The experimental procedures in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or according to the conditions recommended by the respective manufacturers.

[0066] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or combinations thereof.

[0067] When the embodiments provide numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the understanding of the prior art by those skilled in the art and the description of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention may also be used to implement the present invention.

[0068] The rats used for breeding experiments involved in the following examples are 10-week-old male and female SPF-grade (Specific pathogen free) Wistar, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. The ELISA kits in the following examples were purchased from Wuhan Yilai Ruite Biotechnology Co., Ltd. The determination method was in accordance with conventional analysis and there was no difference from other determinations. Animal behavioral experiments were carried out in a specific barrier, and the animal motion trajectory tracking system EthoVision XT was used for data acquisition and analysis. The equipment was purchased from Noldus Company in the Netherlands. The animal experiments met ethical requirements and were conducted strictly in accordance with ethical content. All experimental operations in animal experiments were carried out in specific animal barriers in accordance with the requirements of the animal room. The samples submitted for inspection were fixed with 4% paraformaldehyde. After the fixation was in good condition, they were trimmed, dehydrated, embedded, sliced, stained, and sealed in strict accordance with the pathological experimental detection procedures, and finally qualified samples were examined under a microscope. Among them, the reagents used in the present invention are all commonly used reagents, which can be purchased from conventional reagent production and sales companies.

[0069] In addition, the "water" mentioned in the present invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion exchange water, double distilled water, high-purity water, and purified water.

[0070] Example 1: Effects of different concentrations of sialic acid on bone length and body weight in young mice

[0071] In this example, the effects of different concentrations of sialic acid on the bone length and body weight of young mice were investigated. The specific experimental steps are as follows:

[0072] 1. Preparation of gavage solution

[0073] Different amounts of sialic acid were weighed and dissolved in saline to prepare a dose gradient of 0, 5, 10, and 50 mg / kg. Sialic acid (N-acetylneuraminic acid) was obtained from Bloomage Biosciences Co., Ltd., batch number 202112141.

[0074] 2. Experimental Animal Handling

[0075] Ten-week-old, SPF-qualified male and female Wistar rats were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. and housed in a standardized laboratory at 25±2°C, 50±5% relative humidity, and a 12-hour light / dark cycle. Experiments began after one week of acclimatization. The rats were housed in a 2:1 ratio of males to females, with three rats per cage, and allowed to mate freely for five days. After five days, the sexes were separated, and females were randomly re-assigned to two groups (solvent control group and sialic acid treatment group). During pregnancy and lactation, the rats were treated with either vehicle (normal saline) or 10 mg / kg sialic acid by gavage once daily between 9:00 AM and 12:00 PM. After birth, the offspring were further divided into four groups and treated with 0, 5, 10, or 50 mg / kg sialic acid by gavage once daily between 9:00 AM and 12:00 PM. The treatment period lasted from day 2 to 6 weeks of age. The specific groups are shown in Table 1.

[0076] Table 1

[0077]

[0078] 3. Experimental results measurement

[0079] The weight of the offspring was continuously measured and recorded during the feeding period. At the end of the experiment, the offspring were sacrificed and the right hind legs were uniformly removed when the animals were sacrificed for tissue collection. The length of the bones (tibia and femur) was measured and recorded. The experimental results are shown in Table 2 and Figure 1 shown.

[0080] Table 2

[0081]

[0082]

[0083] Note: There are two major variables in this experiment: whether the mother mice were sialic acid-intervened or not, and whether the offspring mice were sialic acid-intervened or not. In order to control the data to have only one variable each time, # and * are used to represent them respectively. Specifically, # can represent 10-0 group VS 0-0 group, 10-5 group VS 0-5 group, 10-10 group VS 0-10 group, 10-50 group VS 0-50 group. In this case, the dose of sialic acid intervention in offspring mice is controlled to be consistent, and the dose effect of sialic acid intervention in mother mice is studied; * can represent 0-5 group / 0-10 group / 0-50 group VS 0-0 group, or 10-5 group / 10-10 group / 10-50 group VS 10-0 group. In this case, the dose of sialic acid intervention in mother mice is controlled to be consistent, and the dose effect of sialic acid intervention in offspring mice is studied.

[0084] *, **, ***, **** represent that NANA supplementation had a significant effect on the weight and height of the offspring mice, P < 0.05, P < 0.01, P

[0085] <0.001, P<0.0001.

[0086] #, ## represent that supplementation of 10mg / kg NANA during pregnancy and lactation had a significant regulatory effect on the weight and height of offspring, P < 0.05,

[0087] P<0.01.

[0088] From Table 2 and Figure 1 visible.

[0089] Sialic acid intervention during pregnancy and lactation helped improve the physical development of offspring (#). At 3 weeks of age, the body weight of the 10-0 group was approximately 1.49 times that of the 0-0 group (78.48 / 52.6), and the tibia length of the 10-50 group was approximately 1.05 times that of the 0-50 group (2.68 / 2.543).

[0090] Postnatal sialic acid intervention also helps improve the physical development of pups (*). At 3 weeks of age, supplementation with NANA at 5 mg / kg or more significantly increased the body weight of pups. NANA supplementation had no significant effect on femur length. However, supplementation with NANA at 5 mg / kg or more significantly increased tibia length, with the 0-5 group approximately 1.27 times that of the 0-0 group (2.62 / 2.071). At 6 weeks of age, supplementation with NANA at 5 mg / kg or more significantly increased the body weight of pups. NANA supplementation had no significant effect on femur length, with the 10-50 group approximately 1.08 times that of the 10-0 group (3.33 / 3.09).

[0091] In general, the NANA dosage required to affect the growth indicators (weight, tibia) of offspring mice is relatively low, generally 5 mg / kg or above.

[0092] Example 2: Effects of different concentrations of sialic acid on cognition, memory-related behavior and related genes in young mice

[0093] In this example, the effects of different concentrations of sialic acid on cognition, memory-related behavior, and related genes in young mice were investigated. The specific steps are as follows:

[0094] 1. Experimental methods

[0095] The oral administration preparation and the feeding of experimental animals were the same as those in Example 1.

[0096] Behavioral experiments: Behavioral experiments were conducted at 3 and 6 weeks of age. The experimental protocol was based on the reference Cell Metabolism. 2021-03-01; 33(5): 923. Behavioral experiments were conducted using behavioral facilities in the animal room. ① Open field experiment: The rats were placed in a fixed position in the center of the bottom surface of the open field box, and video and timing were performed at the same time. When changing rats, the inner wall and bottom of the box were cleaned to remove any remaining biological information to avoid affecting the test results. The time the rats stayed in the center area and the number of times they entered the center area were recorded. ② New object recognition experiment: A test rat was placed in the field and allowed to explore freely. After habituation, two objects of similar size but different shapes and colors were placed in opposite corners of the box, and the test rats were allowed to explore the field with two objects. After an interval of 3 hours, the new object replaced one of the old objects, and the experiment was repeated. The time and number of times the rats explored the new and old objects were recorded. ③Y-maze experiment: The experimental animals are placed at the end of any arm of the Y-maze and allowed to explore freely. The camera system records the behavioral changes of the animals during this period and records the number of times they enter each arm.

[0097] The proportion of time spent in the central area in the open field test (%) was calculated as follows: the time the experimental animal spent in the central area / total test time*100%;

[0098] The Y-maze test score was calculated as: number of alternating turns / number of large turns;

[0099] The recognition index was calculated as: new object exploration time / (new object exploration time + old object exploration time).

[0100] 2. Experimental results:

[0101] The behavioral experimental results are as follows Figure 2 and shown in Table 3.

[0102] Table 3

[0103]

[0104] Note: There are two major variables in this experiment: whether the mother mice were sialic acid-intervened or not, and whether the offspring mice were sialic acid-intervened or not. In order to control the data to have only one variable each time, # and * are used to represent them respectively. Specifically, # can represent 10-0 group VS 0-0 group, 10-5 group VS 0-5 group, 10-10 group VS 0-10 group, 10-50 group VS 0-50 group. In this case, the dose of sialic acid intervention in offspring mice is controlled to be consistent, and the dose effect of sialic acid intervention in mother mice is studied; * can represent 0-5 group / 0-10 group / 0-50 group VS 0-0 group, or 10-5 group / 10-10 group / 10-50 group VS 10-0 group. In this case, the dose of sialic acid intervention in mother mice is controlled to be consistent, and the dose effect of sialic acid intervention in offspring mice is studied.

[0105] *, ** represent that NANA supplementation after birth significantly enhanced the cognitive and memory behaviors of offspring mice, P < 0.05, P < 0.01, respectively;

[0106] ## indicates that supplementation of 10 mg / kg NANA during pregnancy and lactation had a significant regulatory effect on the cognitive and memory behaviors of offspring mice, P < 0.01.

[0107] From Table 3 and Figure 2 visible.

[0108] Sialic acid intervention during pregnancy and lactation improved the cognitive and memory behaviors of offspring (#). At 3 weeks of age, the recognition index of the 10-50 group was approximately 1.68 times that of the 0-50 group (0.8222 / 0.4903).

[0109] Postnatal sialic acid intervention also helps improve cognitive and memory behaviors in offspring (*). At 3 weeks of age, postnatal NANA supplementation significantly increased the proportion of time spent in the center of the open field test and the recognition index in the novel object recognition test. The proportion of time spent in the center of the open field test in the 0-50 group was 2.26 times that of the 0-0 group (8.213 / 3.637), and the recognition index of the novel object recognition in the 10-50 group was 1.69 times that of the 10-0 group (0.8222 / 0.4866). At 6 weeks of age, postnatal NANA supplementation significantly increased the correct behavior score and recognition index in the Y-maze test. The Y-maze test score in the 0-50 group was 2.33 times that of the 0-0 group (0.7618 / 0.3274), and the recognition index of the novel object recognition in the 10-10 group was 1.31 times that of the 10-0 group (0.8316 / 0.6354).

[0110] Example 3: Effects of different concentrations of sialic acid on neuronal plasticity in young mice

[0111] In this example, the effects of different concentrations of sialic acid on neuronal plasticity in young mice were investigated. The specific steps were as follows:

[0112] 1. Experimental methods

[0113] The oral administration preparation and the feeding of experimental animals were the same as those in Example 1.

[0114] Physiological and biochemical experiments: Brain tissue was isolated and collected, and BCA working solution was prepared according to the instructions. The total protein concentration of the sample was detected, and the target indicator (NCAM1 level in the brain) was detected and analyzed using an ELISA kit.

[0115] 2. Experimental results:

[0116] The experimental results are as follows Figure 3 and as shown in Table 4.

[0117] Table 4

[0118]

[0119] Note: There are two major variables in this experiment: whether the mother mice were sialic acid-intervened or not, and whether the offspring mice were sialic acid-intervened or not. In order to control the data to have only one variable each time, # and * are used to represent them respectively. Specifically, # can represent 10-0 group VS 0-0 group, 10-5 group VS 0-5 group, 10-10 group VS 0-10 group, 10-50 group VS 0-50 group. In this case, the dose of sialic acid intervention in offspring mice is controlled to be consistent, and the dose effect of sialic acid intervention in mother mice is studied; * can represent 0-5 group / 0-10 group / 0-50 group VS 0-0 group, or 10-5 group / 10-10 group / 10-50 group VS 10-0 group. In this case, the dose of sialic acid intervention in mother mice is controlled to be consistent, and the dose effect of sialic acid intervention in offspring mice is studied.

[0120] *, ** represent that NANA supplementation after birth significantly enhanced neuronal plasticity in offspring mice, P < 0.05, P < 0.01, respectively;

[0121] # indicates that supplementation of 10 mg / kg NANA during pregnancy and lactation had a significant regulatory effect on neuronal plasticity in offspring mice, P < 0.05.

[0122] Depend on Figure 3 See Table 4.

[0123] Sialic acid treatment during pregnancy and lactation increased NCAM1 levels in the brains of offspring (#). At 3 weeks of age, NCAM1 levels in the 10-0 group were approximately 1.78 times higher than those in the 0-0 group (5.82 / 3.269). At 6 weeks of age, NCAM1 levels in the 10-0 group were approximately 2.13 times higher than those in the 0-0 group (3.504 / 1.645).

[0124] Postnatal sialic acid treatment also helped increase NCAM1 levels in offspring (*). At 3 weeks of age, NCAM1 levels in the 0-50 group were approximately 1.56 times higher than those in the 0-0 group (5.093 / 3.269). At 6 weeks of age, NCAM1 levels in the 0-50 group were approximately 3.25 times higher than those in the 0-0 group (5.352 / 1.645).

[0125] Example 4: Effects of different concentrations of sialic acid on the intestinal microenvironment of young mice

[0126] The specific steps are as follows:

[0127] 1. Experimental methods

[0128] The oral administration preparation and the feeding of experimental animals were the same as those in Example 1.

[0129] 16S rRNA amplicon sequencing and bioinformatics analysis of intestinal flora: Samples were collected, total genomes were extracted, and PCR amplification of the V3-V4 region of the 16S rDNA was performed. PCR products were recovered using agarose gels, and DNA libraries were constructed. The offline data were then spliced, annotated, and analyzed.

[0130] 2. Experimental results:

[0131] The relative abundance of the main different bacterial genera is as follows Figure 4 , as shown in Table 5, the PCoA analysis results are as follows Figure 4 shown.

[0132] Table 5 Relative abundance of main differential bacterial genera

[0133]

[0134]

[0135] Note: In this experiment, there are two major variables: whether the mother mice received sialic acid treatment or not, and whether the offspring mice received sialic acid treatment or not. To control for only one variable in each comparison, # and * are used to represent them, respectively. Specifically, # can represent the 10-0 group vs. the 0-0 group, the 10-5 group vs. the 0-5 group, the 10-10 group vs. the 0-10 group, and the 10-50 group vs. the 0-50 group. In this case, the sialic acid treatment dose of the offspring mice is controlled to be consistent, and the dose effect of sialic acid treatment of the mother mice is studied. * can represent the 0-5 group / 0-10 group / 0-50 group vs. the 0-0 group, or the 10-5 group / 10-10 group / 10-50 group vs. the 10-0 group. In this case, the sialic acid treatment dose of the mother mice is controlled to be consistent, and the dose effect of sialic acid treatment of the offspring mice is studied.

[0136] *Indicates that NANA supplementation after birth significantly enhanced the relative abundance of the main differential bacterial genera in offspring, P < 0.05;

[0137] ## indicates that supplementation of 10 mg / kg NANA during pregnancy and lactation had a significant regulatory effect on the relative abundance of the main differential bacterial genera in offspring mice, P

[0138] <0.01.

[0139] This example demonstrates that supplementation with NANA during pregnancy and lactation or after birth significantly affects the species β diversity of the intestinal flora of offspring. PCoA analysis at 3 weeks of age showed R = 0.5916, P = 0.001, and PCoA analysis at 6 weeks of age showed R = 0.3847, P = 0.001.

[0140] Supplementation of NANA during pregnancy and lactation significantly increased the abundance of beneficial bacteria Bacteroides in the offspring's intestines. At 3 weeks of age, the relative abundance of the 10-50 group was approximately 4.70 times that of the 0-50 group (6.729 / 1.432).

[0141] Postnatal NANA supplementation also significantly increased the abundance of beneficial bacteria, Bacteroides (*). At 3 weeks of age, the abundance of the 10-50 group was approximately 3.39 times that of the 10-0 group (6.729 / 1.987). At 6 weeks of age, the abundance of the 0-10 group was approximately 1.86 times that of the 0-0 group (0.5571 / 0.3).

[0142] Example 5: Effects of different concentrations of sialic acid on intestinal cells of young mice

[0143] In this example, the effects of different concentrations of sialic acid on intestinal cells of young mice were investigated. The specific steps are as follows:

[0144] 1. Experimental methods

[0145] The oral administration preparation and the feeding of experimental animals were the same as those in Example 1.

[0146] Colonic PAS staining was performed to count colonic goblet cells. The overall PAS staining procedure was as follows: dewaxing to desolvate, acidifying with periodic acid, rinsing with pure water, staining with Schiff's solution in the dark, developing with running water, and dehydrating and mounting. The specific steps were: dewaxing paraffin sections to desolvate, washing with distilled water, acidifying with periodic acid for 10 minutes, rinsing with pure water for 10 minutes, staining with Schiff's solution in the dark for 10 minutes, developing with running water for 5 minutes, and then rinsing with running water for 5 minutes; conventional dehydration, clearing, and mounting were performed. Images were acquired and analyzed using Image J software.

[0147] 2. Experimental results:

[0148] The results of colon PAS staining experiment are as follows Figure 5 and as shown in Table 6.

[0149] Table 6 Statistical results of PAS staining of colonic goblet cells

[0150]

[0151] Note: In this experiment, there are two major variables: whether the mother mice received sialic acid treatment or not, and whether the offspring mice received sialic acid treatment or not. To control for only one variable in each comparison, # and * are used to represent them, respectively. Specifically, # can represent the 10-0 group vs. the 0-0 group, the 10-5 group vs. the 0-5 group, the 10-10 group vs. the 0-10 group, and the 10-50 group vs. the 0-50 group. In this case, the sialic acid treatment dose of the offspring mice is controlled to be consistent, and the dose effect of sialic acid treatment of the mother mice is studied. * can represent the 0-5 group / 0-10 group / 0-50 group vs. the 0-0 group, or the 10-5 group / 10-10 group / 10-50 group vs. the 10-0 group. In this case, the sialic acid treatment dose of the mother mice is controlled to be consistent, and the dose effect of sialic acid treatment of the offspring mice is studied.

[0152] *, **, **** represent that NANA supplementation after birth has a significant enhancing effect on intestinal stem cells in offspring mice, P < 0.05, P < 0.01,

[0153] P < 0.0001;

[0154] #, ## represent that supplementation of 10mg / kg NANA during pregnancy and lactation had a significant regulatory effect on intestinal stem cells in offspring mice, P < 0.05,

[0155] P<0.01.

[0156] Depend on Figure 5 and as shown in Table 6.

[0157] Sialic acid intervention during pregnancy and lactation significantly affected the number of goblet cells in the intestines of offspring (#). At 3 weeks of age, the number of goblet cells in the 10-10 group was approximately 2.00 times that of the 0-10 group (452.7 / 226.8).

[0158] Postnatal NANA supplementation at 5 mg / kg or higher also significantly increased goblet cell numbers in offspring (*). At 3 weeks of age, the number of goblet cells in the 0-50 group was 2.37 times that of the 0-0 group (305.6 / 128.8), and the number in the 10-10 group was 2.35 times that of the 10-0 group (452.7 / 192.7). At 6 weeks of age, the number of goblet cells in the 0-10 group was 2.66 times that of the 0-0 group (544.8 / 204.8), and the number in the 10-10 group was 2.62 times that of the 10-0 group (819.3 / 312.8).

[0159] Example 6: Preparation of sialic acid solid beverage

[0160] Sialic acid can be used to prepare a solid beverage. The weight of the components in the solid beverage is as follows: 0.2-0.3 parts sialic acid, 5 parts blueberry powder, 15 parts oligofructose, 15 parts maltodextrin, and 0.1 parts zinc gluconate. Each component is prepared into a powder using conventional preparation methods. Each raw material powder is accurately weighed, thoroughly mixed, and packaged.

[0161] Example 7: Use of sialic acid in the preparation of oral solution

[0162] Sialic acid can be used to prepare an oral solution. The weight of the components in the oral solution is as follows: 0.2-0.3 parts of sialic acid, 10-20 parts of potassium sorbate, 20 parts of isomalto-oligosaccharide, 5 parts of citric acid, and 100 parts of purified water. The above components are prepared separately by weight, fully dissolved in purified water, filtered, sterilized, and canned to obtain the product.

[0163] Example 8: Sialic acid is used to prepare soft candies

[0164] Sialic acid can be used to prepare soft candies. The weight of the soft candies' components is as follows: 30 parts concentrated fruit juice, 0.2-0.3 parts sialic acid, 1 part fructo-oligosaccharide, 1 part isomalto-oligosaccharide, 0.5 parts citric acid, 40 parts white sugar, 1 part carrageenan, and 1.5 parts pectin. A mixed gel solution is prepared using conventional soft candy production methods, cast into shape, and packaged to obtain the finished product.

[0165] The foregoing is merely an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present application should all be included within the scope of the claims of the present application.

Claims

1. Application of sialic acid in the preparation of products for promoting height growth.

2. The use according to claim 1, characterized in that The promoting of height growth includes promoting an increase in bone length; preferably, the promoting of height growth includes promoting the growth of the tibia.

3. The use according to claim 1, characterized in that The application at least includes regulating the intestinal microenvironment; preferably, regulating the intestinal microenvironment includes reducing the abundance of harmful microorganisms and / or increasing the abundance of beneficial microorganisms; more preferably, the beneficial microorganisms include Bacteroides strains.

4. The use according to claim 1, characterized in that The application at least further comprises promoting the growth of intestinal cells; preferably, at least further comprises promoting the proliferation of colon goblet cells.

5. The use according to claim 1, characterized in that The use at least further comprises improving neuronal plasticity; preferably, improving neuronal plasticity comprises regulating the level of neural cell adhesion molecules.

6. A method for promoting height growth, characterized in that: Comprising administering sialic acid to the individual.

7. The method according to claim 6, characterized in that The concentration of sialic acid is at least 3 mg / kg or above; preferably, 5 mg / kg-500 mg / kg.

8. The method according to claim 6, characterized in that The method of administration includes oral administration.

9. The method according to claim 6, characterized in that The method of administration at least comprises taking a product containing sialic acid to the individual parent during pregnancy and / or lactation.

10. The method according to claim 6, characterized in that The method of administration at least comprises administering a product containing sialic acid to an individual after birth.

Images