Children formula food for improving intestinal inflammation and recovering key flora as well as preparation method and application of children formula food

Abstract

The invention provides a children formula food for improving intestinal inflammation and recovering key flora as well as a preparation method and application of the children formula food. The invention firstly provides the infant formula food, the infant formula food is infant formula food or children formula food, the infant formula food comprises 2 '-fucosyllactose and osteopontin, and the mass ratio of the 2'-fucosyllactose to the osteopontin is 1: (0.017-1.3). The children formula food disclosed by the invention can improve intestinal inflammation and recover key flora.

Description

Technical Field

[0001] The invention belongs to the technical field of pediatric formula food, and specifically relates to a pediatric formula food containing 2'-fucosyllactose and osteopontin, which is helpful to improve intestinal inflammation and restore key flora, and application thereof. Background Art

[0002] The first 1000 days of life is a window of opportunity for infant development. The dynamic development of the intestinal microecology complements the development of the intestinal immune system. The intestine is not only an important place for the digestion and absorption of nutrients, but also an important barrier to defend against external pathogens. Maintaining intestinal homeostasis and enhancing intestinal immune function are essential for maintaining body health. After birth, infants and young children are at high risk of gastrointestinal infections, inflammations and allergic diseases due to their immature digestive and immune functions and the unstable formation and development of intestinal flora. Breast milk is the first food that infants come into contact with. Breast milk is not only rich in nutrients, but also rich in immune active ingredients, growth factors and oligosaccharides. These ingredients in breast milk can not only provide immune support for infants and promote the maturation of intestinal barrier function, but also provide nutrients for probiotics such as bifidobacteria and lactobacilli, which helps to form a good microecological environment and ensure intestinal health. Literature results show that the infant's intestinal flora is closely related to the infant's development and the formation of the immune system. Establishing a healthy intestinal flora in the first 1000 days of life is important for the development and maturation of the intestinal mucosal immune system and the systemic immune system. The structure and relative abundance of the intestinal flora in early life will affect the probability of disease in the future.

[0003] In addition, there are a large number of keystone species in healthy humans, which play an important role in the species diversity and functional diversity of intestinal microorganisms and are indispensable. The specific composition of keystone microorganisms in infants and young children still needs to be further clarified. Some literature points out that the intestinal microbiome of infants is mainly composed of five phyla: Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria and Verrucomicrobia. A large number of studies have shown that the difference between the intestinal flora of diseased organisms and healthy organisms is mainly manifested in the decrease in the relative abundance of Firmicutes in the intestines of diseased organisms and the increase in the relative abundance of Proteobacteria.

[0004] Providing infants or children with a nutritional composition that can prevent and / or improve intestinal inflammatory symptoms and restore key flora is crucial for the healthy growth of infants and children. Summary of the invention

[0005] One object of the present invention is to provide a pediatric formula food that can improve intestinal inflammation and restore key flora.

[0006] Another object of the present invention is to provide a method for preparing the pediatric formula food.

[0007] Another object of the present invention is to provide related applications of the pediatric formula food.

[0008] On the one hand, the present invention provides a pediatric formula food, which is an infant formula food or a children's formula food. The pediatric formula food includes 2'-fucosyllactose and osteopontin, and the mass ratio of 2'-fucosyllactose to osteopontin is 1:(0.017 - 1.3).

[0009] 2'-fucosyllactose (2'-FL or 2-FL) is a trisaccharide structure formed by fucose and lactose and is a representative substance of fucosylated oligosaccharides. Commercially available this substance is usually prepared by microbial fermentation and has the same structure as the oligosaccharides found in human milk.

[0010] Osteopontin (OPN) is a secreted, phosphorylated integrin-binding protein and is an atypical immunomodulatory factor. Its role in regulating immune responses has been clearly established. OPN can promote cell survival and inhibit apoptosis, which plays a key role in intestinal mucosal protection. According to a specific embodiment of the present invention, in the pediatric formula food of the present invention, the raw material providing osteopontin can be a commercially available food-grade raw material with a high purity specifically for providing osteopontin, or it can be whey protein powder rich in osteopontin.

[0011] The present invention has found through research that combining 2'-fucosyllactose and osteopontin in a specific ratio has a synergistic effect on improving intestinal inflammatory symptoms and restoring key intestinal flora.

[0012] In the present invention, unless otherwise specified, the pediatric includes infants, toddlers or children. The infants are infants aged 0 - 12 months (including infants aged 0 - 6 months and older infants aged 6 - 12 months), the toddlers refer to toddlers aged 1 - 3 years, and the children refer to children aged 3 years to 18 years (under 19 years old), especially children at a relatively young age aged 3 years to 6 years (under 7 years old).

[0013] According to specific embodiments of the present invention, the pediatric formula food of the present invention is infant formula food (infant formula, toddler formula) or children's formula food (such as children's modified milk powder). Among them, in the infant formula food in terms of dry matter, the total protein content is 7.8 g - 22.8 g / 100 g (preferably, the proportion of whey protein in the total protein is 40% - 75%), the fat content is 14.9 g - 33.8 g / 100 g, and the carbohydrate content is 40.7 g - 79.3 g / 100 g. Among them, in the children's formula food in terms of dry matter, the total protein content is 16.5 g - 30 g / 100 g, the fat content is 10 g - 35 g / 100 g; the carbohydrate content is 40 g - 70 g / 100 g.

[0014] According to some specific embodiments of the present invention, the pediatric formula food of the present invention is infant formula food for special medical purposes.

[0015] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the mass ratio of 2'-fucosyllactose to osteopontin is 1:(0.1 - 1.3).

[0016] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the mass ratio of 2'-fucosyllactose to osteopontin is 1:(0.2 - 1.3).

[0017] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the mass ratio of 2'-fucosyllactose to osteopontin is 1:(0.25 - 1.3).

[0018] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.5 - 1.8 g / 100 g of food dry matter. The amount of osteopontin can be determined with reference to the aforementioned ratio range of 2'-fucosyllactose to osteopontin.

[0019] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.6 - 1.6 g / 100 g of food dry matter.

[0020] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.7 - 1.4 g / 100 g of food dry matter. The amount of osteopontin can be determined with reference to the aforementioned ratio range of 2'-fucosyllactose to osteopontin.

[0021] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.8 - 1.3 g / 100 g of food dry matter.

[0022] According to some specific embodiments of the present invention, the amount of 2'-fucosyllactose in the infant formula food of the present invention is 0.6 - 0.7 g / 100 g of food dry matter, 0.8 - 0.9 g / 100 g of food dry matter, 1.0 - 1.1 g / 100 g of food dry matter, 1.2 - 1.3 g / 100 g of food dry matter, 1.4 - 1.5 g / 100 g of food dry matter or 1.6 - 1.7 g / 100 g of food dry matter.

[0023] According to specific embodiments of the present invention, in the infant formula food of the present invention, the raw materials providing total protein include one or more of raw milk (cow / sheep), whole milk powder (cow / sheep), skim milk powder (cow / sheep), whey protein powder (cow / sheep), demineralized whey powder (cow / sheep), β-casein, etc.

[0024] According to specific embodiments of the present invention, in the infant formula food of the present invention, in addition to the basic raw materials containing milk fat, the raw materials providing fat may further include vegetable oil or OPO structured lipid.

[0025] According to specific embodiments of the present invention, for the infant formula food of the present invention, a part of the carbohydrates can come from basic raw materials containing lactose such as milk, whole milk powder and / or skim milk powder, etc. In addition, lactose raw materials can be additionally added to provide carbohydrates.

[0026] According to specific embodiments of the present invention, in the infant formula food of the present invention, in addition to the above components, it may further include conventional components of infant formula food or children's modified milk powder, for example, it may further include one or more of nutrients, dietary fiber, inositol, L-carnitine, docosahexaenoic acid, arachidonic acid, lutein, nucleotides.

[0027] In a specific embodiment, the infant formula food of the present invention is a dairy product, which can be a solid dairy product or a liquid dairy product. Specifically, it can be in the form of cheese, milk tablets, milk powder or liquid milk.

[0028] It can be understood that for different types of infant formula foods, the content of each functional substance in the final product should be appropriately adjusted within the range permitted by relevant standards and specifications.

[0029] In some specific embodiments of the present invention, the pediatric formula food of the present invention is milk powder, which can be prepared by adding / adjusting 2'-fucosyllactose and osteopontin to a basic formula powder. Among them, after the milk powder of the present invention is prepared, the contents of 2'-fucosyllactose and osteopontin can be determined by using conventional methods in the art, such as high performance liquid chromatography detection and analysis techniques. There is no particular limitation on the basic formula powder in the present invention, as long as it is a commonly used milk powder formula in the art. For example, the basic formula powder can be a common infant formula powder, toddler formula powder or children's milk powder. As a specific example, the basic formula powder of the present invention can use milk powder of animal origin as the main raw material, and can further be added with one or more of whey powder, vitamins, minerals, etc. As the milk powder of animal origin, cow milk powder or goat milk powder can be used.

[0030] According to some specific embodiments of the present invention, the pediatric formula food of the present invention has the following raw material composition:

[0031] Raw cow milk (partially skimmed available): 700 - 4700 parts (for example: 800 parts, 1000 parts, 3000 parts, etc.);

[0032] Whey powder: 0 - 600 parts (which can be converted into the corresponding amount of whey liquid, for example: 300 parts, 200 parts);

[0033] Whey protein powder and / or osteopontin raw material: 0 - 125 parts; preferably including osteopontin raw material: 1.5 - 22 parts; for example, whey protein powder: 0 - 100 parts (which can be converted into the corresponding amount of whey protein liquid, for example: 32 parts, 25 parts); highly pure raw material dedicated to providing osteopontin: 1.5 - 22 parts (for example: 8 parts, 10 parts);

[0034] Edible vegetable blended oil (may contain 1,3-dioleoyl-2-palmitoyl glycerol): 80 - 240 parts (for example: 230 parts, 180 parts);

[0035] Lactose: 0 - 550 parts (for example: 200 parts, 245 parts);

[0036] Minerals: 8 - 30 parts (for example: 18 parts, 15 parts);

[0037] Trace elements: 0.5 - 2 parts (for example: 1 part, 1.5 parts);

[0038] Vitamins: 2.5 - 10 parts (for example: 5 parts, 6 parts);

[0039] 2'-Fucosyllactose raw material: 6 - 17 parts (for example: 8.5 parts, 12.5 parts).

[0040] According to specific embodiments of the present invention, the raw material composition of the pediatric formula food of the present invention may further include one or more of α-lactalbumin (providing high-quality protein and osteopontin), linoleic acid (providing essential fatty acids), prebiotics (such as galactooligosaccharides, fructooligosaccharides, etc., providing support for intestinal health), probiotics, etc.

[0041] According to specific embodiments of the present invention, in the pediatric formula food of the present invention, each component raw material can be commercially sourced.

[0042] In the food of the present invention, the amounts of various substances should meet the requirements of relevant standards.

[0043] On the other hand, the present invention also provides a method for preparing the pediatric formula food described above, and the method includes:

[0044] Using a wet method, a dry method, or a combined wet and dry production process, the raw materials providing 2'-fucosyllactose and osteopontin are mixed with other raw materials in the formula. After the mixed material is filtered to remove impurities, it is homogenized and further used to prepare the pediatric formula food.

[0045] According to some specific embodiments of the present invention, in the method for preparing the pediatric formula food of the present invention, the homogenization temperature is not lower than 50°C, and the pressure is not lower than 120 bar.

[0046] According to some specific embodiments of the present invention, the method for preparing the pediatric formula food of the present invention further includes:

[0047] The homogenized material is concentrated until the dry matter weight content of the concentrate is 40% - 60%;

[0048] The concentrated material is sterilized at a temperature above 80°C for 10 - 60 seconds;

[0049] The sterilized material is dried through a spray drying tower (inlet air temperature 150°C - 200°C, outlet air temperature 80°C - 120°C, negative pressure -10 to -2 mmWG, high-pressure pump pressure 140 - 250 bar), and then introduced into a fluidized bed for secondary drying to obtain a powder.

[0050] In the preparation method of the present invention, ① according to the addition of 2'-fucosyllactose and osteopontin, the formula is optimized, which can improve intestinal inflammation and restore key flora, ensuring that infants and children can fully absorb and utilize these key nutrients. ② Mixing and formulation: The components are uniformly mixed in a specific ratio to ensure the consistency of the nutritional components in each formula food and avoid nutritional imbalance. Uniform mixing helps 2'-fucosyllactose and osteopontin play their best roles in the intestine, ensuring the consistency of the product's effects. ③ Production and processing: Advanced production processes such as homogenization and sterilization processes are adopted to ensure the safety and quality of the product, extend the shelf life, and at the same time retain the activity and stability of the nutritional components.

[0051] On the other hand, the present invention also provides the use of the pediatric formula food described above in the preparation of a product for improving intestinal inflammation symptoms and restoring key flora. The product can be the pediatric formula food of any of the foregoing embodiments of the present invention, or can be other products further prepared based on the pediatric formula food of the present invention.

[0052] According to a specific embodiment of the present invention, the improvement of intestinal inflammation symptoms and the restoration of key flora include one or more of the following:

[0053] Improving weight loss caused by intestinal inflammatory response;

[0054] Improving intestinal damage caused by inflammatory response;

[0055] Reducing the increase in serum myeloperoxidase (MPO) activity caused by intestinal inflammatory response;

[0056] Reducing the increase in serum diamine oxidase (DAO) level caused by intestinal inflammatory response;

[0057] Enhancing the level of intestinal goblet cells (improving the decrease in the level of circular cells caused by intestinal inflammatory response);

[0058] Improving intestinal barrier function;

[0059] Enhancing the intestinal physical and / or mucus barrier;

[0060] Increasing the expression level of intestinal mucin (improving the decrease in mucin level caused by intestinal inflammatory response);

[0061] Increasing the expression level of intestinal tight junction proteins (improving the decrease in tight junction protein level caused by intestinal inflammatory response);

[0062] Maintaining intestinal cell immune homeostasis;

[0063] Improving intestinal immune cell dysregulation caused by inflammatory response;

[0064] Enhancing the expression of intestinal CD4+ T cells;

[0065] Reduce the expression of intestinal CD8+ T cells;

[0066] Increase the ratio of intestinal CD4+ / CD8+ T cells;

[0067] Enhance the diversity and / or community richness of the intestinal microbiota;

[0068] Restore the intestinal keystone microbiota disorder or reduced diversity caused by inflammation or antibiotics;

[0069] Improve the reduction of intestinal microbial species caused by intestinal mucosal immune damage;

[0070] Increase the abundance of the intestinal genera Muribaculaceae and / or Lactobacillus;

[0071] Reduce the abundance of the intestinal genera Escherichia-Shigella and / or Alloprevotella.

[0072] Overall, the present invention mixes 2'-fucosyllactose and osteopontin in a certain proportion and uses them to prepare pediatric formula foods, which helps to improve intestinal inflammation and restore key microbiota, and is beneficial to the healthy growth of infants and children. Brief Description of the Drawings

[0073] Figure 1 Show the weight changes of rats in each group after LPS treatment for 6 h in a specific experiment of the present invention.

[0074] Figure 2 Show the weight loss of rats in each group after LPS treatment for 6 h in a specific experiment of the present invention.

[0075] Figure 3 Show the MPO activity in the serum of rats in each group in a specific experiment of the present invention.

[0076] Figure 4 Show the DAO level in the serum of rats in each group in a specific experiment of the present invention.

[0077] Figure 5 Show the effect on the number of goblet cells in the intestine of rats in each group in a specific experiment of the present invention.

[0078] Figures 6A - 6D Show the expression of mucin and tight junction proteins in the colon tissues of rats in each group in a specific experiment of the present invention, respectively.

[0079] Figures 7A - 7C Show the expression of immune cells in each group in a specific experiment of the present invention, respectively.

[0080] Figure 8 Show the effects of different groups on the number of OUTs in a specific experiment of the present invention.

[0081] Figure 9 Show the effects of different groups on the intestinal flora composition in a specific experiment of the present invention. Detailed implementation manners

[0082] Before further describing the specific implementation manners of the present invention, it should be understood that the protection scope of the present invention is not limited to the following specific implementation manners; it should also be understood that the terms used in the embodiments of the present invention are for describing specific implementation manners and not for limiting the protection scope of the present invention.

[0083] When an embodiment gives a numerical range, it should be understood that unless otherwise specified in the present invention, either endpoint of each numerical range and any value between the two endpoints can be selected.

[0084] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the art of this technology.

[0085] Except for the specific methods, devices, and materials used in the embodiments, according to the knowledge of those skilled in the art of this technology and the description of the present invention, any methods, devices, and materials of the prior art similar to or equivalent to those described in the embodiments of the present invention can also be used to implement the present invention.

[0086] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt the conventional techniques in this technical field.

[0087] In animal experiments, all reagent materials and test substances used were obtained through regular commercial purchases.

[0088] Efficacy experiment of 2'-fucosyllactose and osteopontin composition in improving intestinal inflammatory symptoms and restoring key flora

[0089] In this study, fecal samples were collected from healthy infants aged 3 - 6 months, and an infant microbiota humanized rat model was constructed using fecal transplantation technology. The rats were divided into 5 groups: a normal rat control group (NC group, gavaged with 1 mL of PBS daily), a model group (LPS group, gavaged with 1 mL of PBS daily), a 2’-FL group (gavaged with 1 mL of 2’-FL daily), an OPN group (gavaged with 1 mL of OPN daily), and a 2’-FL + OPN group (gavaged with 1 mL of a mixed solution of 2’-FL and OPN daily), with 12 rats in each group. After 14 days of gavage, the body weights of the rats in each group were measured. Except for the NC control group, the other 4 groups of rats were intervened with LPS (8 mg / kg body weight), and the body weights were measured again after continuous observation for 6 h. The gavage concentration of 2’-FL was 443 mg / kg body weight, the gavage concentration of OPN was 300 mg / kg body weight, and the dose of the mixed solution was the sum of the two. After the animal experiment, rat blood samples, as well as the terminal ileum and colon tissues, were collected. The activities of MPO and the contents of DAO in rat serum were detected using an ELISA kit (Solarbio, BC5715) according to the instructions.

[0090] The colon tissues were ground and homogenized in liquid nitrogen, and then total RNA was extracted from the tissues. The extracted RNA was reverse transcribed into cDNA using a reverse transcription kit (Qiagen, 51340). The expression levels of MUC2, ZO-1, Claudin-1, and Claudin-2 in the colon tissues were determined using a PCR instrument.

[0091] The expressions of CD4 and CD8 in the rat intestines were evaluated by immunofluorescence analysis, and the CD4 / CD8 ratio was calculated.

[0092] Finally, total DNA was extracted using a genomic DNA extraction kit, and sequencing was performed on an Illumina MiSeq high-throughput sequencer. The data of each sample were split from the downloaded data according to the Barcode sequence and the PCR amplification primer sequence. After removing the Barcode and primer sequences, the FLASH software was used for sequence splicing. The results related to the intestinal flora composition of each group were obtained.

[0093] The group information is shown in Table 1:

[0094] Table 1

[0095] Group Details NC Group Normal rat group, gavaged with PBS, no LPS treatment LPS Group Model animal group, gavaged with PBS, with LPS treatment 2’-FL Monomer intervention group, gavaged with 2’-FL, with LPS treatment OPN Monomer intervention group, gavaged with OPN, with LPS treatment 2’FL + OPN 2’FL + OPN, gavaged with the composition, with LPS treatment

[0096] Experimental results:

[0097] 1. Changes in the body weights of rats in each group before and after LPS treatment

[0098] The body weights of all experimental rats were measured before the start of LPS injection, and there were no significant differences among the groups. The results are as Figure 1As shown in the initial body weight part. After 6 h of LPS injection, the body weights of the rats in the injection groups (LPS group, 2’-FL group, OPN group, and 2’-FL+OPN group) were significantly decreased compared with those of the non-injected rats (NC group), and the results were as Figure 2 shown.

[0099] 2. Myeloperoxidase (MPO) activity in the serum of each group

[0100] Myeloperoxidase (MPO) in the serum is a leukocyte enzyme mainly secreted by activated macrophages, neutrophils, and monocytes. When the body is under oxidative stress, such as infection or fever, the body can release a large amount of MPO, generate oxygen free radicals, promote the oxidation of low-density lipoproteins in the body, and promote the occurrence and development of inflammatory reactions.

[0101] The results were as Figure 3 shown. Compared with the NC group, after LPS treatment, the activity of MPO in the serum of rats in the LPS group increased significantly (p<0.05), indicating that LPS treatment could cause an increase in the level of intestinal inflammation. Compared with the LPS group, the serum MPO activities in the 2‘-FL, OPN, and 2‘-FL+OPN groups were significantly decreased. Compared with the other two groups, the 2'-FL+OPN group could better reduce the activity of MPO in the serum, and there was no significant difference from the NC group. The composition showed a certain synergistic effect, indicating that the 2'-FL+OPN group could reduce the damage of inflammation to the intestine by reducing the MPO activity.

[0102] 3. Diamine oxidase (DAO) levels in the serum of each group

[0103] Diamine oxidase (DAO) is a structural enzyme specifically present in small intestinal mucosal epithelial cells. It is a key enzyme catalyzing the oxidation of histamine, and more than 90% of it exists in the small intestinal mucosa epithelium. The main role of DAO is to control the proliferation of intestinal mucosa, and its activity is closely related to the metabolism of intestinal mucosal epithelial cells. When the intestine is damaged or inflamed, the synthesis of DAO at the damaged site increases, and DAO then enters the blood. DAO exists stably in the serum, so it can reflect the situation of intestinal barrier damage and repair.

[0104] As Figure 4 shown, compared with the NC group, the serum DAO level of rats in the LPS group was significantly increased, indicating that the intestinal barrier of the rats was severely damaged; compared with the LPS model group, the serum DAO level of the rats in the intervention groups decreased significantly, indicating that their intestinal damage was protected to a certain extent. The 2’-FL+OPN composition was superior to the 2‘-FL and OPN monomers in reducing the increase in DAO caused by LPS treatment, and there was no significant difference from the NC group, showing a certain synergistic effect.

[0105] 4. Intestinal goblet cell level

[0106] The intestinal epithelium is mainly composed of four types of cells: absorptive cells, goblet cells, Paneth cells, and endocrine cells. These cells are constantly renewed to ensure the integrity of the intestinal epithelium. Some studies have shown that changes in the differentiation and secretory functions of goblet cells can lead to changes in the composition of the mucus layer, playing an important role in the occurrence and development of intestinal diseases.

[0107] The results were as Figure 5 shown. In the LPS group, there were epithelial cell shedding, inflammatory cell infiltration, and a decrease in goblet cells in the intestine. In the intervention groups, namely the 2’-FL group, OPN group, and 2’-FL + OPN group, the intestinal pathological conditions and the number of goblet cells were significantly improved. The number of goblet cells in the 2’-FL + OPN group was higher than that in the LPS group and the single-agent groups of 2‘-FL and OPN, indicating that 2'-FL and OPN have a certain effect on preventing tissue damage caused by LPS, and the combined effect of 2'-FL and OPN is better than the individual effects of 2'-FL and OPN.

[0108] 5. Gene expression levels of ileal mucin and tight junction proteins

[0109] Mucin is mainly composed of protein molecules, glycosyl compounds, amino acid residue modifiers, and metal ions, etc., and is one of the main components of the intestinal mucosal barrier. Members of the intestinal mucin family, such as mucin 1 (MUC1), mucin 2 (MUC2), mucin 5B (MUC5B), and mucin 13 (MUC13), are expressed in the intestine and play different functions. MUC2 consists of 5,100 amino acids and provides a protective barrier between the intestinal epithelial surface and the intestinal lumen. In addition to the mucosal barrier, tight junctions are composed of different occludin, claudin, and zonula occludens (ZOs). These three types of proteins and the actin cytoskeleton connect adjacent cells to form an epithelial barrier.

[0110] The mRNA expression levels of MUC2, Claudin-1, Claudin-2, and ZO-1 reflect the function of the intestinal barrier. The results were as Figures 6A - 6DAs shown, compared with the NC group, the mRNA expression level in the LPS group was significantly decreased (P<0.05); the groups of 2’-FL, OPN, and 2’-FL+OPN significantly changed this result and up-regulated the mRNA expression levels of MUC2, Claudin-1, Claudin-2, and ZO-1. In particular, in the 2'-FL+OPN group, the mRNA expression levels of MUC2, Claudin-1, Claudin-2, and ZO-1 were all significantly increased, showing the most obvious effect among all the intervention groups, and there was no significant difference compared with the NC group (Claudin-1), indicating a certain synergistic effect.

[0111] 6. Expression of immune cells in each group

[0112] See the expression of immune cells in each group Figures 7A - 7C . Compared with the NC group, the expression of CD4+ in the intestinal tract of rats in the LPS group was significantly decreased, and the expression of CD4+ T cells in rats in the groups of 2‘-FL, OPN, and 2’-FL+OPN was significantly increased. Compared with the NC group, the expression of CD8+ T cells in the LPS group was significantly increased, and the expression of CD8+ T cells in rats in the groups of 2‘-FL, OPN, and 2’-FL+OPN was significantly decreased. The result of CD4+ / CD8+T showed that compared with the NC control group, the ratio of CD4+ / CD8+ T cells in the LPS group was significantly decreased (P<0.05), indicating that the proportion of intestinal immune cells in rats was dysregulated and the immune function was damaged after LPS treatment. In the groups of 2‘-FL, OPN, and 2’-FL+OPN, the CD4+ / CD8+ of each group was significantly increased, especially in the 2'-FL+OPN group, the ratio of CD4+ / CD8+ T cells was increased most significantly, showing a certain coordinating effect.

[0113] 7. Effect of the combination of 2’-FL and OPN on intestinal flora

[0114] In this experiment, the effect of the 2’-FL+OPN composition on the intestinal flora of infants was further studied.

[0115] As Figure 8As shown, the specific OTUs in the NC group were significantly more than those in the LPS group, indicating that LPS treatment led to a significant reduction in the types of rat intestinal microorganisms. The number of OTUs in the 2'-FL+OPN group recovered, suggesting that the 2’-FL+OPN group could significantly improve the reduction in the types of intestinal microorganisms caused by intestinal mucosal immune injury. The results of α-diversity were shown in Table 1. The results showed that compared with the NC group, the observed species index, chao1 index, shannon index, and simpson index in the LPS group were all significantly decreased. Compared with the LPS group, the four indices in the 2'-FL+OPN group were all significantly increased and there was no significant difference from the NC group. The results indicated that 2'-FL+OPN could significantly increase the diversity and richness of the microbial community.

[0116] The results of α-diversity were shown in Table 2. The results showed that compared with the NC group, the observed species index, chao1 index, shannon index, and simpson index in the LPS group were all significantly decreased. Compared with the LPS group, the four indices in the 2'-FL+OPN group were all significantly increased. The results indicated that 2'-FL+OPN could significantly increase the diversity and richness of the microbial community.

[0117] Table 2. α-Diversity Analysis of Each Group

[0118] NC LPS 2'-FL + OPN chao1 618.98±48.23 472.70±15.60 621.86±10.93 observed_species 500.70±29.87 381.90±9.75 529.40±4.87 shannon 5.39±0.08 4.43±0.19 5.43±0.04 simpson 0.93±0.01 0.88±0.01 0.94±0.01

[0119] Using the 16s sequencing method, the flora composition of intestinal microorganisms was analyzed. The results of the flora composition analysis are shown in Figure 9The results showed that compared with the NC control group, the abundances of Lactobacillus and Muribaculaceae in the LPS group decreased significantly by 9.09% and 8.11% respectively, while the abundances of Escherichia-Shigella and Alloprevotella increased significantly by 7.15% and 11.14% respectively. In the 2'-FL + OPN group, Lactobacillus increased significantly by 10.12%, the abundance of Muribaculaceae increased significantly by 3.87%, the abundance of Alloprevotella decreased significantly by 21.11%, and the abundance of Escherichia-Shigella decreased by 2.01%. Changes in other genera included Eubacterium_coprostanoligenes_group, Romboutsia, Lachnoclostridium, Streptococcus, etc., but the changes were less significant or not significant. These changes indicated that the 2'-FL + OPN group successfully improved the intestinal flora structure compared with the LPS model group, especially reducing the abundances of some potentially harmful genera such as Escherichia-Shigella and Alloprevotella, and increasing the abundances of some potentially beneficial genera such as Lactobacillus and Muribaculaceae. The changes in other genera included some minor adjustments, making the flora structure of the 2'-FL + OPN group closer to that of the NC control group.

[0120] Example 1

[0121] This example provides a powdered infant formula for infants aged 0 - 6 months. The total protein content in this powdered formula is 10.5 g / 100 g, the 2'-fucosyllactose content is 0.6 - 0.7 g / 100 g, the osteopontin content is 0.15 - 0.175 g / 100 g, the fat content is 27.9 g / 100 g; and the carbohydrate content is 54.6 g / 100 g.

[0122] This example is achieved by compounding the following raw materials by weight. The raw material composition includes (preparing 1000 parts by weight):

[0123] 1000 parts of raw cow milk, 290 parts of demineralized whey powder, 230 parts of lactose, 226 parts of edible vegetable blended oil, 40.5 parts of galactooligosaccharide, 32 parts of whey protein powder (providing high-quality whey protein and osteopontin), 6.5 parts of 2'-fucosyllactose, 17.5 parts of fructooligosaccharide, 4 parts of phospholipid, 3.5 parts of compound vitamin, and 13 parts of mineral.

[0124] The infant formula powder of this embodiment is prepared by a dry-wet compound production process, which mainly includes: ingredient preparation, preheating, homogenization, concentration and sterilization, spray drying, and dry mixing to obtain the finished product. After testing, the various indicators of this finished product meet the requirements of the infant formula food standard.

[0125] Example 2

[0126] This embodiment provides a powdered formula food for older infants aged 6 to 12 months. The total protein content in this powdered formula food is 12 g / 100 g, 2'-fucosyllactose is 0.8 - 0.9 g / 100 g, osteopontin content is 0.2 - 0.225 g / 100 g, fat content is 23 g / 100 g; carbohydrate content is 57.6 g / 100 g.

[0127] This embodiment is achieved by compounding the following raw materials by weight. Its raw material composition includes (preparing 1000 parts by weight):

[0128] 1000 parts of raw cow milk, 275 parts of lactose, 225 parts of demineralized whey powder, 181 parts of edible vegetable blended oil, 130 parts of skim milk powder, 12.5 parts of galactooligosaccharide, 15 parts of whey protein powder (providing high-quality whey protein and osteopontin), 10 parts of casein, 5 parts of fructooligosaccharide, 4 parts of phospholipids, 3.5 parts of compound vitamins, 13 parts of minerals, 8.5 parts of 2'-fucosyllactose.

[0129] The infant formula powder of this embodiment is prepared by a dry-wet compound production process, which mainly includes: ingredient preparation, preheating, homogenization, concentration and sterilization, spray drying, and dry mixing to obtain the finished product. After testing, the various indicators of this finished product meet the requirements of the infant formula food standard.

[0130] Example 3

[0131] This embodiment provides a liquid formula milk for toddlers aged 12 to 36 months. The total protein content in this formula food, when converted to dry matter content, is 13.8 g / 100 g, 2'-fucosyllactose is 1.0 - 1.1 g / 100 g, osteopontin content is 0.25 - 0.275 / 100 g, fat content is 22 g / 100 g; carbohydrate content is 55.7 g / 100 g.

[0132] This embodiment is achieved by compounding the following raw materials by weight. Its raw material composition includes (preparing 1000 parts of dry matter):

[0133] 1000 parts of raw milk, 225 parts of skim milk powder, 220 parts of demineralized whey powder, 200 parts of lactose, 181 parts of edible vegetable blended oil, 13.5 parts of galactooligosaccharide, 9 parts of whey protein powder (providing high-quality whey protein and osteopontin), 6 parts of fructooligosaccharide, 1 part of casein, 4 parts of phospholipids, 3.5 parts of compound vitamins, 11 parts of minerals, 10.5 parts of 2'-fucosyllactose.

[0134] The infant formula food of this example is prepared by existing processes, mainly including: fresh milk - milk clarification - cold storage - mixing - high-temperature short-time sterilization - aseptic filling and sealing. Proteins, taurine and choline are added during the mixing process to obtain the finished product. After testing, the various indicators of this finished product meet the requirements of the infant formula food standards.

[0135] Example 4

[0136] This example provides a powdered children's formula milk powder. In this powdered formula food, the total protein content is 19 g / 100 g, the 2'-fucosyllactose is 1.2 - 1.3 g / 100 g, the osteopontin content is 0.3 - 0.325 g / 100 g, the fat content is 18 g / 100 g; the carbohydrate content is 55 g / 100 g.

[0137] This example is achieved by compounding the following raw materials by weight. Its raw material composition includes (preparing 1000 parts by weight):

[0138] 4400 parts of raw milk, 400 parts of demineralized whey powder, 25 parts of whey protein powder (providing high-quality whey protein and osteopontin), 15 parts of galactooligosaccharide, 15 parts of calcium carbonate, 7 parts of milk minerals, 6 parts of isomerized lactose, 3 parts of compound vitamins, 2 parts of phospholipids, 1 part of compound minerals, 1 part of choline, 12.5 parts of 2'-fucosyllactose.

[0139] The children's formula milk powder of this example is prepared by a dry-wet composite production process, mainly including: batching, preheating, homogenization, concentration and sterilization, spray drying, and dry mixing to obtain the finished product. After testing, the various indicators of this finished product meet the requirements of the children's formula milk powder food standards.

[0140] Example 5

[0141] This example provides a powdered children's formula milk powder. In this powdered formula food, the total protein content is 20 g / 100 g, the 2'-fucosyllactose is 1.4 - 1.5 g / 100 g, the osteopontin content is 0.35 - 0.375 g / 100 g, the fat content is 13.5 g / 100 g; the carbohydrate content is 58.5 g / 100 g;

[0142] This example is achieved by compounding the following raw materials by weight. Its raw material composition includes (preparing 1000 parts by weight):

[0143] 3,400 parts of raw cow milk, 275 parts of skim milk powder, 200 parts of lactose, 100 parts of demineralized whey powder, 12 parts of calcium carbonate, 5 parts of milk mineral salts, 2.5 parts of compound vitamins, 2 parts of phospholipids, 1 part of compound minerals, 1 part of choline, 14.5 parts of 2'-fucosyllactose.

[0144] The children's formulated milk powder of this example is prepared by a dry-wet compound production process, which mainly includes: batching, preheating, homogenization, concentration and sterilization, spray drying, and dry mixing to obtain the finished product. After testing, the various indicators of this finished product meet the requirements of the food standard for children's formulated milk powder.

[0145] Example 6

[0146] This example provides a powdered children's formulated milk powder. The total protein content in this powdered formula food is 20 g / 100 g, the content of 2'-fucosyllactose is 1.6 - 1.7 g / 100 g, the content of osteopontin is 0.4 - 0.425 g / 100 g, the fat content is 13.5 g / 100 g; the carbohydrate content is 58.5 g / 100 g;

[0147] This example is achieved by compounding the following raw materials in parts by weight. Its raw material composition includes (preparing 1,000 parts by weight):

[0148] 3,100 parts of raw cow milk, 360 parts of skim milk powder, 230 parts of lactose, 15 parts of galactooligosaccharide, 15 parts of calcium carbonate, 6 parts of isomerized lactose solution, 5 parts of milk mineral salts, 2.5 parts of compound vitamins, 2 parts of phospholipids, 1 part of compound minerals, 1 part of choline, 16.5 parts of 2'-fucosyllactose.

[0149] The children's formulated milk powder of this example is prepared by a dry-wet compound production process, which mainly includes: batching, preheating, homogenization, concentration and sterilization, spray drying, and dry mixing to obtain the finished product. After testing, the various indicators of this finished product meet the requirements of the food standard for children's formulated milk powder.

[0150] The above description of the examples is for the convenience of those of ordinary skill in the art to understand and use the invention. It is obvious that those skilled in the art can easily make various modifications to these examples and apply the general principles described herein to other examples without creative labor. Therefore, the present invention is not limited to the above examples, and the improvements and modifications made by those skilled in the art without departing from the scope of the present invention according to the disclosure of the present invention should be within the protection scope of the present invention.

Claims

1. A pediatric formula food, the pediatric formula food being an infant formula food or a children's formula food, the pediatric formula food comprising 2'-fucosyllactose and osteopontin, wherein the mass ratio of 2'-fucosyllactose to osteopontin is 1:(0.017-1.3); in, The infant formula food has a total protein content of 7.8g to 22.8g / 100g, a fat content of 14.9g to 33.8g / 100g, and a carbohydrate content of 40.7g to 79.3g / 100g, converted into dry matter; the children's formula food has a total protein content of 16.5g to 30g / 100g, a fat content of 10g to 35g / 100g, and a carbohydrate content of 40g to 70g / 100g, converted into dry matter.

2. The pediatric formula food according to claim 1, wherein: The mass ratio of 2'-fucosyllactose to osteopontin was 1:(0.1-1.3); Preferably, the mass ratio of 2'-fucosyllactose to osteopontin is 1:(0.2-1.3); Preferably, the mass ratio of 2'-fucosyllactose to osteopontin is 1:(0.25-1.3).

3. The pediatric formula food according to claim 1 or 2, wherein: The amount of 2'-fucosyllactose contained in the food is 0.5-1.8 g / 100 g of food dry matter.

4. The pediatric formula food according to claim 1 or 2, wherein: The amount of 2'-fucosyllactose in the food is 0.6-0.7g / 100g food dry matter, 0.8-0.9g / 100g food dry matter, 1.0-1.1g / 100g food dry matter, 1.2-1.3g / 100g food dry matter, 1.4-1.5g / 100g food dry matter or 1.6-1.7g / 100g food dry matter.

5. The pediatric formula food according to claim 1, wherein the raw materials thereof comprise: Raw milk: 700-4700 servings; Whey powder: 0-600 parts; Whey protein powder and / or osteopontin raw material: 0-125 parts; preferably including osteopontin raw material: 1.5-22 parts; Edible vegetable blended oil: 80-240 parts; Lactose: 0-550 parts; Minerals: 8-30 parts; Trace elements: 0.5-2 parts; Vitamins: 2.5-10 portions; 2'-fucosyllactose raw material: 6-17 parts.

6. The pediatric formula food according to any one of claims 1 to 5, which is milk powder or liquid milk.

7. A method for preparing the pediatric formula food according to any one of claims 1 to 6, the method comprising: The raw materials providing 2'-fucosyllactose and osteopontin are mixed with other raw materials in the formula by adopting a wet method, a dry method or a dry-wet composite production process. The mixed materials are homogenized after filtering and removing impurities, and are further used to prepare the pediatric formula food.

8. The preparation method according to claim 7, further comprising: The homogenized material is concentrated to a dry matter weight content of 40% to 60%; The concentrated material is sterilized at above 80°C for 10 to 60 seconds; The sterilized material is dried by a spray drying tower (inlet air temperature 150°C ~ 200°C, exhaust air temperature 80°C ~ 120°C, negative pressure -10 ~ -2mmWG, high pressure pump pressure 140 ~ 250bar), and then introduced into a fluidized bed for secondary drying to obtain a powder.

9. Use of the pediatric formula food according to any one of claims 1 to 6 in the preparation of a product for improving intestinal inflammatory symptoms and restoring key flora.

10. The use according to claim 9, wherein: Improving intestinal inflammation symptoms and restoring key flora include one or more of the following: Improve weight loss caused by intestinal inflammatory response; Improve intestinal damage caused by inflammatory response; Reduce the increase in serum myeloperoxidase (MPO) activity caused by intestinal inflammatory response; Reduce the increase in serum diamine oxidase (DAO) levels caused by intestinal inflammatory response; Improve the level of intestinal goblet cells (improve the decrease in the level of ring cells caused by intestinal inflammatory response); Improve intestinal barrier function; strengthening the intestinal physical and / or mucus barrier; Increase the expression level of intestinal mucin (improve the decrease of mucin level caused by intestinal inflammatory response); Increase the expression level of intestinal tight junction proteins (improve the decrease in tight junction protein levels caused by intestinal inflammatory response); Maintain intestinal cell immune homeostasis; Improve intestinal immune cell disorders caused by inflammatory response; Increase intestinal CD4+T cell expression; Reduce intestinal CD8+T cell expression; Improve the ratio of intestinal CD4+ / CD8+T cells; Improving the diversity and / or richness of the intestinal microbiome; Restoring the disturbance or reduced diversity of the intestinal cornerstone flora caused by inflammation or antibiotics; Improve the reduction of intestinal microbial species caused by intestinal mucosal immune damage; Increase the abundance of the enteric bacterial genera Muribaculaceae and / or Lactobacillus; Reduction of the abundance of intestinal Escherichia-Shigella and / or Alloprevotella.

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